基于MLCK/MLC信号通路探讨四逆散对溃疡性结肠炎小鼠肠上皮屏障的调节作用  

Regulation of Sini Powder(四逆散)of the Intestinal Epithelial Barrier by Targeting the MLCK/MLC Signaling Pathway in Mouse Model of Experimental Colitis

作　　者：魏新智[1,2] 樊晖 于艳[1] 鞠业涛[1] WEI Xinzhi;FAN Hui;YU Yan;JU Yetao(Afliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China;Shenyang Sports Universtity,Shenyang 110101,Liaoning,China)

机构地区：[1]辽宁中医药大学附属医院,辽宁沈阳110032 [2]沈阳体育学院,辽宁沈阳110101

出　　处：《辽宁中医药大学学报》2025年第4期28-33,共6页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：辽宁省教育厅资助项目(L202017)。

摘　　要：目的基于MLCK/MLC信号探讨四逆散对溃疡性结肠炎小鼠结肠上皮屏障的调节作用。方法C57BL/6小鼠按体质量随机分为对照组(CON),模型组(TNBS),四逆散低(SND-L)、高(SND-H)剂量组,采用三硝基苯磺酸(2,4,6-trinitro-benzene-sulfonic acid,TNBS)与无水乙醇1∶1比例混合液灌肠建立小鼠溃疡性结肠炎模型,比较各组小鼠体质量变化;FITC-右旋糖酐渗透试验检测小鼠结肠通透性;HE染色观察结肠的组织学变化;实时荧光定量PCR检测小鼠结肠黏膜中的细胞因子TNFA、IL-1b、IL-6、IL-17 mRNA的表达;Western blotting检测小鼠结肠黏膜中MLCK、p-MLC、occludin、PUMA蛋白的表达。结果与CON组相比,TNBS组小鼠体质量下降,结肠变短;小鼠结肠管腔粗细不均,粪便不成形或集结成块状,伴有充血或出血;组织学观察小鼠结肠黏膜完整性破坏、炎细胞浸润,大小不等的溃疡面,部分腺体消失;小鼠结肠FITC-右旋糖酐渗透性增加;TNFA、IL-1b、IL-6、IL-17 mRNA表达升高;MLCK、p-MLC、PUMA蛋白表达量显著升高,occludin蛋白的表达下降。与TNBS组相比,四逆散组小鼠体质量下降比例减小、结肠变长;小鼠结肠FITC-右旋糖酐渗透性降低;小鼠结肠黏膜炎细胞浸润减轻,部分腺体恢复,溃疡面积减小;小鼠结肠黏膜中TNFA、IL-1b、IL-6、IL-17 mRNA表达降低;MLCK、p-MLC、PUMA蛋白表达量下降,occludin蛋白的表达升高。结论四逆散有减轻小鼠溃疡性结肠炎的作用,并可认为是与抑制小鼠结肠黏膜中MLCK、p-MLC、PUMA蛋白表达,升高occludin蛋白的表达,改善小鼠肠屏障功能相关。Objective To investigate whether Sini Powder(四逆散,SND)suppresses the MLCK/MLC signaling pathway to prevent colonic mucosal infammation in a mouse model of experimental colitis.Methods C57BL/6 mice were randomly divided into control group(CON),model group(TNBS),lowdose group(SND-L)and high-dose group(SND-H)of Sinisan.C57BL/6 mice were induced to colitis by 2,4,6-trinitrobenzenesulfonic acid(TNBS)to evaluate the effect of SND.The general behavior and body weight after modeling was observed and recorded.The weight loss index and the gross score of the colon in each group were compared.FITC-dextran permeability test was used to detect colonic mucosal permeability.HE staining was used to observe the pathological changes of colon tissue and pathological scores were performed.The expression of inflammatory cytokines TNFA,IL-1b,IL-6 and IL-17 mRNA were quantifed in mouse colon mucosa by Real-time PCR.The expression of MLCK,p-MLC,occludin and PUMA proteins in colonic mucosa of mice by Western blotting.Results Compared to the CON group,the TNBS group exhibited decreased body weight,shortened colons,uneven colon lumen diameter,loose or clumped stools accompanied by congestion or hemorrhage.Histological observations revealed disrupted colonic mucosal integrity,inflammatory cell infiltration,ulcers of varying sizes,and partial gland loss.The TNBS group also showed increased FITC-dextran permeability in the colon,elevated mRNA expression of TNFA,IL-1b,IL-6,and IL-17,significantly upregulated MLCK,p-MLC,and PUMA protein expression,and decreased occludin protein expression.Compared with the TNBS group,the SNS group demonstrated mitigated body weight loss,longer colons,reduced FITC-dextran permeability,alleviated inflammatory cell infiltration,partial gland recovery,and reducedulcer areas.Additionally,the SNS group displayed decreased mRNA expression of TNFA,IL-1b,IL-6,and IL-17 in colonic mucosa,along with downregulated MLCK,p-MLC,and PUMA protein expression,and increased occludin protein expression.Conclusion SND reduced m

关 键 词：四逆散 溃疡性结肠炎 肠上皮屏障 肌球蛋白轻链激酶 紧密连接 

分 类 号：R285.5[医药卫生—中药学]