Chrysin (
Figure 1) has a backbone structure that consists of a fused A and C rings, and a phenyl B ring, which is attached to the second position of ring C and shares the basic structure of the flavones, with an additional hydroxyl group at the fifth and seventh positions of the A ring. The potential of chrysin to act as a free radical scavenger has been attributed to the presence of these hydroxyl groups
[1][2], and it has been suggested that these functional groups represent the main site of action of this flavonoid to produce a great variety of pharmacological activities and therapeutic effects. It has the potential to be used as an alternative in the treatment of metabolic, cardiovascular, and neuropsychiatric disorders
[3]. In addition, the presence of hydroxyl groups in the backbone of chrysin has been associated with its anxiolytic-like effects
[4]. Chrysin, but not the flavone backbone, decreases anxiety-like behavior in rats and zebrafish, suggesting that the presence of hydroxyl groups in its basic structure is indispensable for producing anxiolytic-like effects in pre-clinical research
[4].
2. Anxiolytic-like Effects of Flavonoid Chrysin
In 1994, Wolfman et al. reported an anxiolytic-like effect of chrysin in mice. A single dose of chrysin at 1 mg/kg significantly increased the time spent in open arms of the EPM
[9]. In the light/dark box (LDB), increased time was spent in the illuminated compartment
[7], and in both cases the effects were similar to that produced by diazepam. These behavioral effects are considered to be associated with anxiolytic-like effects in pre-clinical research. Chrysin, but not diazepam, is devoid of motor effects related to sedation
[9] and this may represent the advantage of chrysin over benzodiazepines, such as diazepam, in the treatment of anxiety disorders
[12]. Interestingly, in male Sprague Dawley rats, the anxiolytic-like effects of chrysin at 1 and 2 mg/kg in LDB were blocked by a previous administration of flumazenil
[7][13], an antagonist of the benzodiazepine binding site in the GABA
A receptor. Additionally, acute administration of chrysin (1 mg/kg) produced anxiolytic-like effects in male Wistar rats
[14] and CD-1 male mice
[15] evaluated in the EPM. Similarly, anxiolytic-like effects of chrysin in mammals (mice and rats) have also been reported in non-mammalian organisms (zebrafish). Chrysin at 1 mg/kg decreased anxiety-like behavior in rats and zebrafish, similar to diazepam
[4]; however, treatment with a flavone backbone at 1 mg/kg was devoid of anxiolytic-like effects in both rats and zebrafish, suggesting that the presence of hydroxyl groups in its basic structure could be indispensable to produce anxiolytic-like effects
[4].
Anxiety symptoms in women are associated with a reduction in steroid hormones, such as estradiol and progesterone, and its reduced metabolite allopregnanolone in the peripheral and CNS, which may occur pre-menstruation, post-partum, and during the transition to menopause stage
[16][17]. These steroid hormones may modulate several neurotransmission systems, such as the serotoninergic, noradrenergic, dopaminergic, and GABAergic
[18]; therefore, some of these hormones have been proposed as novel groups of anxiolytic drugs for treating particular anxiety and depression disorders associated with reduced concentrations of steroid hormones
[19]. It has recently been proposed that the flavonoid chrysin mimics some of the pharmacological effects of neurosteroids in female rats
[20]. Anxiety-like behaviors in female rats significantly increase during the metestrus–diestrus phase of the ovarian cycle, which is associated with a low concentration of steroid hormones
[21]; this phase is considered an equivalent of the premenstrual period in women
[22]. Interestingly, chrysin at 2 mg/kg, similar to diazepam at 2 mg/kg, prevents anxiety-like behavior that naturally occurs during the metestrus–diestrus phase in female rats evaluated in the EPM and LDB. This effect can be blocked by a previous injection of picrotoxin
[23]. In support, microinjection of chrysin at 0.5 μg in the dorsal HP prevented anxiety-like behavior that naturally occurs during diestrus, which was blocked by previous injection of picrotoxin, bicuculline, and flumazenil, indicating that the GABA/benzodiazepine receptor complex in the dorsal HP mediates the anxiolytic-like effects of this flavonoid
[24]. Interestingly, this same effect on anxiety-like behavior during diestrus was prevented by microinjection of neurosteroid allopregnanolone at 0.5 μg into the dorsal HP, which was blocked by picrotoxin, bicuculline, and flumazenil in the EPM
[24]. In contrast, in a surgical menopause model in rats characterized by high anxiety-like behavior associated with a permanent reduction of steroid hormones, chrysin at 2 and 4 mg/kg and diazepam at 1 mg/kg, reversed this anxiety-like behavior, which was blocked by a previous injection of picrotoxin
[11]. The fact that picrotoxin, bicuculline, and flumazenil prevented the anxiolytic-like effect of different doses of chrysin supports the idea that its pharmacological effects are established on the GABA/benzodiazepine receptor complex, as occurs with clinically effective GABAergic anxiolytic drugs and several neurosteroids, such as allopregnanolone
[25], but does not produce the typical sedative effects of benzodiazepines
[9]. However, the researchers cannot discard the possibility of other neurotransmitter systems’ participation and the anti-inflammatory and antioxidant effects in different regions of the brain due to the anxiolytic-like effects of chrysin (
Figure 2). Specific studies are required to support or discard this possibility.
Figure 2. Mechanism of action of the flavonoid chrysin potentially involved in its anxiolytic-like effects. (
A) It has been confirmed that chrysin produces its anxiolytic-like effect through its action on the GABA
A/benzodiazepine receptor complex producing configurational changes in the receptor and regulating the opening of the Cl
− ion channel
[5][7][9][26][27], which may produce inhibitory effects in the GABAergic system associated with its anxiolytic-like effects. These effects can be blocked by specific antagonists of the GABA
A receptor, such as picrotoxin, bicuculline, and flumazenil
[5]. (
B) Probably, antioxidant effects of chrysin could be involved in its anxiolytic-like effects. Chrysin significantly reduces ROS by inhibiting the production of NO, NT, and NOX4
[28]. These effects reduce the oxidative stress and reduces the neuronal damage. Additionally, chrysin reduces the activity of Bax, caspase-9, and caspase-3, while increasing the production of Bcl-2, thereby reducing the damage of DNA and inhibiting apoptotic processes
[29][30], which reduces the neuronal death. (
C) Additionally, the anti-inflammatory effects of chrysin could contribute to its anxiolytic-like effects, considering that it may reduce the inflammatory response by inhibiting the signaling pathway NF-κB/IKK-β
[31][32]. Chrysin may attenuate the expression of NF-κB that participates as transcriptional factors at nuclear level, binding to genes that induce neuro-inflammation process. Chrysin also inhibits the production of pro-inflammatory cytokines, such as IL-1β and IL-6, in addition to suppressing the production of proinflammatory mediators, such as TNF-α, PGE
2 and COX-2
[31][32][33]. These effects could reduce neuro-inflammation associated with the anxiety-like behavior. ROS = reactive oxygen species; NO = nitric oxide; NT = nitrotyrosine; NOX4 = NADPH oxidase; O2¯ = superoxide; Green circles = chlorine ions; SOD = superoxide dismutase; GSH = reduced glutathione; CAT = catalase; GPx = glutathione peroxidase; Bcl-2 = anti-apoptotic protein of the subfamily Bcl-2; Bax = pro-apoptotic protein of the subfamily Bax; NF-κB = nuclear factor kappa B; IKK-β = inhibitor of nuclear factor kappa-B; TNF-α = tumor necrosis factor-α; IL-1β = interleukin-1β; IL-6 = interleukin-6; PGE2 = prostaglandins E2; COX-2 = cycloxygenase-2. (Figure was prepared by the authors).
3. Antidepressant-like Effects of Flavonoid Chrysin
Few studies have explored the antidepressant-like effects of chrysin; however, their results are promising. Filho et al.
[8] reported that chrysin at 5 and 20 mg/kg for 28 days increased sucrose consumption and decreased immobility in the tail suspension test (TST) in female mice C57B/6J exposed to CUMS, which is considered to have antidepressant-like effects in pre-clinical research. This effect was also associated with an increase in serotonin, BDNF, and NGF levels, and decreased pro-inflammatory levels of cytokines, such as TNF-α, IFN-γ, IL-1β, and IL-6 in the HP and PFC of C57B/6J mice
[8][34]. Additionally, chrysin at 20 mg/kg for 14 days produced an antidepressant-like effect in the FST in male mice C57B/6J subjected to depression induced by olfactory bulbectomy. This effect was associated with decreased pro-inflammatory cytokines (i.e., TNF-α, IFN-γ, IL-1β, IL-6), kynurenine (KYN, a metabolite resulting from serotonin degradation), and indolamine-2, 3-dyoxigenase (IDO, enzyme responsible for serotonin metabolism) activity, besides producing an increase in BDNF and serotonin levels in HP
[10]. Interestingly, chrysin at 1, 5, and 10 mg/kg for 28 days produced antidepressant-like effects in the FST in male Wistar rats
[35]. In addition, chrysin at 1 and 5 mg/kg for 28 days significantly reduced 5-HT
1A receptor expression in the raphe nucleus and increased it in HP, whereas 5-HT
2A receptor expression was increased in HP
[35]. These effects were similar to those produced by the antidepressant fluoxetine at 1 mg/kg for 28 days. In another study, chrysin at 20 mg/kg for 28 days produced antidepressant-like effects in the TST and FST in female C57BL/6 mice exposed to a depression model induced by hypothyroidism, which was associated with increased serotonin and dopamine levels in the HP
[36].
As previously mentioned, a reduced concentration of ovarian hormones in women during their transition to menopause, increases the risk of developing anxiety and depression symptoms
[37]. Interestingly, using a surgical menopause model in Wistar rats, it was reported that chrysin at 1 mg/kg reversed depression-like behavior in the FST; this effect was similar to that produced by neurosteroids progesterone at 1 mg/kg and allopregnanolone at 1 mg/kg
[20]. The effects of chrysin and neurosteroids were blocked by the previous administration of bicuculline, a selective competitive antagonist of the binding site of γ-aminobutyric acid in the GABA
A receptor, which supports the idea that activation of the GABAergic system participates in the antidepressant-like effect of chrysin, as has been reported with neurosteroids
[38].
Based on the results described above, the researchers suggest that the mechanism of action underlying the antidepressant-like effect of chrysin involves multiple neurochemical processes, such as the activation of neurotransmitter systems, anti-inflammatory and antioxidant processes, and the activation of neurotrophic factors (Figure 3); however, further exploration is required to improve people's understanding of these mechanisms underlying the antidepressant-like effects of chrysin, and to explore its effects in controlled clinical trials.
Figure 3. Possible mechanisms of action involved in the antidepressant-like effect of chrysin. (
A) The flavonoid chrysin can modulate ERα and ERβ of membrane, which triggers the MAPK/ERK1/2 signaling pathway involved in phosphorylation and subsequent CREB activation (
↑CREB), which promotes the increase of BDNF levels (
↑BDNF)
[1][34][39][40], which can further activate the MAPK/ERK1/2 signaling by the TrkB interaction
[41]. The above-mentioned pathway also promotes an increase of TpOH expression (
↑TpOH) and serotonin levels (
↑Serotonin) resulting in the antidepressant-like effect
[10][36]. (
B) Furthermore, chrysin can decrease the pro-inflammatory cytokine levels (
↓TNF-α,
↓IL-1β,
↓IL-6,
↓IFN-γ), which inhibits IDO activity (
↓IDO) improving serotonergic neurotransmission and producing its antidepressant-like effect
[10]. ER = estrogen receptor; MAPK = mitogen-activated-protein-kinases; CREB = cAMP response element binding; BDNF = brain derived neurotrophic factor; TrkB = tropomyosinreceptor kinase B; TpOH = tryptophan-hydroxylase; TNF-α = tumor necrosis factor-α; IL-1β = interleukin 1 beta; IL-6 = interleukin 6; IFN-γ = interferon gamma; IDO = indoleamine 2,3-dioxygenase; ERK1/2 = extracellular signal-regulated kinase 1 and 2; KYN = kynurenine. (Figure was prepared by the authors).