Li Juan, Tian Xiangge, Liu Jinming, Mo Yuying, Guo Xiaoyi, Qiu Yang, Liu Yuejian, Ma Xiaochi, Wang Yan, Xiong Yongjian
Central Laboratory, First Affiliated Hospital of Dalian Medical University, Dalian, 116011, China.
Department of Pharmacy, Peking University Shenzhen Hospital, Shenzhen, 518036, China.
Chin Med. 2022 Aug 16;17(1):96. doi: 10.1186/s13020-022-00652-1.
Shaoyao Decoction (SYD) is a canonical herbal medicine prescription formulated by Liu Wan-Su in AD 1186. SYD has been widely used to treat inflammatory bowel disease by clearing heat and damp, removing stasis toxin in the intestine; however, the precise mechanisms and therapeutic material basis remain largely unclear. In the present study, we measured the effects of SYD on colitis symptom, epithelial barrier function, epithelial ferroptosis, colonic protein and mRNA expression of glutathione peroxidase 4 (GPX4) in colitis model, and determined whether SYD restored barrier loss in colitis by modulation of GPX4-regulated ferroptosis pathway.
Colitis was established by infusion with 1 mL 2,4,6-trinitrobenzene sulfonic acid (TNBS) dissolved in ethanol (40% v/v) in rats at a 125 mg/kg dose. Ferroptosis in epithelial cells was determined by flow cytometer. GPX4 promoter-firefly luciferase fusion construct was transfected to Caco-2 cell to determine GPX4 transcription. MS analysis was used to identified ingredients in SYD.
Different doses of SYD significantly alleviated colitis, decreased ferroptosis in epithelial cells, knockout of GPX4 significantly reversed SYD-induced alleviation effects on colitis, restoration of epithelial barrier function, and epithelial ferroptosis. Wogonoside, wogonin, palmatine, paeoniflorin and liquiritin were identified as active ingredients of SYD-exerted alleviation effects of colitis based on GPX4 agonistic transcription.
SYD alleviated chemically induced colitis by activation of GPX4, inhibition of ferroptosis in epithelial cells and further restoration of barrier function. Wogonoside, wogonin, palmatine, paeoniflorin and liquiritin were identified as the key therapeutic material basis of SYD-exerted anti-colitis effects. The findings provide a scientific basis for the therapeutic effect of SYD on colitis.
芍药汤（SYD）是刘完素于公元1186年制定的经典中药方剂。芍药汤已被广泛用于通过清热利湿、清除肠道内的瘀血毒素来治疗炎症性肠病；然而，其确切机制和治疗物质基础仍不清楚。在本研究中，我们测量了芍药汤对结肠炎模型中结肠炎症状、上皮屏障功能、上皮铁死亡、结肠中谷胱甘肽过氧化物酶4（GPX4）的蛋白质和mRNA表达的影响，并确定芍药汤是否通过调节GPX4调节的铁死亡途径恢复结肠炎中的屏障损失。
通过以125mg/kg的剂量向大鼠输注1mL溶解于乙醇（40%v/v）中的2,4,6-三硝基苯磺酸（TNBS）来建立结肠炎模型。通过流式细胞仪测定上皮细胞中的铁死亡。将GPX4启动子-萤火虫荧光素酶融合构建体转染至Caco-2细胞以确定GPX4转录。采用质谱分析来鉴定芍药汤中的成分。
不同剂量的芍药汤显著减轻结肠炎，降低上皮细胞中的铁死亡，敲除GPX4显著逆转芍药汤诱导的对结肠炎的缓解作用、上皮屏障功能的恢复以及上皮铁死亡。基于GPX4激动转录，确定汉黄芩苷、汉黄芩素、巴马汀、芍药苷和甘草苷为芍药汤发挥结肠炎缓解作用的活性成分。
芍药汤通过激活GPX4、抑制上皮细胞中的铁死亡并进一步恢复屏障功能来减轻化学诱导的结肠炎。汉黄芩苷、汉黄芩素、巴马汀、芍药苷和甘草苷被确定为芍药汤发挥抗结肠炎作用的关键治疗物质基础。这些发现为芍药汤治疗结肠炎的疗效提供了科学依据。
