背景:五味子木脂素(SCL),传统功能性中药五味子的主要活性成分,据报道有抗抑郁作用。其机制包括通过解决肠道微生物区系来减轻肠道屏障损伤(IBI),抗炎,和神经保护。SCL还调节内源性大麻素系统,与抑郁症的发生发展密切相关。
目的:我们研究了一种新的抑郁症治疗策略,即,通过调节内源性大麻素系统的抗抑郁作用来缓解IBI,并对其具体机制进行了深入的研究探讨。
方法:进行行为分析以检测C57BL/6小鼠中抑郁样行为的发生。我们用苏木精-伊红染色,高碘酸希夫染色,和免疫荧光来评估IBI。网络药理学和Westernblotting(WB)用于预测和证实SCL的改善作用与抗炎和抗凋亡有关。结合Anandamide(AEA)和2-花生四酰基甘油(2-AG)的水平,我们在AEA之间进行了Pearson分析,2-AG水平以及通过网络药理学和WB鉴定和验证的主要靶标。随后,URB-597,一种具有AEA水解酶抑制作用的脂肪酸酰胺水解酶(FAAH)拮抗剂,给老鼠服用,并对行为分析和凋亡蛋白进行了验证。通过6470三重四极杆LC/MS测量补充URB-597后的血浆内源性大麻素水平。最后,将大麻素受体2型(CB2R)拮抗剂AM630给予小鼠,并进行免疫荧光和WB以评估IBI和抗炎的蛋白质。
结果:研究表明,SCL减轻了抑郁样行为并改善了IBI。网络药理学和WB证实IBI的改善与抗炎和抗凋亡途径有关。Pearson结果显示,AEA水平与炎症和细胞凋亡呈正相关,对细胞凋亡的贡献更大。深入研究验证了URB-597的给药逆转了SCL对抑郁样行为和抗凋亡的积极作用。同样,URB-597抵消了SCL降低的AEA水平,并降低了2-AG水平。此外,补充AM630拮抗SCL通过重新激活MAPK/NF-κB炎症通路改善IBI的作用。
结论:总体而言,SCL,与SCL调节的内源性大麻素系统合作,缓解IBI相关的抑郁症。具体机制涉及SCL降低AEA水平以通过上调FAAH来抑制结肠组织细胞凋亡。同时,它直接触发CB2R以减少炎症反应,进一步缓解IBI。
BACKGROUND: Schisandra chinensis lignan (SCL), a major active component of the traditional functional Chinese medicine Schisandra chinensis, has been reported to have antidepressant effects. Its mechanisms include alleviating intestinal barrier injury (IBI) by resolving intestinal microflora, anti-inflammation, and neuroprotection. SCL also regulates endogenous cannabinoid system, and it is closely related to the onset and development of depression.
OBJECTIVE: We investigated a new treatment strategy for depression, i.e., alleviating IBI by regulating the endogenous cannabinoid system for antidepressant effects, as well as conducted in-depth research to explore the specific mechanism.
METHODS: Behavioral analysis was conducted to detect the occurrence of depressive-like behavior in C57BL/6 mice. We used hematoxylin-eosin staining, periodic acid-Schiff staining, and immunofluorescence to evaluate IBI. Network pharmacology and Western blotting (WB) were used to predict and confirm that the amelioration effect of SCL was associated with anti-inflammation and anti-apoptosis. Combined with the levels of anandamide (AEA) and 2-arachidonoylglycerol (2-AG), we conducted the Pearson analysis between the AEA, 2-AG levels and the major targets identified and validated by network pharmacology and WB. Subsequently, URB-597, a fatty acid amide hydrolase (FAAH) antagonist with an AEA hydrolase-inhibiting effect, was administered to the mice, and behavioral analysis and apoptotic proteins were verified. Plasma endocannabinoid levels after URB-597 supplementation were measured via 6470 Triple Quadrupole LC/MS. Finally, the cannabinoid receptor type 2 (CB2R) antagonist AM630 was administered to mice, and immunofluorescence and WB were performed to assess the proteins of IBI and anti-inflammation.
RESULTS: The study demonstrated that SCL alleviated depressive-like behaviours and ameliorated IBI. Network pharmacology and WB confirmed that the improvement of IBI was related to the anti-inflammatory and anti-apoptotic pathways. Pearson results showed that AEA levels were positively correlated with inflammation and apoptosis, with a greater contribution to apoptosis. In-depth studies validated that the URB-597 administration reversed the positive effects of SCL on depressive-like behavior and anti-apoptosis. Similarly, URB-597 counteracted AEA levels reduced by SCL and decreased 2-AG levels. Furthermore, AM630 supplementation antagonized SCL\'s effect of improving IBI by reactivating the MAPK/NF-κB inflammation pathway.
CONCLUSIONS: Overall, SCL, in collaboration with the endogenous cannabinoid system regulated by SCL, alleviates depression associated IBI. The specific mechanism involes SCL decreasing AEA levels to inhibit colon tissue cell apoptosis by up-regulating FAAH. Simultaneously, it directly triggers CB2R to reduce inflammation responses, further alleviating IBI.