1. Please check and comment entries here.
Table of Contents

    Topic review

    Thymoquinone and Curcumin

    View times: 12
    Submitted by: Ali El-Far


    D-galactose (D-gal) administration causes oxidative disorder and is widely utilized in aging animal models. Therefore, subcutaneously injected D-gal at 200 mg/kg BW dose to assess the potential preventive effect of thymoquinone (TQ) and curcumin (Cur) against the oxidative alterations induced by D-gal. Other than the control, vehicle, and D-gal groups, the TQ and Cur treated groups were orally supplemented at 20 mg/kg BW of each alone or combined. TQ and Cur effectively suppressed the oxidative alterations induced by D-gal in brain and heart tissues. The TQ and Cur combination significantly decreased the elevated necrosis in the brain and heart by D-gal. It significantly reduced brain caspase 3, calbindin, and calcium-binding adapter molecule 1 (IBA1), heart caspase 3, and BCL2. Expression of mRNA of the brain and heart TP53, p21, Bax, and CASP-3 were significantly downregulated in the TQ and Cur combination group along with upregulation of BCL2 in comparison with the D-gal group. Data suggested that the TQ and Cur combination is a promising approach in aging prevention. 

    1. Introduction

    Aging is a deteriorative process that occurs mainly due to oxidative stress, leading to numerous oxidative stress-associated diseases because of the accumulation of reactive oxygen species (ROS) and reduced antioxidant capability [1][2]. A D-galactose (D-gal)-induced aging model is a commonly utilized model to investigate anti-aging drugs [3]. When D-gal accumulates in the body, it can react with the free amines of amino acids in proteins and peptides, forming advanced glycation end products (AGEs) [4]. Consequently, AGEs interact with specific receptors (RAGE) in many cell types and induce the activation of the downstream nuclear factor kappa-B (NF-κB), and other signaling pathways, resulting in ROS generation, which could accelerate the aging process [4][5]. The elevated ROS and reactive nitrogen species (RNS), including superoxide anion and nitric oxide, lead to cellular damages in protein, lipid, and DNA that are able to favor the development of different diseases, including tumors, neurodegenerative disorders, aging, and an inflammatory processes [6][7][8][9].
    Natural compounds act as preventive antioxidant agents against different age-associated alterations [10]. Thymoquinone (TQ) is an active compound of Nigella sativa seeds with diverse biological activities such as antioxidant, antitoxic, anti-inflammatory, antidiabetic, and anticancer activities (Figure 1) [11][12][13][14][15]. TQ showed a positive elevation in liver glutathione levels and enhancement of total oxidant status of blood in a rat model with carbon tetrachloride-induced hepatotoxicity [16]. Also, TQ protects cardiac muscles against diabetic oxidative stress by upregulation of nuclear factor-erythroid-2-related factor 2 (Nrf2), which improved the antioxidant potential of the cardiac muscles and alleviated the inflammatory process [12]. Moreover, TQ alleviates the testicular damage in diabetic rats through its powerful antioxidant and hypoglycemic effects [11]. Additionally, TQ shows a regenerative potential for treating damaged peripheral nerves [17].
    Curcumin (Cur) is a yellow pigment obtained from Curcuma longa, commonly used as a spice and food-coloring agent (Figure 1). It has preventive or putative therapeutic properties because of its anti-inflammatory, antioxidant, anti-aging, and anticancer potential [18][19][20][21][22][23][24].

    2. Current Insights

    Aging (senescence) is the loss of organ and tissue function gradually with time [25]. The losses of age-associated functions are because of the cumulation of oxidative damage macromolecules (proteins, DNA, and lipids) by ROS and RNS [26]. Senescent cells pile through aging and have been involved in enhancing various age-related diseases [27]. Senescence inducers led to upregulation of p53, which elevated the cyclin-dependent kinase inhibitor p21(WAF1/CIP1), mainly mediating G1 growth arrest [28].
    The mechanism D-gal inducing aging is well recognized and based on generation of ROS and RNS that induce inflammation and apoptosis of different body cells [4][5]. In the current study, we assessed the aging and apoptotic markers due to D-gal administration and the protective role of TQ, Cur and their combination. D-gal significantly upregulated p21 and TP53, leading to aging oxidative alterations in brain and heart tissues. D-gal-induced upregulation of p21 and p53 in mouse [29] and rat [30] models treated by D-gal. Moreover, western blot results revealed upregulation of the p53/p21 signaling pathway in mice’s hippocampus [31].
    TQ, Cur, and their combination significantly downregulated the increased expression of p21 and TP53 because of D-gal. Also, TQ is responsible for apoptosis induction in colorectal cancer by inhibiting the p53-dependent CHEK1 [32]. Some rationales suggest that curcumin’s anti-aging function is due to its ability to postpone cellular senescence in cells building the vasculature [32].
    An elevated ROS level accompanies humans’ aging and higher animals in mitochondria, inducing apoptosis, lowering the functioning cells’ number [33]. D-gal-stimulated brain aging exhibited changes in cognitive function and brain mitochondria [34]. Also, hypertrophy of the myocytes and myocytes’ loss are characteristic foraging in the mammalian heart [35]. During heart failure and normal heart aging, necrosis and apoptosis mechanisms are involved in myocyte cell loss [36][37]. In the present study, D-gal induced necrosis and apoptosis of brain and heart tissues monitored by upregulation of apoptotic (CASP-3 and Bax genes and caspase 3 protein) and downregulation of antiapoptotic (Blc2 gene and protein) markers. In the same context, D-gal induced significant decreases in the Bax/Bcl-2 ratio and caspase-3 in mice’s brains [38] and rats [39]. Also, D-gal markedly lowered the Bax/Bcl-2 ratio and caspase-3 in aging rats’ cardiomyocytes [40]. In contrast, TQ and Cur attenuated the necrotic and apoptotic alterations of rats’ brains and hearts, especially their combination. Similarly, Abulfadl et al. [41] stated that TQ prevented D-gal/AlCl3-induced cognitive decline by promoting synaptic plasticity and cholinergic function and suppressing neuronal apoptosis oxidative damage and neuroinflammation in rats. Also, curcumin minimized the alterations induced in Purkinje cells [42] and cleaved caspase-3 expression [43] in rats due to D-gal.
    Calcium has a pivotal role in the neurodegeneration process and has an essential role as an intracellular signaling mediator [44]. Therefore, multiple injury pathways meet to stimulate an extra increase in intracellular calcium levels, inducing a series of caspases leading to the apoptosis onset. So, the calcium homeostasis maintenance within neurons is essential to their health, including many mechanisms [45]. Calbindin is a calcium-binding protein that protects neurons against damage caused by excessive Ca2+ elevation [46]. Thus, in the current investigation, D-gal induced reduced brain calbindin in rats leading to activation of caspase 3 that induced apoptosis of brain tissue, while TQ, Cur, and their combinations attenuated the calbindin reduction due to D-gal. There is no published article regarding the influence of D-gal or TQ on brain calbindin expression. At the same time, curcuminoid submicron particle consumption inverted spatial memory deficits and the loss of calbindin in the hippocampus of the Alzheimer’s disease mouse model [47].
    IBA1 is a cytoskeleton protein localized only in macrophages and microglia [48]. The IBA1 expression is upregulated in stimulated microglia after ischemia [49], peripheral nerve injury [50][51], and many brain diseases [52]. In the present investigation, we recognized a marked elevation in IBA1 expression in brain tissue. Similarly, D-gal significantly increased the neuroinflammatory marker, IBA1, in the mouse brain [53][54]. Conversely, TQ and Cur and their combinations significantly reduced the elevated IBA1 in response to D-gal.

    The entry is from 10.3390/ijms22136839


    1. Chen, W.K.; Tsai, Y.L.; Shibu, M.A.; Shen, C.Y.; Chang-Lee, S.N.; Chen, R.J.; Yao, C.H.; Ban, B.; Kuo, W.W.; Huang, C.Y. Exercise training augments Sirt1-signaling and attenuates cardiac inflammation in D-galactose induced-aging rats. Aging 2018, 10, 4166–4174.
    2. Davalli, P.; Mitic, T.; Caporali, A.; Lauriola, A.; D’Arca, D. ROS, Cell Senescence, and Novel Molecular Mechanisms in Aging and Age-Related Diseases. Oxid. Med. Cell. Longev. 2016, 2016, 1–18.
    3. Zhao, F.; Zhou, Y.; Gao, L.; Qin, X.; Du, G. [Advances in the study of the rat model of aging induced by D-galactose]. Yao Xue Xue Bao 2017, 52, 347–354.
    4. Hegab, Z. Role of advanced glycation end products in cardiovascular disease. World J. Cardiol. 2012, 4, 90.
    5. Frimat, M.; Daroux, M.; Litke, R.; Nevière, R.; Tessier, F.J.; Boulanger, E. Kidney, heart and brain: Three organs targeted by ageing and glycation. Clin. Sci. 2017, 131, 1069–1092.
    6. Vivarelli, S.; Falzone, L.; Basile, M.S.; Candido, S.; Libra, M. Nitric Oxide in Hematological Cancers: Partner or Rival? Antioxid. Redox Signal. 2021, 34, 383–401.
    7. Singh, A.; Kukreti, R.; Saso, L.; Kukreti, S. Oxidative Stress: A Key Modulator in Neurodegenerative Diseases. Molecules 2019, 24, 1583.
    8. Chen, X.; Guo, C.; Kong, J. Oxidative stress in neurodegenerative diseases. Neural Regen. Res. 2012, 7, 376–385.
    9. Sahni, S.; Hickok, J.R.; Thomas, D.D. Nitric oxide reduces oxidative stress in cancer cells by forming dinitrosyliron complexes. Nitric Oxide Biol. Chem. 2018, 76, 37–44.
    10. Ma, Y.; Ma, B.; Shang, Y.; Yin, Q.; Wang, D.; Xu, S.; Hong, Y.; Hou, X.; Liu, X. Flavonoid-Rich Ethanol Extract from the Leaves of Diospyros kaki Attenuates D-Galactose-Induced Oxidative Stress and Neuroinflammation-Mediated Brain Aging in Mice. Oxid. Med. Cell. Longev. 2018, 2018, 1–12.
    11. Atta, M.S.; Almadaly, E.A.; El-Far, A.H.; Saleh, R.M.; Assar, D.H.; Al Jaouni, S.K.; Mousa, S.A. Thymoquinone Defeats Diabetes-Induced Testicular Damage in Rats Targeting Antioxidant, Inflammatory and Aromatase Expression. Int. J. Mol. Sci. 2017, 18, 919.
    12. Atta, M.S.; El-Far, A.H.; Farrag, F.A.; Abdel-Daim, M.M.; Al Jaouni, S.K.; Mousa, S.A. Thymoquinone attenuates cardiomyopathy in streptozotocin-treated diabetic rats. Oxid. Med. Cell. Longev. 2018, 2018, 1–10.
    13. El-Far, A. Thymoquinone Anticancer Discovery: Possible Mechanisms. Curr. Drug Discov. Technol. 2015, 12, 80–89.
    14. El-Far, A.H.; Al Jaouni, S.K.; Li, W.; Mousa, S.A. Protective roles of thymoquinone nanoformulations: Potential nanonutraceuticals in human diseases. Nutrients 2018, 10, 1369.
    15. El-Far, A.H.; Korshom, M.A.; Mandour, A.A.; El-Bessoumy, A.A.; El-Sayed, Y.S. Hepatoprotective efficacy of Nigella sativa seeds dietary supplementation against lead acetate-induced oxidative damage in rabbit—Purification and characterization of glutathione peroxidase. Biomed. Pharmacother. 2017, 89, 711–718.
    16. Demiroren, K.; Basunlu, M.T.; Erten, R.; Cokluk, E. A comparison of the effects of thymoquinone, silymarin and N-acetylcysteine in an experimental hepatotoxicity. Biomed. Pharmacother. 2018, 106, 1705–1712.
    17. Üstün, R.; Oğuz, E.K.; Şeker, A.; Korkaya, H. Thymoquinone protects DRG neurons from axotomy-induced cell death. Neurol. Res. 2018, 40, 930–937.
    18. Menon, V.P.; Sudheer, A.R. Antioxidant and anti-inflammatory properties of curcumin. Adv. Exp. Med. Biol. 2007, 595, 105–125.
    19. Samarghandian, S.; Azimi-Nezhad, M.; Farkhondeh, T.; Samini, F. Anti-oxidative effects of curcumin on immobilization-induced oxidative stress in rat brain, liver and kidney. Biomed. Pharmacother. 2017, 87, 223–229.
    20. Chakraborty, M.; Bhattacharjee, A.; Kamath, J.V. Cardioprotective effect of curcumin and piperine combination against cyclophosphamide-induced cardiotoxicity. Indian J. Pharmacol. 2017, 49, 65–70.
    21. Banji, D.; Banji, O.J.F.F.; Dasaroju, S.; Annamalai, A.R.R. Piperine and curcumin exhibit synergism in attenuating d-galactose induced senescence in rats. Eur. J. Pharmacol. 2013, 703, 91–99.
    22. Kunnumakkara, A.B.; Diagaradjane, P.; Anand, P.; Kuzhuvelil, H.B.; Deorukhkar, A.; Gelovani, J.; Guha, S.; Krishnan, S.; Aggarwal, B.B. Curcumin sensitizes human colorectal cancer to capecitabine by modulation of cyclin D1, COX-2, MMP-9, VEGF and CXCR4 expression in an orthotopic mouse model. Int. J. Cancer 2009, 125, 2187–2197.
    23. Hu, S.; Xu, Y.; Meng, L.; Huang, L.; Sun, H. Curcumin inhibits proliferation and promotes apoptosis of breast cancer cells. Exp. Ther. Med. 2018, 16, 1266–1272.
    24. Shakibaei, M.; Mobasheri, A.; Lueders, C.; Busch, F.; Shayan, P.; Goel, A. Curcumin enhances the effect of chemotherapy against colorectal cancer cells by inhibition of NF-κB and Src protein kinase signaling pathways. PLoS ONE 2013, 8, e57218.
    25. Flatt, T. A new definition of aging? Front. Genet. 2012, 3, 148.
    26. Beckman, K.B.; Ames, B.N. The free radical theory of aging matures. Physiol. Rev. 1998, 78, 547–581.
    27. Regulski, M.J. Cellular Senescence: What, Why, and How. Wounds 2017, 29, 168–174.
    28. Galanos, P.; Vougas, K.; Walter, D.; Polyzos, A.; Maya-Mendoza, A.; Haagensen, E.J.; Kokkalis, A.; Roumelioti, F.M.; Gagos, S.; Tzetis, M.; et al. Chronic p53-independent p21 expression causes genomic instability by deregulating replication licensing. Nat. Cell Biol. 2016, 18, 777–789.
    29. Wang, Z.L.; Chen, L.B.; Qiu, Z.; Chen, X.B.; Liu, Y.; Li, J.; Wang, L.; Wang, Y.P. Ginsenoside Rg1 ameliorates testicular senescence changes in D-gal-induced aging mice via anti-inflammatory and antioxidative mechanisms. Mol. Med. Rep. 2018, 17, 6269–6276.
    30. Sun, J.; Jing, S.; Jiang, R.; Wang, C.; Zhang, C.; Chen, J.; Li, H. Metabolomics study of the therapeutic mechanism of schisandra chinensis lignans on aging rats induced by D-galactose. Clin. Interv. Aging 2018, 13, 829–841.
    31. Li, Q.; Zeng, J.; Su, M.; He, Y.; Zhu, B. Acetylshikonin from Zicao attenuates cognitive impairment and hippocampus senescence in D-galactose-induced aging mouse model via upregulating the expression of SIRT1. Brain Res. Bull. 2018, 137, 311–318.
    32. Gali-Muhtasib, H.; Kuester, D.; Mawrin, C.; Bajbouj, K.; Diestel, A.; Ocker, M.; Habold, C.; Foltzer-Jourdainne, C.; Schoenfeld, P.; Peters, B.; et al. Thymoquinone triggers inactivation of the stress response pathway sensor CHEK1 and contributes to apoptosis in colorectal cancer cells. Cancer Res. 2008, 68, 5609–5618.
    33. Isaev, N.K.; Genrikhs, E.E.; Oborina, M.V.; Stelmashook, E.V. Accelerated aging and aging process in the brain. Rev. Neurosci. 2018, 29, 233–240.
    34. Shwe, T.; Pratchayasakul, W.; Chattipakorn, N.; Chattipakorn, S.C. Role of D-galactose-induced brain aging and its potential used for therapeutic interventions. Exp. Gerontol. 2018, 101, 13–36.
    35. Olivetti, G.; Melissari, M.; Capasso, J.M.; Anversa, P. Cardiomyopathy of the aging human heart. Myocyte loss and reactive cellular hypertrophy. Circ. Res. 1991, 68, 1560–1568.
    36. Kajstura, J.; Cheng, W.; Sarangarajan, R.; Li, P.; Li, B.; Nitahara, J.A.; Chapnick, S.; Reiss, K.; Olivetti, G.; Anversa, P. Necrotic and apoptotic myocyte cell death in the aging heart of Fischer 344 rats. Am. J. Physiol. 1996, 271, H1215–H1228.
    37. Kung, G.; Konstantinidis, K.; Kitsis, R.N. Programmed necrosis, not apoptosis, in the heart. Circ. Res. 2011, 108, 1017–1036.
    38. Zhang, X.; Wu, J.Z.; Lin, Z.X.; Yuan, Q.J.; Li, Y.C.; Liang, J.L.; Zhan, J.Y.X.; Xie, Y.L.; Su, Z.R.; Liu, Y.H. Ameliorative effect of supercritical fluid extract of Chrysanthemum indicum Linnén against D-galactose induced brain and liver injury in senescent mice via suppression of oxidative stress, inflammation and apoptosis. J. Ethnopharmacol. 2019, 234, 44–56.
    39. Ullah, F.; Ali, T.; Ullah, N.; Kim, M.O. Caffeine prevents d-galactose-induced cognitive deficits, oxidative stress, neuroinflammation and neurodegeneration in the adult rat brain. Neurochem. Int. 2015, 90, 114–124.
    40. Sun, S.L.; Guo, L.; Ren, Y.C.; Wang, B.; Li, R.H.; Qi, Y.S.; Yu, H.; Chang, N.D.; Li, M.H.; Peng, H.S. Anti-apoptosis effect of polysaccharide isolated from the seeds of Cuscuta chinensis Lam on cardiomyocytes in aging rats. Mol. Biol. Rep. 2014, 41, 6117–6124.
    41. Abulfadl, Y.S.; El-Maraghy, N.N.; Ahmed, A.A.E.; Nofal, S.; Badary, O.A. Protective effects of thymoquinone on D-galactose and aluminum chloride induced neurotoxicity in rats: Biochemical, histological and behavioral changes. Neurol. Res. 2018, 40, 324–333.
    42. Banji, D.; Banji, O.J.F.F.; Dasaroju, S.; Kumar Ch, K. Curcumin and piperine abrogate lipid and protein oxidation induced by d-galactose in rat brain. Brain Res. 2013, 1515, 1–11.
    43. Banji, O.J.F.F.; Banji, D.; Ch, K. Curcumin and hesperidin improve cognition by suppressing mitochondrial dysfunction and apoptosis induced by D-galactose in rat brain. Food Chem. Toxicol. 2014, 74, 51–59.
    44. Fairless, R.; Williams, S.K.; Diem, R. Dysfunction of neuronal calcium signalling in neuroinflammation and neurodegeneration. Cell Tissue Res. 2014, 357, 455–462.
    45. Fairless, R.; Williams, S.K.; Diem, R. Calcium-Binding Proteins as Determinants of Central Nervous System Neuronal Vulnerability to Disease. Int. J. Mol. Sci. 2019, 20, 2146.
    46. Ouh, I.-O.; Kim, Y.-M.; Gim, S.-A.; Koh, P.-O. Focal cerebral ischemic injury decreases calbindin expression in brain tissue and HT22 cells. Lab. Anim. Res. 2013, 29, 156.
    47. Tai, Y.H.; Lin, Y.Y.; Wang, K.C.; Chang, C.L.; Chen, R.Y.; Wu, C.C.; Cheng, I.H. Curcuminoid submicron particle ameliorates cognitive deficits and decreases amyloid pathology in Alzheimer’s disease mouse model. Oncotarget 2018, 9, 10681–10697.
    48. Sasaki, Y.; Ohsawa, K.; Kanazawa, H.; Kohsaka, S.; Imai, Y. Iba1 is an actin-cross-linking protein in macrophages/microglia. Biochem. Biophys. Res. Commun. 2001, 286, 292–297.
    49. Ito, D.; Tanaka, K.; Suzuki, S.; Dembo, T.; Fukuuchi, Y. Enhanced Expression of Iba1, Ionized Calcium-Binding Adapter Molecule 1, After Transient Focal Cerebral Ischemia In Rat Brain. Stroke 2001, 32, 1208–1215.
    50. Ton, B.H.T.; Chen, Q.; Gaina, G.; Tucureanu, C.; Georgescu, A.; Strungaru, C.; Flonta, M.L.; Sah, D.; Ristoiu, V. Activation profile of dorsal root ganglia Iba-1 (+) macrophages varies with the type of lesion in rats. Acta Histochem. 2013, 115, 840–850.
    51. Romero-Sandoval, A.; Chai, N.; Nutile-McMenemy, N.; DeLeo, J.A. A comparison of spinal Iba1 and GFAP expression in rodent models of acute and chronic pain. Brain Res. 2008, 1219, 116–126.
    52. Eitzen, U.V.; Egensperger, R.; Kösel, S.; Grasbon-Frodl, E.M.; Imai, Y.; Bise, K.; Kohsaka, S.; Mehraein, P.; Graeber, M.B. Microglia and the Development of Spongiform Change in Creutzfeldt-Jakob Disease. J. Neuropathol. Exp. Neurol. 1998, 57, 246–256.
    53. Ullah, R.; Jo, M.H.; Riaz, M.; Alam, S.I.; Saeed, K.; Ali, W.; Rehman, I.U.; Ikram, M.; Kim, M.O. Glycine, the smallest amino acid, confers neuroprotection against D-galactose-induced neurodegeneration and memory impairment by regulating c-Jun N-terminal kinase in the mouse brain. J. Neuroinflamm. 2020, 17, 303.
    54. Duan, S.; Wang, X.; Chen, G.; Quan, C.; Qu, S.; Tong, J. Inhibiting RIPK1 Limits Neuroinflammation and Alleviates Postoperative Cognitive Impairments in D-Galactose-Induced Aged Mice. Front. Behav. Neurosci. 2018, 12, 138.