Zhang Xiao-Yun, Zhao Hai-Mei, Liu Yi, Lu Xiu-Yun, Li Yan-Zhen, Pan Qi-Hong, Wang Hai-Yan, Ge Wei, Liu Duan-Yong
Department of Postgraduate, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, Jiangxi, China.
School of Basic Medical Sciences, Jiangxi University of Traditional Chinese Medicine, Nanchang 330004, Jiangxi, China.
Evid Based Complement Alternat Med. 2021 Dec 9;2021:5536679. doi: 10.1155/2021/5536679. eCollection 2021.
Sishen Pill (SSP) is a classical prescription of traditional Chinese medicine and often used to treat gastrointestinal diseases, including ulcerative colitis (UC). However, its mechanism is still unclear. We aimed to determine the mechanism of SSP in the treatment of UC by investigating if it maintains the integrity of the intestinal mucosal barrier via the Rho A/Rho kinase (ROCK) signaling pathway. Administration of 2,4,6-trinitrobenzene sulfonic acid (TNBS) successfully induced chronic UC in rats, while the treatment effect of SSP was evaluated by body weight change, colonic length, colonic weight, colonic weight index, histological injury score, and pathological injury score after colitis rats were treated for 7 days. TNF- and IL-1 levels were analyzed by ELISA, and the proteins of PI3K/Akt and RhoA/ROCK signaling pathway and junction proteins expression were measured by western blotting assay, and the distribution of Claudin 5 was shown by immunofluorescence. SSP significantly improved the clinical symptoms of colitis in rats and reduced the expression of p-RhoA, ROCK1, PI3K, and Akt in the colon mucosa, while it increased the expression of p-Rac and related proteins (Claudin-5, JAM1, VE-cadherin, and Connexin 43). In addition, SSP increased p-AMPK and PTEN proteins expression, decreased Notch1 level, and hinted that activation of the PI3K/Akt signaling pathway was inhibited. In conclusion, SSP effectively treated chronic colitis induced by TNBS, which may have been achieved by inhibiting PI3K/Akt signal to suppress activation of the Rho/ROCK signaling pathway to finally maintain the integrity of the intestinal mucosal barrier.
四神丸（SSP）是一种经典的中药方剂，常用于治疗胃肠道疾病，包括溃疡性结肠炎（UC）。然而，其作用机制仍不清楚。我们旨在通过研究SSP是否通过Rho A/ Rho激酶（ROCK）信号通路维持肠道黏膜屏障的完整性，来确定其治疗UC的机制。给予2,4,6-三硝基苯磺酸（TNBS）成功诱导大鼠慢性UC，在对结肠炎大鼠治疗7天后，通过体重变化、结肠长度、结肠重量、结肠重量指数、组织学损伤评分和病理损伤评分来评估SSP的治疗效果。通过酶联免疫吸附测定（ELISA）分析肿瘤坏死因子（TNF）和白细胞介素-1（IL-1）水平，采用蛋白质免疫印迹法检测PI3K/Akt和RhoA/ROCK信号通路相关蛋白以及紧密连接蛋白的表达，并通过免疫荧光显示Claudin 5的分布。SSP显著改善了大鼠结肠炎的临床症状，降低了结肠黏膜中p-RhoA、ROCK1、PI3K和Akt的表达，同时增加了p-Rac及相关蛋白（Claudin-5、连接黏附分子1、血管内皮钙黏蛋白和连接蛋白43）的表达。此外，SSP增加了p-AMPK和PTEN蛋白的表达，降低了Notch1水平，提示PI3K/Akt信号通路的激活受到抑制。总之，SSP有效治疗了TNBS诱导的慢性结肠炎，这可能是通过抑制PI3K/Akt信号来抑制Rho/ROCK信号通路的激活，最终维持肠道黏膜屏障的完整性来实现的。
