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炎调方调控Rho/ROCK信号通路对脓毒症急性胃肠损伤小鼠肠上皮细胞凋亡的研究OACSTPCD

Mechanism of Yantiao Fang regulating the Rho/ROCK signaling pathway in apoptosis of intestinal epithelial cells in mice with acute gastrointestinal injury caused by sepsis

中文摘要英文摘要

目的 探讨炎调方调控Rho/ROCK信号通路对脓毒症急性胃肠损伤小鼠肠上皮细胞凋亡的影响.方法 取70只BALB/c小鼠随机分为空白组、假手术组和造模小鼠.除空白组、假手术组外,各组小鼠均采用盲肠结扎穿孔术(cecum ligation and puncture,CLP)构建脓毒症急性胃肠损伤小鼠模型,将造模成功的小鼠随机分为模型组、炎调方低、中、高剂量组和ROCK抑制剂组.采用HE染色观察小鼠回肠组织病理学改变;ELISA法检测各组小鼠血清IL-1β、IL-6、TNF-α、IL-10水平;免疫组织化学检测观察PCNA、Ki-67的表达;蛋白印迹法检测回肠组织凋亡指标cleaved caspase3蛋白、Bax蛋白表达;RT-qPCR检测小鼠回肠组织ROCK mRNA、MLC mRNA表达.结果 较空白组和假手术组相比,模型组小鼠的Chiu's病理评分、促炎介质(IL-1β、IL-6、TNF-α)水平、cleaved caspase3蛋白、Bax蛋白、ROCKmRNA、MLC mRNA水平升高(P<0.05),而抑炎介质(IL-10)水平和回肠组织中PCNA、Ki-67表达降低(P<0.05).与模型组比较,炎调方各组随着剂量增加,回肠组织病理学改变均得到不同程度改善,其Chiu's 病理评分、血清中 IL-1β、IL-6、TNF-α 水平、cleaved caspase3 蛋白、Bax 蛋白、ROCK mRNA、MLC mRNA 的表达降低(P<0.05),而IL-10水平、回肠组织中PCNA、Ki-67表达升高(P<0.05).结论 炎调方可能通过调控Rho/ROCK信号通路抑制脓毒症急性胃肠损伤小鼠的肠上皮细胞凋亡,从而具有减轻肠道组织炎症反应,最终达到防止肠黏膜组织损伤的作用.

Objective To investigate the effect of Yantiao Fang on apoptosis of intestinal epithelial cells in mice with acute gastrointestinal injury caused by sepsis by regulating the Rho/ROCK signaling pathway.Methods Seventy BALB/c mice were randomly divided into normal,sham,and model groups.Except for normal and sham groups,mice were subjected to cecal ligation and perforation to establish a mouse model of acute gastrointestinal injury caused by sepsis.The mouse models were randomly divided into model,Low,Medium,and High dose of Yantiao Fang,and ROCK inhibitor groups.Histopathological changes of the ileum were observed by HE staining.Serum levels of IL-1β,IL-6,TNF-α,and IL-10 were measured by ELISA.PCNA and Ki-67 expression was detected by immunohistochemistry.Cleaved caspase3 and Bax expression was detected by Western blot.ROCK and MLC mRNA expression in the ileum was measured by RT-qPCR.Results Compared with normal and sham groups,Chiu's pathological score,proinflammatory factor(IL-1β,IL-6,and TNF-α)levels,cleaved caspase3 and Bax protein expression,and ROCK and MLC mRNA levels were increased in the model group(P<0.05).Moreover,anti-inflammatory mediator IL-10 and expression of PCNA and Ki-67 in the ileum were decreased in the model group(P<0.05).Compared with the model group,histopathological changes of the ileum in all Yantiao Fang groups were improved by various degrees with the increase in dose.Chiu's pathological score,IL-1β,IL-6,and TNF-α serum levels,cleaved caspase3 and Bax protein expression,and ROCK and MLC mRNA levels were decreased in Yantian Fang groups(P<0.05).The IL-10 level and expression of PCNA and Ki-67 in the ileum were increased in Yantian Fang groups(P<0.05).Conclusions Yantiao Fang may inhibit apoptosis of intestinal epithelial cells in mice with acute gastrointestinal injury due to sepsis by regulating the Rho/ROCK signaling pathway,thereby alleviating intestinal tissue inflammation and ultimately preventing intestinal mucosal tissue injury.

陈乾;王帝;张定一;贾赛蕾;王丽辉;曹玉净;李鲜;李燕红

河南省中医院/河南中医药大学第二附属医院,郑州 450002||黄河科技学院,郑州 450000河南省中医院/河南中医药大学第二附属医院,郑州 450002黄河科技学院,郑州 450000

脓毒症急性胃肠损伤肠上皮细胞凋亡Rho/ROCK信号通路炎调方

acute gastrointestinal injury caused by sepsisapoptosis of intestinal epithelial cellsRho/ROCK signaling pathwayYantiao Fang

《中国比较医学杂志》 2024 (002)

16-24 / 9

国家自然科学基金青年项目(82104833);全国名老中医药专家传承工作室(国中医药人教函[2022]75号);河南省中医药科学研究专项课题(2024ZY2084,20-21ZY2114,2022ZY1086);河南省科技攻关项目(212102310339).

10.3969/j.issn.1671-7856.2024.02.003

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