Submitted Successfully!
To reward your contribution, here is a gift for you: A free trial for our video production service.
Thank you for your contribution! You can also upload a video entry or images related to this topic.
Version Summary Created by Modification Content Size Created at Operation
1
Ivana Šola
 + 5934 word(s) 5934 2022-01-21 07:25:26 |
2 format correct
Vivi Li
 + 14 word(s) 5948 2022-01-21 09:47:03 |

Video Upload Options

We provide professional Video Production Services to translate complex research into visually appealing presentations. Would you like to try it?
No, upload directly Yes

Confirm

Are you sure to Delete?
Yes No
Cite
If you have any further questions, please contact Encyclopedia Editorial Office.
Šola, I. Biopotential of Underutilized Rosaceae Inflorescences. Encyclopedia. Available online: https://encyclopedia.pub/entry/18599 (accessed on 20 November 2024).
Šola I. Biopotential of Underutilized Rosaceae Inflorescences. Encyclopedia. Available at: https://encyclopedia.pub/entry/18599. Accessed November 20, 2024.
Šola, Ivana. "Biopotential of Underutilized Rosaceae Inflorescences" Encyclopedia, https://encyclopedia.pub/entry/18599 (accessed November 20, 2024).
Šola, I. (2022, January 21). Biopotential of Underutilized Rosaceae Inflorescences. In Encyclopedia. https://encyclopedia.pub/entry/18599
Šola, Ivana. "Biopotential of Underutilized Rosaceae Inflorescences." Encyclopedia. Web. 21 January, 2022.
Biopotential of Underutilized Rosaceae Inflorescences
Edit
This entry is adapted from the peer-reviewed paper 10.3390/plants11030271

This work aims to assess the biopotential of the young inflorescence tissues of Prunus, Malus and Chaenomeles in order to evaluate the possibility of their application in the food industry, and to provide a polyphenolic fingerprint for their quality control. The contents of different bioactive compounds and their antioxidant capacities were spectrophotometrically measured, the main phenolic compounds were identified and quantified using LC-DAD-MS, the antidiabetic potential was determined using α-amylase and α-glucosidase inhibition assays, the anti-inflammatory potential was determined using a 5-lipoxygenase inhibition assay, and the cytotoxicity was determined by MTT assay. Using one-way ANOVA, principal component analysis, hierarchical clustering and Pearson’s correlation coefficient, the relations between the samples, and between the samples and the measured parameters, were revealed. In total, 77 compounds were identified. The concentration of sugars was low in M. purpurea, at 1.56 ± 0.08 mg/g DW. The most effective sample in the inhibition of antidiabetic enzymes and anti-inflammatory 5-lipoxygenase was C. japonica. The inhibition of α-glucosidase was strongly positively correlated with the total and condensed tannins, procyanidin dimers and procyanidin tetramer, and was very strongly correlated with chlorogenic acid. In α-amylase inhibition, C. japonica and P. serrulata ‘Kiku Shidare Zakura’ were equally efficient to the standard inhibitor, maltose. The most effective in the growth and proliferation inhibition of HepG2, HCT116 and HaCaT cells was P. avium. The results suggest Prunus, Malus and Chaenomeles inflorescences as functional food ingredients.

5-lipoxygenase α-amylase α-glucosidase Chaenomeles colorectal carcinoma hepatocellular carcinoma keratinocytes Malus metabolomics Prunus

1. Introduction

Inflorescence, as a precursor of a fruit, is composed of several types of metabolically very active tissues. In recent years, the biopotential of the bioactive compounds of this plant structure has been recognized. Namely, the significant bioactivities of inflorescences of industrial hemp [1], Musa species [2], Sorbus aucuparia [3], Cistus salviifolius [4], Lonicera japonica [5], and Caryota urens [6] have been revealed. Moreover, the flower buds of C. salviifolius have been shown to have a higher concentration of phenolics, better antioxidant and anti-superoxide dismutase activity, and stronger cytotoxic activity against human breast cancer and ovarian cancer cells than leaves [4]. Recently, the intrinsic antiradical activity of industrial hemp’s inflorescences’ water extracts and mechanisms related to their anti-inflammatory, antiproliferative and antimycotic activity have been revealed [1]. The inflorescences of Musa species are one of the most widely consumed vegetables in the Southeast Asian region, and their biological activities are well known.
The use of inflorescences of Rosaceae varieties, except those of Sorbus spp. [3], is not that common. Sorbus spp. inflorescences have a higher concentration of total phenolics than the commonly consumed fruits [3]. Furthermore, the inflorescences of different Sorbus species have higher antioxidant activity than their leaves or fruits [7]. Moreover, the extremely high content of phenolics in S. aucuparia inflorescences suggests their great potential as a source for natural health products [3][7]. Different organs of Prunus, Malus, and Chaenomeles taxa have already been investigated for their phytochemical, nutraceutical and bioactivity potential [8][9][10][11][12][13]. However, the inflorescences have been neglected so far.
Therefore, the aim of this work was to assess the biopotential of the young inflorescence tissues of selected Prunus, Malus, and Chaenomeles using a combination of spectrometric, chromatographic, cell culture and chemometric analyses, as well as to evaluate the possibility of their application in the food industry. For that purpose, we (a) spectrophotometrically measured the content of different types of bioactive compounds (total phenolics, flavonoids, non-flavonoids, total and condensed tannins, and soluble sugars), and their antioxidant capacity by the three methods (ABTS, FRAP and DPPH); (b) identified and quantified the main phenolic compounds using the LC-DAD-MS method; (c) determined the antidiabetic potential of the inflorescences using α-amylase and α-glucosidase inhibition assays; (d) measured the anti-inflammatory potential using a 5-lipoxygenase inhibition assay; (e) assessed the cytotoxicity toward HepG2, HCT 116 and HaCaT cells by MTT assay; and (f) statistically—using one-way ANOVA, principal component analysis, hierarchical clustering and Pearson’s correlation coefficient—revealed the relations between the samples, as well as between the samples and the measured parameters. This study is the first to present a comprehensive LC-DAD-MS polyphenolic profile of the inflorescences of Prunus avium (L.) L. (Pa), Prunus serrulata Lindl. (Ps), Prunus serrulata ‘Kiku Shidare Zakura’ (PsKss), Malus x purpurea (E.Barbier) Ehder (Mp), Malus floribunda Siebold ex Van Houtte (Mf) and Chaenomeles japonica (Thunb.) Lindl. ex Spach (Cj), providing a fingerprint for the future quality control of these biomaterials, and the first report on their bioactivity.

2. Spectrophotometric Analysis of the Phytochemical Content and Antioxidant Capacity of the Inflorescences

All of the the tested samples had a similar or higher amount of total phenolics than the commonly used flower buds of Capparis spinosa [14]. The highest amount of total phenolics (53.12 ± 0.79 mg GAE/g DW), flavonoids (38.89 ± 4.04 mg CE/g DW) and nonflavonoids (31.32 ± 0.71 mg GAE/g DW) were recorded in PsKss (Table 1). Because only about 10% of the medicinal plant material has total phenolics in a concentration higher than 5% DW of GAE [7][15], PsKss inflorescence is among the plant materials with the highest concentration of these compounds. Compared to the concentration of the total flavonoids in many other commonly used medicinal plants [15], the tested inflorescences had a similar or higher concentration, suggesting their potential for the food industry. Moreover, very recently, it was shown that inflorescences of Pa have far more total phenolics and flavonoids than stems and kernels [16].
Table 1. Concentration (in mg/g dry weight) of the total phenolics (TP), total flavonoids (TF), total nonflavonoids (TNF), total tannins (TT), condensed tannins (CT) and soluble sugars (SS), and the antioxidant capacity (ABTS, FRAP and DPPH) of Rosaceae inflorescences.
  Prunus avium Prunus serrulata Prunus serrulata ‘Kiku Shidare Zakura’ Prunus yedoensis Malus purpurea Malus floribunda Chaenomeles japonica
TP (mg GAE/g DW) 27.83 ± 0.69 f 46.84 ± 0.75 b 53.12 ± 0.79 a 37.31 ± 0.48 e 40.29 ± 0.64 c 46.74 ± 0.93 b 39.42 ± 0.97 d
TF (mg CE/g DW) 13.57 ± 0.84 f 32.35 ± 2.13 b 38.89 ± 4.04 a 25.78 ± 1.03 d 13.43 ± 0.82 f 23.83 ± 0.88 e 29.45 ± 0.65 c
TNF (mg GAE/g DW) 16.40 ± 0.91 g 29.35 ± 0.83 b 31.32 ± 0.71 a 22.21 ± 0.86 e 23.52 ± 0.56 d 28.85 ± 0.58 c 18.73 ± 0.48 f
TT (mg CE/g DW) 27.26 ± 0.22 g 59.44 ± 1.32 f 71.59 ± 0.33 d 83.55 ± 0.55 b 64.32 ± 0.45 e 107.85 ± 1.09 a 80.27 ± 0.33 c
CT (mg CE/g DW) 4.25 ± 0.33 e 7.74 ± 1.02 d 6.99 ± 0.17 d 16.52 ± 0.01 b 15.45 ± 0.10 b 10.98 ± 0.24 c 51.68 ± 0.38 a
SS (mg SE/g DW) 3.37 ± 0.06 c 3.04 ± 0.05 d 2.41 ± 0.10 e 8.61 ± 0.12 a 1.56 ± 0.08 f 3.58 ± 0.06 b 3.27 ± 0.07 c
ABTS (mg TE/g DW) 22.86 ± 4.61 e 49.41 ± 7.23 b 61.32 ± 5.84 a 36.63 ± 4.32 c 28.78 ± 2.52 d 47.78 ± 6.26 b 35.05 ± 4.25 c
FRAP (mg TE/g DW) 27.89 ± 0.60 g 51.68 ± 0.12 b 58.06 ± 0.78 a 40.28 ± 1.18 d 29.12 ± 0.83 f 44.36 ± 0.85 c 36.36 ± 1.50 e
DPPH (mg TE/g DW) 25.47 ± 2.57 d 52.95 ± 4.22 b 69.42 ± 3.27 a 39.21 ± 4.86 c 25.10 ± 3.25 d 40.38 ± 3.4 c 39.61 ± 3.95 c
Values represent the mean ± standard deviation of three replicates. Different letters indicate a significant difference among the values in a row (ANOVA, Duncan test, p ≤ 0.05). GAE = gallic acid equivalent, CE = catechin equivalent, SE = sucrose equivalent, TE = trolox equivalent.
In order to reduce the global rise of overweightness and obesity, the content of sugars, as a source of calories, in food should be diminished. As expected, the level of sugars in the inflorescences was lower than that in the more often-used fruits. The interval of the concentrations in the inflorescences was between 1.56 ± 0.08 mg/g DW in Mp and 8.61 ± 0.12 mg/g DW in Py (Table 1), which is relatively low compared to the concentrations recorded in M. domestica fruits [17] and Pa fruits [10]. Namely, the ripe flesh and skin of cultivated stone and pome fruits usually contain soluble sugars in the amount of 70–90% of the DW [18]. This is one of the main reasons why Rosaceae inflorescences have a high potential in the food industry, and should be considered a potent biomaterial.
The antioxidant potential was assessed using three standard methods (ABTS, FRAP and DPPH), and each of them revealed PsKss as the most potent sample (Table 1). Because this taxon had significantly more total phenolics, flavonoids, nonflavonoids (Table 1), and total identified hydroxycinnamic acids than the other samples (Table 2), with the predominant ones being caffeic acid hexoside 1,3-caffeoylquinic acid, 4-caffeoylquinic acid, 5-caffeoylquinic acid 1 (chlorogenic acid), and p-coumaric acid hexoside 1 and 2, we assume that these compounds significantly contributed to its antioxidant potential. The same relations between the content of phenolic compounds and antioxidant activity between different parts of Pa have already been revealed as well [16][19]. The antioxidant activity of Cj inflorescences was similar to that of its fruits [20]. However, inflorescences of Pa showed higher antioxidant potential than the stems and kernels [16]. Mf had a higher antioxidant capacity than Mp (Table 1), and this is probably related to the higher amount of identified hydroxycinnamic acids, especially chlorogenic acid, flavanols, flavanones, flavonols and procyanidins (Table 2). Both Mf and Mp dry inflorescences showed higher antioxidant activity than the fresh flowers of different Malus cultivars [21], highlighting their potential for use in the food industry.
Table 2. Concentration (mg/g DW ± SD) of the individual phenolic compounds in Rosaceae inflorescences.
    Prunus avium Prunus serrulata Prunus serrulata ‘Kiku Shidare Zakura’ Prunus yedoensis Malus purpurea Malus floribunda Chaenomeles japonica
1 Gallic acid 0.28 ± 0.06 b 0.15 ± 0.04 c 0.06 ± 0.01 d 0.38 ± 0.03 a 0.17 ± 0.04 c 0.03 ± 0.01 d nd
  Total identified
hydroxybenzoic acids		 0.28 ± 0.06 b 0.15 ± 0.04 c 0.06 ± 0.01 d 0.38 ± 0.03 a 0.17 ± 0.04 c 0.03 ± 0.01 d nd
2 Caffeic acid 1.95 ± 0.27 c 1.25 ± 0.14 c 2.50 ± 0.08 b 3.90 ± 0.10 a nd nd 0.53 ± 0.08 d
3 Caffeic acid hexoside 1 0.14 ± 0.03 b 0.39 ± 0.06 b 15.48 ± 2.23 a 0.10 ± 0.01 b 0.05 ± 0.01 b 0.23 ± 0.02 b 0.75 ± 0.04 b
4 Caffeic acid hexoside 2 nd nd nd 5.62 ± 0.25 a nd nd 0.02 ± 0.00 b
5 Caffeic acid dihexoside 0.29 ± 0.01 b 0.19 ± 0.04 c 0.41 ± 0.02 a nd nd nd nd
6 3-caffeoylquinic acid nd 0.36 ± 0.04 b 1.70 ± 0.25 a 0.54 ± 0.03 b nd nd 0.02 ± 0.00 c
7 4-caffeoylquinic acid nd 0.27 ± 0.08 a 0.35 ± 0.02 a nd 0.13 ± 0.06 b 0.15 ± 0.02 b 0.07 ± 0.01 b
8 5-caffeoylquinic acid 1 1.59 ± 0.18 d 0.59 ± 0.17 e 0.47 ± 0.02 e 5.75 ± 0.15 c 1.41 ± 0.30 d 8.41 ± 0.69 a 7.04 ± 0.23 b
9 5-caffeoylquinic acid 2 0.43 ± 0.36 a nd nd 0.27 ± 0.01 a 0.31 ± 0.13 a nd 0.14 ± 0.02 a
10 di-caffeoylquinic acid 1 3.12 ± 0.08 b 0.15 ± 0.03 c 0.31 ± 0.02 c 7.06 ± 0.82 a 0.65 ± 0.14 c 0.50 ± 0.03 c 2.83 ± 0.30 b
11 di-caffeoylquinic acid 2 0.13 ± 0.01 b nd nd 0.29 ± 0.01 b 0.16 ± 0.06 b 1.02 ± 0.15 a nd
12 di-caffeoylquinic acid 3 0.17 ± 0.03 a nd nd nd nd nd nd
13 3-feruloylquinic acid 0.24 ± 0.04 a 0.03 ± 0.00 c 0.05 ± 0.00 bc 0.08 ± 0.00 b nd nd 0.004 ± 0.001 c
14 5-feruloylquinic acid 0.26 ± 0.03 a 0.06 ± 0.01 d 0.11 ± 0.00 bc 0.09 ± 0.01 cd 0.01 ± 0.00 e 0.13 ± 0.04 b 0.01 ± 0.00 e
15 3-p-coumaroylquinic acid 0.52 ± 0.08 b 0.14 ± 0.02 d 0.21 ± 0.01 c 0.73 ± 0.03 a 0.03 ± 0.01 e 0.15 ± 0.01 cd 0.001 ± 0.000 e
16 4-p-coumaroylquinic acid nd 0.27 ± 0.08 a 0.35 ± 0.02 a nd 0.13 ± 0.06 b 0.15 ± 0.02 b 0.07 ± 0.02 b
17 5-p-coumaroylquinic acid 1 0.39 ± 0.02 a 0.09 ± 0.01 d 0.07 ± 0.00 d 0.33 ± 0.03 b 0.10 ± 0.01 d 0.19 ± 0.03 c 0.12 ± 0.03 d
18 5-p-coumaroylquinic acid 2 0.03 ± 0.00 c 0.09 ± 0.01 b 0.08 ± 0.01 bc 0.10 ± 0.01 b 0.07 ± 0.06 bc 0.06 ± 0.02 bc 0.25 ± 0.04 a
19 p-coumaric acid hexoside 1 0.23 ± 0.06 b 0.21 ± 0.04 b 0.31 ± 0.01 a 0.09 ± 0.00 c 0.04 ± 0.01 cd 0.22 ± 0.04 b 0.001 ± 0.000 d
20 p-coumaric acid hexoside 2 0.37 ± 0.06 b 1.42 ± 0.15 a 1.34 ± 0.05 a 0.21 ± 0.01 c 0.02 ± 0.00 d 0.11 ± 0.01 cd 0.13 ± 0.00 cd
  Total identified
hydroxycinnamic acids		 9.87 ± 0.08 b 5.51 ± 0.06 c 23.70 ± 0.18 a 25.16 ± 0.10 a 3.12 ± 0.06 d 11.31 ± 0.09 b 11.93 ± 0.05 b
21 Catechin 0.23 ± 0.04 e 2.61 ± 0.27 a 2.46 ± 0.09 a 0.91 ± 0.02 d nd 1.99 ± 0.16 b 1.67 ± 0.06 c
22 Epicatechin 1.10 ± 0.15 d 0.91 ± 0.10 d 1.81 ± 0.06 b 6.92 ± 0.18 a 1.18 ± 0.18 d 1.38 ± 0.22 c 0.40 ± 0.06 e
  Total identified flavanols 1.33 ± 0.09 e 3.52 ± 0.18 cd 4.27 ± 0.07 b 7.83 ± 0.10 a 1.18 ± 0.18 e 3.38 ± 0.19 bc 2.08 ± 0.06 de
23 Eriodictyol hexoside 1 0.03 ± 0.00 b nd nd nd 0.63 ± 0.04 b 2.92 ± 0.86 a nd
24 Eriodictyol hexoside 2 nd nd nd nd 0.35 ± 0.09 b 1.02 ± 0.15 a nd
25 Naringenin hexoside nd nd nd nd nd nd 0.62 ± 0.04 a
  Total identified flavanones 0.03 ± 0.00 b nd nd nd 0.98 ± 0.06 b 3.93 ± 0.50 a 0.62 ± 0.04 b
26 Quercetin-glycoside 0.18 ± 0.01 c 0.13 ± 0.01 d 0.21 ± 0.02 b 0.36 ± 0.02 a nd nd nd
27 Quercetin-3-rutinoside 2.78 ± 0.28 b 0.58 ± 0.10 d 4.79 ± 0.13 a 1.93 ± 0.03 c 0.08 ± 0.03 e 0.21 ± 0.03 e 0.72 ± 0.04 d
28 Quercetin-3-rhamnoside hexoside nd nd nd nd 0.58 ± 0.21 a 0.41 ± 0.06 a nd
29 Quercetin-hexoside pentoside nd 0.25 ± 0.08 b 0.59 ± 0.01 a nd nd nd nd
30 Quercetin-rhamnoside dihexoside 1 0.15 ± 0.01 b nd nd 0.20 ± 0.00 a nd nd nd
31 Quercetin-rhamnoside dihexoside 2 nd 0.02 ± 0.01 c 0.05 ± 0.00 b 0.47 ± 0.01 a nd nd nd
32 Quercetin-3-galactoside 0.26 ± 0.02 c nd nd 0.98 ± 0.01 b 1.21 ± 0.27 a 0.309 ± 0.070 c 0.17 ± 0.01 c
33 Quercetin-3-glucoside 0.03 ± 0.00 d 0.19 ± 0.02 a 0.12 ± 0.01 b nd 0.02 ± 0.00 d 0.089 ± 0.004 c 0.02 ± 0.00 d
34 Quercetin-3-rhamnoside nd nd nd nd 2.26 ± 0.38 b 4.31 ± 0.51 a nd
35 Quercetin-3-xyloside 0.01 ± 0.00 e 0.02 ± 0.00 de 0.06 ± 0.00 c 0.56 ± 0.04 a 0.09 ± 0.01 b 0.05 ± 0.01 cd 0.02 ± 0.00 de
36 Quercetin-arabinofuranoside 0.13 ± 0.04 c 0.15 ± 0.02 c 0.25 ± 0.03 b 0.17 ± 0.00 c 0.03 ± 0.01 d 0.31 ± 0.04 a 0.004 ± 0.000 d
37 Quercetin-arabinopyranoside 0.01 ± 0.00 c nd nd nd 2.07 ± 0.20 a 1.61 ± 0.05 b nd
38 Quercetin-acetyl hexoside 1 nd 4.00 ± 0.40 a 1.53 ± 0.07 b 0.22 ± 0.00 c nd nd nd
39 Quercetin-acetyl hexoside 2 nd 0.14 ± 0.02 b 0.08 ± 0.00 c 0.35 ± 0.02 a nd nd nd
40 Kaempferol trihexoside nd 1.27 ± 0.11 a 0.81 ± 0.08 b nd nd nd nd
41 Kaempferol-3-rutinoside 0.92 ± 0.02 a 0.06 ± 0.00 f 0.05 ± 0.01 f 0.54 ± 0.01 b 0.37 ± 0.08 c 0.24 ± 0.03 d 0.15 ± 0.01 e
42 Kaempferol acetyl hexoside 1 nd 1.50 ± 0.15 a 0.32 ± 0.01 b 0.09 ± 0.01 a nd nd nd
43 Kaempferol acetyl hexoside 2 nd 0.07 ± 0.01 a nd 0.33 ± 0.04 a nd nd nd
44 Kaempferol dihexoside nd 0.12 ± 0.01 b 0.44 ± 0.03 a nd nd nd nd
45 Kaempferol pentoside 1 nd nd nd 1.34 ± 0.09 a nd nd nd
46 Kaempferol pentoside 2 nd nd nd 0.12 ± 0.01 a nd nd nd
47 Kaempferol rhamnoside 0.06 ± 0.01 b nd nd nd 0.32 ± 0.08 b 4.43 ± 0.63 a nd
48 Kaempferol hexoside 1 0.01 ± 0.00 e 0.20 ± 0.02 b 0.08 ± 0.00 d 0.27 ± 0.02 a 0.03 ± 0.01 e 0.02 ± 0.00 e 0.14 ± 0.01 c
49 Kaempferol hexoside 2 0.69 ± 0.23 a nd nd 0.38 ± 0.01 b 0.01 ± 0.00 c 0.34 ± 0.04 b nd
50 Kaempferol rhamnosyl hexoside nd nd nd nd 0.02 ± 0.01 b 0.22 ± 0.03 a nd
51 Isorhamnetin hexoside nd 0.01 ± 0.00 c 0.04 ± 0.00 c 0.04 ± 0.00 c 0.28 ± 0.04 b 3.12 ± 0.21 a 0.22 ± 0.00 b
52 Isorhamnetin dihexoside 0.31 ± 0.02 a nd nd nd nd nd nd
53 Isorhamnetin acetyl hexoside 1 nd nd nd nd nd nd 0.98 ± 0.12 a
54 Isorhamnetin acetyl hexoside 2 nd nd nd nd nd nd 0.04 ± 0.00 a
55 Isorhamnetin-3-rutinoside 0.02 ± 0.00 a 0.01 ± 0.00 c 0.02 ± 0.00 b 0.02 ± 0.00 b nd nd nd
56 Myricetin rutinoside nd nd nd nd nd nd 0.004 ± 0.000 a
57 Laricitrin glucuronide 0.07 ± 0.01 a nd nd nd nd nd nd
58 Syringetin hexoside 1 0.02 ± 0.00 b nd nd nd 0.30 ± 0.03 b 5.18 ± 0.62 a nd
59 Syringetin hexoside 2 0.29 ± 0.10 a nd nd nd nd nd nd
60 Syringetin acetyl hexoside 1 nd nd nd nd 1.61 ± 0.41 a 0.78 ± 0.14 b 0.30 ± 0.02 b
61 Syringetin acetyl hexoside 2 nd nd nd nd nd nd 0.07 ± 0.00 a
  Total identified flavonols 5.93 ± 0.05 b 8.71 ± 0.06 b 9.42 ± 0.03 b 8.37 ± 0.02 b 9.29 ± 0.11 b 21.60 ± 0.16 a 2.84 ± 0.02 c
62 Apigenin hexoside nd 0.02 ± 0.00 b 0.01 ± 0.00 b 0.04 ± 0.00 a nd nd nd
  Total identified flavones nd 0.02 ± 0.00 b 0.01 ± 0.00 b 0.04 ± 0.00 a nd nd nd
63 Phloretin xylosylglucoside nd nd nd nd 0.18 ± 0.03 a 0.09 ± 0.02 b nd
64 Phloridzin nd nd nd nd 5.14 ± 0.74 a 5.23 ± 0.52 a nd
65 Trilobatin nd nd nd nd 0.30 ± 0.08 b 1.47 ± 0.20 a nd
  Total identified chalcones nd nd nd nd 5.61 ± 0.28 a 6.80 ± 0.24 a nd
66 Procyanidin dimer 1 nd 0.33 ± 0.03 c 0.52 ± 0.03 c 1.24 ± 0.04 c 1.45 ± 0.42 bc 2.52 ± 0.36 b 7.90 ± 1.48 a
67 Procyanidin dimer 2 nd nd nd 5.07 ± 0.19 a 0.75 ± 0.31 c 2.93 ± 0.47 b 5.30 ± 0.27 a
68 Procyanidin dimer 3 nd nd nd nd 0.02 ± 0.01 b 3.43 ± 0.40 a 0.36 ± 0.02 b
69 Procyanidin dimer 4 nd nd nd nd 0.03 ± 0.01 c 3.43 ± 0.39 b 6.31 ± 0.25 a
70 Procyanidin dimer 5 nd nd nd nd nd nd 1.21 ± 0.08 a
71 Procyanidin dimer 6 nd nd nd nd nd nd 0.50 ± 0.23 a
72 Procyanidin trimer 1 nd 1.18 ± 0.09 d 0.86 ± 0.05 d 11.05 ± 0.30 a 2.15 ± 0.31 c 4.76 ± 0.39 b 0.02 ± 0.00 e
73 Procyanidin trimer 2 nd 2.63 ± 0.18 c 2.68 ± 0.16 c 5.97 ± 0.55 a 0.12 ± 0.05 d 0.07 ± 0.01 d 4.52 ± 0.16 b
74 Procyanidin trimer 3 nd nd nd nd 0.42 ± 0.04 b 8.66 ± 1.08 a 0.02 ± 0.00 b
75 Procyanidin trimer 4 nd nd nd nd 2.33 ± 0.58 a 1.72 ± 0.28 a nd
76 Procyanidin trimer 5 nd nd nd nd nd 4.10 ± 1.60 a nd
77 Procyanidin tetramer nd nd nd nd nd 4.11 ± 1.60 a 2.89 ± 0.18 a
  Total identified condensed tannins nd 4.14 ± 0.10 c 4.06 ± 0.08 c 23.33 ± 0.27 b 7.27 ± 0.22 c 35.73 ± 0.66 a 29.03 ± 0.27 b
  Total identified compounds 16.44 ± 0.06 d 22.05 ± 0.07 d 41.54 ± 0.06 c 65.10 ± 0.09 b 27.62 ± 0.14 d 82.77 ± 0.26 a 46.49 ± 0.09 c
Values represent the mean ± standard deviation of three replicates. Different letters indicate a significant difference among the values in a row (ANOVA, Duncan test, p ≤ 0.05); nd = not detected.

3. LC-DAD-MS Analysis of the Individual Phenolics in the Inflorescences

Detailed studies on the phytochemical profile of Rosaceae plant parts other than the fruits (roots, stems, leaves, and flowers) are very scarce. The identification and quantification of chemical compounds in the inflorescences of Rosaceae contributes to the understanding of the basis of their bioactivity. Just recently, the first results of the phytochemical analyses of the stems, leaves and flowers of Pa were published by Jesus et al. [8]; however, inflorescences were not included. Our study is the first to present a comprehensive LC-DAD-MS polyphenolic profile of selected Rosaceae inflorescences, and it provides a fingerprint for the future quality control of the selected inflorescences.
We identified, in total, 77 phenolic compounds (Table 2, Supplementary Table S1, the supplementary are available online at https://www.mdpi.com/article/10.3390/plants11030271/s1). The representative chromatograms, at different wavelengths, of each of the species are presented in Supplementary Figure S1; the highest number of individual compounds (46), as well as the highest concentration of total identified compounds (82.77 ± 0.26 mg/g DW), were recorded in Mf (Table 2; Supplementary Figure S2). This species also contained the highest concentration of procyanidins, with 35.73 ± 0.66 mg/g DW (Table 1). On the other hand, Pa had the lowest concentration of total identified compounds, 18.05 ± 2.34 mg/g DW. In the inflorescences of this species, we identified 36 compounds, which is significantly more than were identified in its stems, leaves and fruits [8][9][19][22][23], emphasising the wealth of different compounds in inflorescences. However, this was the only sample in which we did not identify individual procyanidins (Table 2). The most individual compounds represented with the highest concentration among the samples were detected in Py (20), representing 49% of all of the identified compounds in this species (Table 2, Supplementary Figure S2).
The individual phenolic with the highest concentration in Pa was di-caffeoylquinic acid (3.12 ± 0.08 mg/g DW), in Ps it was quercetin-acetyl hexoside (4.00 ± 0.40 mg/g DW), in PsKss it was caffeic acid hexoside (15.48 ± 2.23 mg/g DW), in Py and Mf it was procyanidin trimer (11.05 ± 0.30 mg/g DW and 8.66 ± 1.08 mg/g DW, respectively), in Mp it was phloridzin (5.14 ± 0.74 mg/g DW), and in Cj the most represented phenolic was procyanidin dimer (7.90 ± 1.48 mg/g DW) (Table 2). In each of the Prunus inflorescence samples, the concentration of quercetin-3-rutinoside was present with a higher concentration than that of quercetin-3-galactoside, which is opposite to the inflorescence of P. serotina [24], and P. avium inflorescence had a similar concentration of chlorogenic acid to the flowers of P. padus [25].
In Pa, PsKss and Py, the predominant identified compounds were hydroxycinnamic acids in Ps and Mp flavonols, and in Mf and Cj they were procyanidins (Table 1). In Pa, epicatechin was the main flavanol, while catechin was present in a smaller concentration, and the same ratio had already been recorded for fruits as well [23]. Among hydroxybenzoic acids, we identified gallic acid in the concentration of 0.28 ± 0.06 mg/g DW. This was previously found in Pa stems and fruits as well; however, it was not found in leaves and flowers [8].
Mf had a significantly higher concentration of total identified flavonols and procyanidins than other samples, at 21.60 ± 0.16 mg/g DW and 35.73 ± 0.66 mg/g DW, respectively (Table 1). It had far more chlorogenic acid than its whole fruit, flesh or peel [26]. Chalcones were identified in Mf and Mp only, in total concentrations of 6.80 ± 0.24 mg/g DW and 5.61 ± 0.28 mg/g DW, respectively (Table 1). Among them, phloridzin predominated, with 23 ± 0.52 mg/g DW and 5.14 ± 0.74 mg/g DW, respectively. Phloretin xylosylglucoside and trilobatin were represented in inverse proportions in Mf and Mp; in Mf trilobatin predominated with a five-times-higher concentration than in Mp, while in Mp phloretin xylosylglucoside predominated with a two-times-higher concentration than in Mf (Table 1). This suggests that the ratio of phloretin xylosylglucoside and trilobatin could be used as a phytochemical differentiator between these two species.
In Cj, we identified 43 polyphenolic compounds (Table 2), compared to the 20 compounds identified in the fruits [27]. This emphasises the value of inflorescences in the richness and variety of polyphenolic compounds. The representation of flavanols, flavonols and flavanones in Cj inflorescences was similar to that in Chaenomeles maulei fruit juices [28]. The most important polyphenol group was procyanidins (Table 1 and Table 2), which was similar to its leaves [12] and fruits [27].

4. Antidiabetic Activity of the Inflorescences

The antidiabetic activity of the samples was assessed via the inhibition of α-amylase and α-glucosidase, which are enzymes required for carbohydrate digestion. These enzymes are targets not only when attempting to alleviate diabetes but also hyperlipidemia, obesity and caries [29]. The potential of Pa flower, stem and leaf extracts in the inhibition of α-glucosidase has been recognised [8]. Stem extract inhibited α-glucosidase significantly more than extracts of P. avium Saco and Hedelfinger fruits [22], which shows the biopotential of Rosaceae plant parts other than the commonly consumed fruits.
This work is the first report on α-amylase and α-glucosidase inhibition by Rosaceae inflorescences. Each of the tested samples more efficiently inhibited α-glucosidase than α-amylase (Figure 1A,B).
Plants 11 00271 g001 550
Figure 1. Inhibition of (A) α-amylase, (B) α-glucosidase, and (C) 5-lipoxygenase activity by Rosaceae inflorescences’ extracts (0.80 mg/mL, 0.55 mg/mL and 1.45 mg/mL, respectively). The values represent the mean ± standard deviation of three replicates. Different letters indicate a significant difference among the values (ANOVA, Duncan test, p ≤ 0.05). Pa = P. avium, Ps = P. serrulata, PsKss = P. serrulata ‘Kiku Shidare Zakura’, Py = P. yedoensis, Mp = M. purpurea, Mf = M. floribunda, Cj = Chaenomeles japonica, MAL = maltose 0.80 mg/mL, ACAR = acarbose 0.55 mg/mL, NDGA = nordihydroguaiaretic acid 0.15 mg/mL.
The same tendency was recorded with extracts of Chaenomeles fruits as well [20]. However, the fact that the inhibitions of α-amylase, Cj and PsKss were equally efficient to the standard maltose at the same concentration is very interesting and promising. Moreover, both samples even showed a tendency to be more effective than maltose. This emphasizes the high antidiabetic potential of the mentioned inflorescences, and we suggest further in vivo investigations of the antidiabetic activity of these biomaterials and the mechanisms behind this activity. One of the possible intermediates in this α-amylase inhibitory activity could be phenolic compounds. Indeed, phenolic acids and flavonoids bind covalently to α-amylase, forming quinones or lactones that react with nucleophilic groups of the enzyme, thus altering the enzyme’s activity. Cj had the highest concentration of total condensed tannins, 51.68 ± 0.38 mg CE/g DW, while PsKss had the highest concentration of total phenolics, flavonoids and nonflavonoid compounds, at 53.12 ± 0.79 mg GAE/g DW, 38.89 ± 4.04 mg CE/g DW and 31.32 ± 0.71 mg GAE/g DW, respectively (Table 1). Therefore, we assume that, among those measured, these compounds mostly contributed to the inhibition of α-amylase. Moreover, α-amylase showed the highest positive Pearson’s correlation coefficient in its total condensed tannins and total flavonoids, with r = 0.585 and r = 0.458, respectively (Table 3).
Table 3. Pearson’s correlation coefficient (r) between the groups of metabolites, antioxidant capacity, cytotoxicity, and hypoglycemic and anti-inflammatory potential of Rosaceae inflorescences.
  TP TF TNF TT CT SS ABTS FRAP DPPH HepG2 HCT116 HaCaT α-Amyl α-Glucos 5-Lipoxy
TP 1.000                            
TF 0.761 1.000                          
TNF 0.938 0.629 1.000                        
TT 0.471 0.014 0.333 1.000                      
CT −0.065 0.178 −0.367 0.314 1.000                    
SS 0.194 −0.210 0.251 0.262 0.045 1.000                  
ABTS 0.937 0.875 0.908 0.160 −0.140 0.115 1.000                
FRAP 0.873 0.913 0.862 0.016 −0.176 −0.022 0.979 1.000              
DPPH 0.830 0.952 0.762 −0.015 −0.094 −0.170 0.945 0.969 1.000            
HepG2 −0.459 −0.567 −0.396 −0.336 −0.353 0.059 −0.451 −0.489 −0.393 1.000          
HCT116 −0.416 −0.353 −0.327 −0.602 −0.171 0.454 −0.250 −0.274 −0.265 0.689 1.000        
HaCaT −0.148 −0.067 −0.029 −0.670 −0.512 −0.059 −0.004 0.050 0.104 0.729 0.623 1.000      
α-amyl 0.169 0.458 −0.116 −0.038 0.585 −0.064 0.237 0.191 0.377 0.146 0.234 0.117 1.000    
α-glucos 0.077 −0.083 −0.101 0.645 0.627 0.429 −0.065 −0.205 −0.213 −0.312 −0.121 −0.786 0.263 1.000  
5-lipoxy −0.345 0.020 −0.371 −0.530 0.431 0.318 −0.172 −0.114 −0.152 −0.077 0.534 0.154 0.232 0.011 1.000
Values in bold represent strong (0.60–0.79) and very strong (0.80–1.00) correlations.
As in the case of the total condensed tannins, as determined by the spectrophotometric method, which were most represented in the Cj (Table 1), the LC-DAD-MS method also revealed the highest concentrations of individual procyanidins in this species (Table 2). Compared to the other samples, Cj had significantly more procyanidin dimer 1, 2, 4, 5, 6 and procyanidin tetramer, and also 5-p-coumaroylquinic acid 2, naringenin hexoside, isorhamnetin acetyl hexoside 1 and 2, myricetin rutinoside and syringetin acetyl hexoside 2 (Table 2), so we hypothesyze that among these compounds one might potentially find new strong inhibitor/s of α-amylase. Here, we would especially emphasise procyanidins, as they were present in higher amounts—procyanidin dimer 1 even being present at 7.90 ± 1.48 mg/g DW—than the other compounds, and probably mainly contributed to the inhibition of this enzyme. As a support, the significance of tannins for α-amylase inhibition was recognised in work with sorghum as well [30]. An extract of pinhão coat (Araucaria angustifolia) rich in condensed tannins also effectively inhibited α-amylase [29]. On the other hand, in PsKss extract, the predominant compound was caffeic acid hexoside 1 with 15.48 ± 2.23 mg/g DW, which is more than 37% of all of the identified compounds in this species, and this might be one of the key contributors to the inhibition of α-amylase activity. Moreover, recently, fruits from the genus Prunus have been suggested for the preparation of extracts with antidiabetic activities [31].
The potential of Pa fruits to inhibit α-glucosidase is known [22]; however, there are no data on their or other Rosaceae inflorescences’ antidiabetic activity. In our study, Mf and Cj showed a significantly higher rate of α-glucosidase inhibition than the other samples (Figure 1B). Moreover, the inhibition percentages (86.29 ± 3.78% and 83.91 ± 0.48%, respectively) were very close to the value of the standard acarbose at the same concentration, 92.70 ± 1.21% (Figure 1). Compared to the inhibition percentages of common vegetables [32], Rosaceae inflorescences can justifiably be considered relevant natural α-glucosidase inhibitors. Acarbose, a pseudotetrasaccharide, is otherwise a highly effective inhibitor of intestinal α-glucosidases; however, it is not effectively absorbed into the bloodstream, but rather retained in the intestine, and may cause gastrointestinal side effects [29]. Therefore, the plant-based α-glucosidase inhibitors with lower side effects are very welcome. This indicates the high potential of Mf and Cj inflorescences to attenuate hyperglycemia, and should definitely be investigated further in in vivo models. For example, just recently, Kumar et al. [33] revealed that the extract of P. amygdalus seed coat applied to diabetic rats significantly reduced the level of blood glucose, and down-regulated hyperglycemic stress, oxidative stress and hyperlipidaemia. Moreover, they found out that the in vivo antidiabetic activity of the extract was accomplished via the inhibition of dipeptidyl peptidase IV (DPP-IV) protein. This is a hydrolase distributed in various tissues and the circulation, which quickly metabolizes glucagon-like peptide-1 (GLP-1). GLP-1, otherwise, maintains the blood glucose level, supporting insulin secretion and β-cell masses, reducing glucagon secretion, and changing the rate of gastric emptying [34][35]. By inhibiting the DPP-IV, the level of GLP-1 can be maintained; in this way the blood glucose level can be maintained as well. We hypothesize that Mf and Cj inflorescences’ extracts might also affect DPP-IV activity in vivo, and this would be good to test in future. Considering the content of phytochemicals, Mf had the highest concentration of total tannins among the samples, 107.85 ± 1.09 mg CE/g DW, and Cj—as mentioned earlier—had the highest concentration of total condensed tannins and individual identified procyanidins (Table 1, Table 2). Therefore, these groups of compounds were probably responsible for the strong α-glucosidase inhibition. Furthermore, PCA revealed that the total and condensed tannins mostly contributed to the inhibition of α-glucosidase (Figure 2Aii).
Plants 11 00271 g002 550
Figure 2. Principal component analysis of (A) the groups of metabolites, antioxidant capacity, cytotoxicity, antidiabetic and anti-inflammatory potential of Rosaceae inflorescences: (i) score plot separating the inflorescence samples based on the measured groups of metabolites, antioxidant capacity, cytotoxicity, and antidiabetic and anti-inflammatory potential, and (ii) loading plot of the measured variables; (B) the individual identified phenolic compounds in Rosaceae inflorescences: (i) score plot separating the inflorescence samples based on the individual identified phenolic compounds they contain, and (ii) the loading plot of the individual phenolics as variables. Pa = P. avium, Ps = P. serrulata, PsKss = P. serrulata ‘Kiku Shidare Zakura’, Py = P. yedoensis, Mp = M. purpurea, Mf = M. floribunda, Cj = Chaenomeles japonica, TP = total phenolics, TF = total flavonoids, TNF = total nonflavonoids, TT = total tannins, CT = condensed tannins, SS = soluble sugars, ABTS = 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, FRAP = ferric reducing antioxidant power, DPPH = 2,2-diphenyl-1-picrylhydrazyl, 1–77 = numbers related to the individual identified phenolics, as depicted in the Table 2.
The significance of condensed tannins in α-glucosidase inhibition has also been detected in the analysis of P. persica pulp [13]. The individual identified compounds predominating in Cj we have already emphasized, and in Mf the predominant compounds were chlorogenic acid (8.41 ± 0.69 mg/g DW), di-caffeoylquinic acid 2, eriodictyol hexoside 1 and 2, quercetin-3-rhamnoside, quercetin-arabinofuranoside, kaempferol rhamnoside, isorhamnetin hexoside, syringetin hexoside 1, phloridzin, trilobatin, procyanidin dimers, procyanidin trimers, and procyanidin tetramer (Table 2). At the same time, Mf had the highest concentration of total identified compounds, total identified procyanidins, chalcones, flavonols and flavanones (Table 1). As with α-amylase, we would also like to draw attention to the identified procyanidins, especially procyanidin trimer 3, which was present in Mf with more than 10% of the total identified compounds in that species, and probably significantly affected the activity of α-glucosidase. In both of these species, in addition to the identified procyanidins, the predominant compound among the individual compounds was chlorogenic acid, at 8.41 ± 0.69 mg/g DW in Mf and 7.04 ± 0.23 mg/g DW in Cj (Table 2); as such, we assume that these compounds may be responsible for potent α-glucosidase inhibition. Another thing that we detected is that Mf and Cj had high concentrations of procyanidins similar to hawthorn (Crataegus spp.) fruits, which are—due to the preventive activity of these compounds toward oxidative stress after ischemia repercussion injury and myocardial infarction—included in the European pharmacopeia as a complementary treatment for chronic heart failure [36]. Therefore, we think, in the future, that it would be wise to test the potential of Mf and Cj inflorescences’ extracts for the prevention or alleviation of cardiovascular diseases.

5. Anti-Inflammatory Activity of Inflorescences

5-Lipoxygenase catalyses the first step in the biosynthesis of leukotrienes, which are pivotal lipid mediators of inflammation and allergy. The inhibition of this enzyme is one of the strategies to reduce inflammation, and the search for natural inhibitors of 5-lipoxygenase is very active. It is known that a fraction of Py bark suppressed inflammatory chemokines in human HaCaT keratinocytes, and that it might have anti-atopic dermatitis activity [11]. However, the possibility of inflammation suppression with Rosaceae inflorescences has not been tested so far. Therefore, this is the first report on 5-lipoxygenase inhibition by Rosaceae inflorescences.
In our work, Cj was the most efficient species in the inhibition of 5-lipoxygenase, but it was less efficient than the standard nordihydroguaiaretic acid (Figure 1C). Very recently, Turkiewicz et al. [20] tested the inhibition potential of Chaenomeles species and cultivars’ fruits toward 15-lipoxygenase; compared to their results, Cj inflorescences are much more efficient against 5-lipoxygenase, and thus present a promising biomaterial for further analysis. Our analyses showed that only Cj had naringenin-hexoside, isorhamnetin acetyl hexoside 1 and 2, myricetin rutinoside, syringetin acetyl hexoside 2, and procyanidin dimer 5 and 6 (Table 2), and that it had the highest content of total condensed tannins (Table 1), 5-p-coumaroyl-quinic acid 2, and procyanidin dimer 1, 2 and 4 (Table 2). Thus, we assume that these compounds, or at least some of them, were crucial for the anti-inflammatory activity of this sample.

6. Cytotoxic Activity of the Inflorescences

In this study, we evaluated the in vitro anti-proliferative effect of Rosaceae inflorescences on human hepatocellular (HepG2) and colorectal (HCT 116) carcinoma cells, and non-tumorigenic skin keratinocytes (HaCaT). There is evidently a variability in the cell’s metabolic response to the extracts (Table 4).
Table 4. In vitro antiproliferative activity (IC50 expressed in μg/mL) of Rosaceae inflorescence exstracts tested on hepatocellular carcinoma (HepG2), colorectal cancer (HCT 116) and keratinocyte (HaCaT) cell lines.
  Cell Type (IC50 μg/mL)
  HepG2 HCT 116 HaCaT
Prunus avium 300.89 ± 0.21 c A 261.97 ± 13.12 c A 323.84 ± 46.61 c A
Prunus serrulata 473.59 ± 35.69 ab A 517.42 ± 37.10 a A 377.66 ± 34.85 bc B
Prunus serrulata 2018Kiku Shidare Zakura’ 409.71 ± 103.52 b A 464.01 ± 57.31 a A 385.20 ± 7.27 bc A
Prunus yedoensis 508.09 ± 26.28 a A 537.92 ± 43.0 a A 521.64 ± 67.29 a A
Malus purpurea 386.2 ± 19.92 b B 539.66 ± 45.19 a A 461.39 ± 71.56 ab AB
Malus floribunda 445.78 ± 27.42 ab A 361.83 ± 31.19 b B 459.28 ± 43.69 ab A
Chaenomeles japonica 452.48 ± 15.18 ab A 470.66 ± 48.16 a A 473.27 ± 92.54 ab A
Values represent the mean ± standard deviation of three replicates. Different small letters indicate a significant difference among the values in a column, and different capital letters indicate a significant difference among the values in a row (ANOVA, Duncan test, p ≤ 0.05).
The most potent cytotoxic activity shown by Pa toward HCT 116 cell line IC50 was 261.97 ± 13.12 μg/mL, toward HepG2 IC50 was 300.89 ± 0.21 μg/mL, and toward HaCaT cells IC50 was 323.84 ± 46.61 μg/mL (Table 4). Jesus et al. [8] speculated that the high concentration of chlorogenic acid in the leaves of Pa might be the reason of their anticancer activity. However, among the tested samples in our work, Pa was not the one with the highest concentration of this acid (Table 2), and still showed the most potent anticancer activity (Table 4). Therefore, we presume that chlorogenic acid, in itself, may not be crucial for the antiproliferative activity of Pa inflorescences, but in combination with other compounds could act synergistically and enhance the cytotoxicity. Interestingly, Pa stem extracts up to the concentration of 400 μg/mL did not show cytotoxicity toward different cancer cell types, including HepG2, while extracts of the fruits revealed selectivity against colon carcinoma HCT-15 [19]. The cytotoxic effects of isolated catechins are known from both in vitro and in vivo investigations; however, in combinations with the other compounds in the extract, their effects might be different [37]. What surprised us was that Pa had the lowest concentrations of catechins and their oligo- and polymers (Supplementary Figure S3). The cytotoxic activity of these phenolics is well known; however, in the extracts of Pa, Ps, PsKss and Py, some chemical interactions occur between compounds which lead to unexpected effects—the highest cytotoxicity was found in samples with the lowest concentration of catechins, and this was especially emphasized with HaCaT cells, suggesting the cell-based specificity of the extracts as well. Moreover, we detected a strong negative linear correlation between the viability of HaCaT cells, and the content of total tannins (R2 = 0.780) and condensed tannins (R2 = 0.977), and the total identified flavan-3-ols (R2 = 0.952) and procyanidins (R2 = 0.990) among the Prunus samples (Supplementary Figure S3). We hypothesize that these compounds in some way interfere with the cytotoxic effect of Prunus inflorescences’ extracts toward HaCaT. When we looked at the individual components, we detected that epicatechin, procyanidin dimer 2 and procyanidin trimer 1 had strong negative correlations (r = −0.622, r = −0.810, and r = −0.686, respectively) with the HaCaT viability (Supplementary Table S2), so we assume that these compounds might attenuate the cytotoxicity of the Prunus extracts. One possible explanation could be that flavan-3-ols (catechins), and their oligomers and polymers, bind the cytotoxic components of the extracts and act as their antagonists. The higher the concentration of the total and condensed tannins, and the total identified procyanidins and flavanols in Prunus inflorescences, the higher the number of viable HaCaT cells. This is useful information for the possible application of the tested Prunus extracts in dermal wound healing.
Ps and PsKss showed similar cytotoxic potentials to Pa toward HaCaT cells. Cell-specific cytotoxicity levels were ascertained for Ps (HaCaT was most susceptible), Mp (HepG2 was most sensitive), and Mf (HCT 116 was most sensitive) (Table 4).
So far, the anticancer properties of the fruits and stems of Pa have been investigated against five human cancer cell lines, and only the extract of the fruits showed activity against colon carcinoma HCT-15 [19]. Because fruits contain anthocyanins, and stems do not, the presumption is that they might be the key to the anticancer properties of Pa fruits. The inflorescences have not been tested so far.
A very strong positive correlation was found between cytotoxic activity against HepG2 and di-caffeoylquinic acid 3, isorhamnetin dihexoside, laricitrin glucuronide and syringetin hexoside 2 (Supplementary Table S1), all of which are predominantly present in Pa. As such, these compounds might be responsible for the inhibition of cell proliferation, and their cytotoxic potential should be investigated further. The cytotoxic activity toward HCT116 was very strongly correlated with di-caffeoylquinic acid 3,5-feruloylquinic acid, isorhamnetin dihexoside, laricitrin glucuronide and syringetin hexoside 2, which were again predominant in Pa. On the other hand, caffeic acid dihexoside was the only compound that was very strongly correlated with an antiproliferative effect on HaCaT cells (Supplementary Table S2).

7. Statistical Analysis

The principal component analysis (PCA) based on the measured groups of metabolites and the antioxidant capacity (Table 1), cytotoxicity (Table 4), and hypoglycemic and anti-inflammatory potential (Figure 1) explained 64.68% of the total variation among the samples, where PC1 accounted for 41.71% of the variance and PC2 accounted for 22.97% (Figure 2Ai). The samples were separated into three groups: Ps and PsKss formed one group; Mp, Cj, Py and Mf formed the other; and Pa was separated alone as the most specific sample (Figure 2Ai). The variables that mostly contributed to the group of Ps and PsKss were antioxidant capacity (ABTS, FRAP and DPPH), total phenolics, flavonoids, and nonflavonoid compounds (Figure 2Aii). The separation of Pa was due to the cytotoxic activity toward all of the three tested cell types. The group of Mp, Cj, Py and Mf mostly contributed tannins, soluble sugars, and the inhibition of α-glucosidase.
Based on the individual identified phenolics, the samples were similarly grouped: the only difference was in the grouping of Py, which was closer to Pa, and they grouped together (Figure 2Bi). This suggests that the representation of the individual identified phenolics and the bioactivity of the tested Rosaceae inflorescences are analogously distributed between all of the samples, except for Py. We also noticed that Cj, based on the measured parameters, was closer to the Malus than to the Prunus samples.
HC analysis, an algorithm that creates a dendrogram showing the hierarchical relationships between different datasets, showed the degree of similarities/dissimilarities between the samples. Based on their groups of metabolites, antioxidant capacity, cytotoxicity, and hypoglycemic and anti-inflammatory potential, Ps and PsKss were the most similar samples to each other. Other samples were close to them, while Pa was the most distant from all of the samples (Supplementary Figure S4A). These results indicate that greater genetic similarity does not imply a greater similarity in biological effects (Supplementary Figure S4A).
Based on their individual identified phenolic compounds, the samples were, as expected, more similar (closer) to each other (Supplementary Figure S4B); the most similar were Pa and Ps, while the most distant from them was Py.
Pearson’s correlation coefficient (r) between the groups of metabolites, antioxidant capacity, cytotoxicity, and hypoglycemic and anti-inflammatory potential of the samples revealed a strong positive correlation between antioxidant capacity (ABTS, FRAP and DPPH) and the total phenolics, flavonoids and nonflavonoids (Table 3). The inhibition of α-glucosidase was strongly positively correlated with cytotoxicity toward HaCaT cells (Table 3). Cytotoxicity toward HepG2 was positively correlated with cytotoxicity toward both HCT 116 and HaCaT cells. Cytotoxic activities toward HCT 116 and HaCaT were also strongly correlated. The total tannins exhibited a strong negative correlation with cytotoxicity toward HCT 116 and HaCaT cells. This suggests that a removal of tannins from the extracts might increase the cytotoxic effect/s of the samples toward HCT 116 and HaCaT, along with their inhibition of α-glucosidase, which is worthy of further investigations. The inhibition of α-glucosidase was strongly positively correlated with the total and condensed tannins, which indicates that these compounds might be responsible for the inhibition of this enzyme. These results are in accordance with previous observations regarding the tannin effect on glucosidase activity [38].
As far as individual compounds are concerned, according to Pearson’s correlation coefficient, the only compound that was very strongly correlated with the inhibition of α-glucosidase was chlorogenic acid (Supplementary Table S2); as such, we suggest the further investigation of the antidiabetic potential of this phenolic compound. The results from all three antioxidant methods strongly correlated with caffeic acid hexoside 1, 3-caffeoylquinic acid, 4-caffeoylquinic acid, 3-p-coumaroylquinic acid, p-coumaric acid hexoside 2, catechin, quercetin-hexoside pentoside, quercetin-3-glucoside, kaempferol dihexoside and kaempferol trihexoside. Therefore, we assume that these compounds mostly contributed to the antioxidant activity of the inflorescences. The cytotoxic activity toward HepG2, HCT 116 and HaCaT strongly correlated with di-caffeoylquinic acid 3,3-feruloylquinic acid, 5-feruloylquinic acid, isorhamnetin dihexoside, laricitrin glucuronide and syringetin hexoside 2. The individual compounds that strongly correlated with the inhibition of both antidiabetic enzymes, α-amylase and α-glucosidase, were procyanidin dimer 1 and 4. With the inhibition of α-amylase, the only strong correlations were naringenin hexoside, isorhamnetin acetyl hexoside 1 and 2, myricetin rutinoside, syringetin acetyl hexoside 2, and procyanidin dimer 1, 4, 5 and 6. The inhibition of α-glucosidase strongly correlated with chlorogenic acid; procyanidin dimer 1, 2, 4; and procyanidin tetramer.
Based on the results, we propose the consideration of Prunus, Malus and Chaenomeles inflorescences as low-sugar plant foods rich in polyphenolic bioactive compounds, or at least as functional additives to regular food that could improve human health. In particular, we propose Cj inflorescences for further in vivo studies of their antidiabetic and anti-inflammatory activity, and for possible use as a functional food. Prunus inflorescences would be excellent candidates for further analyses of the influence of monomers, oligomers and polymers of flavanols on HaCaT cells’ proliferation. Finally, PsKss is a material with a respectable amount of total phenolics and flavonoids that shows strong antioxidant activity and α-amylase inhibition; therefore, it is worthy of further in vitro and in vivo investigations.

References

  1. Orlando, G.; Recinella, L.; Chiavaroli, A.; Brunetti, L.; Leone, S.; Carradori, S.; Di Simone, S.; Ciferri, M.C.; Zengin, G.; Ak, G.; et al. Water extract from inflorescences of industrial hemp futura 75 variety as a source of anti-inflammatory, anti-proliferative and antimycotic agents: Results from in silico, in vitro and ex vivo studies. Antioxidants 2020, 9, 437.
  2. Lau, B.F.; Kong, K.W.; Leong, K.H.; Sun, J.; He, X.; Wang, Z.; Mustafa, M.R.; Ling, T.C.; Ismail, A. Banana inflorescence: Its bio-prospects as an ingredient for functional foods. Trends Food Sci. Technol. 2020, 97, 14–28.
  3. Sarv, V.; Venskutonis, P.R.; Bhat, R. The Sorbus spp.—Underutilised plants for foods and nutraceuticals: Review on polyphenolic phytochemicals and antioxidant potential. Antioxidants 2020, 9, 813.
  4. El Euch, S.K.; Bouajila, J.; Bouzouita, N. Chemical composition, biological and cytotoxic activities of Cistus salviifolius flower buds and leaves extracts. Ind. Crops Prod. 2015, 76, 1100–1105.
  5. Lee, E.J.; Kim, J.S.; Kim, H.P.; Lee, L.-H.; Kang, S.S. Phenolic constituents from the flower buds of Lonicera japonica and their 5-lipoxygenase inhibitory activities. Food Chem. 2010, 120, 134–139.
  6. Ferreres, F.; Andrade, C.; Gomes, N.G.; Andrade, P.B.; Gil-Izquierdo, A.; Pereira, D.M.; Suksungworn, R.; Duangsrisai, S.; Videira, R.A.; Valentão, P. Valorisation of kitul, an overlooked food plant: Phenolic profiling of fruits and inflorescences and assessment of their effects on diabetes-related targets. Food Chem. 2021, 342, 128323.
  7. Olszewska, M.A.; Michel, P. Antioxidant activity of inflorescences, leaves and fruits of three Sorbus species in relation to their polyphenolic composition. Nat. Prod. Res. 2009, 23, 1507–1521.
  8. Jesus, F.; Gonçalves, A.C.; Alves, G.; Silva, L.R. Health benefits of Prunus avium plant parts: An unexplored source rich in phenolic compounds. Food Rev. Int. 2020.
  9. Acero, N.; Gradillas, A.; Beltran, M.; García, A.; Muñoz Mingarro, D. Comparison of phenolic compounds profile and antioxidant properties of different sweet cherry (Prunus avium L.) varieties. Food Chem. 2019, 279, 260–271.
  10. Usenik, V.; Fabčič, J.; Štampar, F. Sugars, organic acids, phenolic composition and antioxidant activity of sweet cherry (Prunus avium L.). Food Chem. 2008, 107, 185–192.
  11. Kang, G.-J.; Lee, H.-J.; Yoon, W.-J.; Yang, E.-J.; Park, S.-S.; Kang, H.-K.; Park, M.-H.; Yoo, E.-S. Prunus yedoensis inhibits the inflammatory chemokines, MDC and TARC, by regulating the STAT1-signaling pathway in IFN-γ-stimulated HaCaT human keratinocytes. Biomol. Ther. 2008, 16, 394–402.
  12. Teleszko, M.; Wojdyło, A. Comparison of phenolic compounds and antioxidant potential between selected edible fruits and their leaves. J. Funct. Foods 2015, 14, 736–746.
  13. Zhang, X.; Su, M.; Du, J.; Zhou, H.; Li, X.; Li, X.; Ye, Z. Comparison of phytochemical differences of the pulp of different peach (Prunus persica (L.) Batsch) cultivars with alpha-glucosidase inhibitory activity variations in China using UPLC-Q-TOF/MS. Molecules 2019, 24, 1968.
  14. Wojdyło, A.; Nowicka, P.; Grimalt, M.; Legua, P.; Almansa, M.S.; Amorós, A.; Carbonell-Barrachina, A.; Hernández, F. Polyphenol compounds and biological activity of caper (Capparis spinosa L.) flowers buds. Plants 2019, 8, 539.
  15. Borneo, R.; León, A.E.; Aguirre, A.; Ribotta, P.; Cantero, J.J. Antioxidant capacity of medicinal plants from the Province of Córdoba (Argentina) and their in vitro testing in a model food system. Food Chem. 2009, 112, 664–670.
  16. Afonso, S.; Oliveira, I.V.; Meyer, A.S.; Aires, A.; Saavedra, M.J.; Gonçalves, B. Phenolic profile and bioactive potential of stems and seed kernels of sweet cherry fruit. Antioxidants 2020, 9, 1295.
  17. Yang, S.; Meng, Z.; Li, Y.; Chen, R.; Yang, Y.; Zhao, Z. Evaluation of physiological characteristics, soluble sugars, organic acids and volatile compounds in ‘Orin’ apples (Malus domestica) at different ripening stages. Molecules 2021, 26, 807.
  18. Walker, R.P.; Battistelli, A.; Bonghi, C.; Drincovich, M.F.; Falchi, R.; Lara, M.V.; Moscatello, S.; Vizzotto, G.; Famiani, F. Non-structural carbohydrate metabolism in the flesh of stone fruits of the genus Prunus (Rosaceae)—A review. Front. Plant Sci. 2020, 11, 549921.
  19. Bastos, C.; Barros, L.; Dueñas, M.; Calhelha, R.C.; Queiroz, M.J.R.; Santos-Buelga, C.; Ferreira, I.C. Chemical characterisation and bioactive properties of Prunus avium L.: The widely studied fruits and the unexplored stems. Food Chem. 2015, 173, 1045–1053.
  20. Turkiewicz, I.P.; Wojdyło, A.; Tkacz, K.; Nowicka, P.; Golis, T.; Bąbelewski, P. ABTS On-line antioxidant, α-amylase, α-glucosidase, pancreatic lipase, acetyl-and butyrylcholinesterase inhibition activity of Chaenomeles fruits determined by polyphenols and other chemical compounds. Antioxidants 2020, 9, 60.
  21. Fang, L.; Meng, W.; Min, W. Phenolic compounds and antioxidant activities of flowers, leaves and fruits of five crabapple cultivars (Malus Mill. species). Sci. Hortic. 2018, 235, 460–467.
  22. Gonçalves, A.C.; Bento, C.; Silva, B.M.; Silva, L.R. Sweet cherries from Fundão possess antidiabetic potential and protect human erythrocytes against oxidative damage. Food Res. Int. 2017, 95, 91–100.
  23. Gonçalves, B.; Landbo, A.K.; Knudsen, D.; Silva, A.P.; Moutinho-Pereira, J.; Rosa, E.; Meyer, A.S. Effect of ripeness and postharvest storage on the phenolic profiles of cherries (Prunus avium L.). J. Agric. Food Chem. 2004, 52, 523–530.
  24. Olszewska, M. Quantitative HPLC analysis of flavonoids and chlorogenic acid in the leaves and inflorescences of Prunus serotina Ehrh. Acta Chromatogr. 2007, 19, 253–269.
  25. Olszewska, M.A.; Kwapisz, A. Metabolite profiling and antioxidant activity of Prunus padus L. flowers and leaves. Nat. Prod. Res. 2011, 25, 1115–1131.
  26. Coklar, H.; Akbulut, M.; Alhassan, I.; Kirpitci, Ş.; Korkmaz, E. Organic acids, sugars, phenolic compounds and antioxidant activity of Malus floribunda coccinella fruit, peel and flesh. Acta Sci. Pol. Hortorum Cultus 2018, 17, 47–59.
  27. Du, H.; Wu, J.; Li, H.; Zhong, P.-X.; Xu, Y.-J.; Li, C.-H.; Ji, K.-X.; Wang, L.-S. Polyphenols and triterpenes from Chaenomeles fruits: Chemical analysis and antioxidant activities assessment. Food Chem. 2013, 141, 4260–4268.
  28. Valcheva-Kuzmanova, S.V.; Denev, P.N.; Ognyanov, M.H. Chemical composition and antioxidant activity of Chaenomeles maulei fruit juice. J. Biomed. Clin. Res. 2018, 11, 41–48.
  29. Da Silva, S.M.; Koehnlein, E.A.; Bracht, A.; Castoldi, R.; de Morais, G.R.; Baesso, M.L.; Peralta, R.; de Souza, C.G.M.; de Sá-Nakanishi, A.B.; Peralta, R.M. Inhibition of salivary and pancreatic α-amylases by a pinhão coat (Araucaria angustifolia) extract rich in condensed tannin. Food Res. Int. 2014, 56, 1–8.
  30. Mkandawire, N.L.; Kaufman, R.C.; Bean, S.R.; Weller, C.L.; Jackson, D.S.; Rose, D.J. Effects of sorghum (Sorghum bicolor (L.) Moench) tannins on alpha-amylase activity and in vitro digestibility of starch in raw and processed flours. J. Agric. Food Chem. 2013, 61, 4448–4454.
  31. Papoutsis, K.; Zhang, J.; Bowyer, M.C.; Brunton, N.; Gibney, E.R.; Lyng, J. Fruit, vegetables, and mushrooms for the preparation of extracts with α-amylase and α-glucosidase inhibition properties: A review. Food Chem. 2021, 338, 128119.
  32. Assefa, S.T.; Yang, E.Y.; Chae, S.Y.; Song, M.; Lee, J.; Cho, M.C.; Jang, S. Alpha glucosidase inhibitory activities of plants with focus on common vegetables. Plants 2020, 9, 2.
  33. Kumar, V.; Sachan, R.; Rahman, M.; Sharma, K.; Al-Abassi, F.A.; Anwar, F. Prunus amygdalus extract exert antidiabetic effect via inhibition of DPP-IV: In-silico and in-vivo approaches. J. Biomol. Struct. Dyn. 2021, 39, 4160–4174.
  34. Meier, J.J. GLP-1 receptor agonists for individualized treatment of type 2 diabetes mellitus. Nat. Rev. Endocrinol. 2012, 8, 728–742.
  35. Seino, Y.; Fukushima, M.; Yabe, D. GIP and GLP-1, the two incretin hormones: Similarities and differences. J. Diabetes Investig. 2010, 1, 8–23.
  36. Wang, T.K.; Xu, S.; Li, S.; Zhang, Y. Proanthocyanidins should be a candidate in the treatment of cancer, cardiovascular diseases and lipid metabolic disorder. Molecules 2020, 25, 5971.
  37. Xu, Y.Q.; Gao, Y.; Granato, D. Effects of epigallocatechin gallate, epigallocatechin and epicatechin gallate on the chemical and cell-based antioxidant activity, sensory properties, and cytotoxicity of a catechin-free model beverage. Food Chem. 2021, 339, 128060.
  38. Muccilli, V.; Cardullo, N.; Spatafora, C.; Cunsolo, V.; Tringali, C. α-Glucosidase inhibition and antioxidant activity of an oenological commercial tannin. Extraction, fractionation and analysis by HPLC/ESI-MS/MS and 1H NMR. Food Chem. 2017, 215, 50–60.
More
© Text is available under the terms and conditions of the Creative Commons Attribution (CC BY) license; additional terms may apply. By using this site, you agree to the Terms and Conditions and Privacy Policy.
Upload a video for this entry
Information
Subjects: Agriculture, Dairy & Animal Science
Contributor MDPI registered users' name will be linked to their SciProfiles pages. To register with us, please refer to https://encyclopedia.pub/register :
Ivana Šola
View Times: 394
Revisions: 2 times (View History)
Update Date: 21 Jan 2022
Table of Contents
    1000/1000
    Hot Most Recent
    ScholarVision Creations
    Feedback