熊胆粉通过抑制MLCK/pMLC2通路改善肠上皮屏障功能缓解结肠炎  

Bear bile powder alleviates colitis by improving intestinal epithelial barrier function via inhibiting MLCK/pMLC2 signaling pathway

作　　者：田瑶 梁琪 李金虎 吴雨柔 魏大能[1,2] 吴纯洁 TIAN Yao;LIANG Qi;LI Jinhu;WU Yurou;WEI Daneng;WU Chunjie(Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;Chengdu Fifth People's Hospital,Chengdu 611130,China)

机构地区：[1]成都中医药大学,成都611137 [2]成都市第五人民医院,成都611130

出　　处：《中华中医药杂志》2024年第9期4659-4664,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：四川省自然科学基金项目(No.2022NSFSC1397)。

摘　　要：目的:探究熊胆粉对结肠炎的治疗作用及机制。方法:小鼠自由饮用含3%葡聚糖硫酸钠的水溶液诱导结肠炎模型,造模成功后给予不同剂量的熊胆粉溶液(150、300、600 mg/kg)连续7 d,进行小鼠疾病活动指数(DAI)评分;测量结肠长度;HE染色观察结肠病理学变化;阿尔新蓝染色观察黏蛋白含量;FITC-葡聚糖检测肠屏障完整性;免疫荧光染色和Western Blot观察Occludin、E-cadherin、Dsg2蛋白表达;Western Blot检测MLCK/pMLC2信号通路蛋白。结果:与空白组比较,模型组小鼠体质量出现显著下降,DAI评分和组织学评分显著上升,结肠变短(P<0.01)。低、中剂量的熊胆粉可缓解体质量的下降,降低DAI评分,保护小鼠结肠组织完整性,减轻结肠的缩短,降低组织学评分(P<0.01),提高Occludin、Dsg2、E-cadherin蛋白的表达(P<0.01),减少了MLCK蛋白的表达,降低了pMLC2/MLC2蛋白的比例(P<0.01)。结论:熊胆粉通过抑制MLCK/pMLC2信号通路,保护细胞连接蛋白,修复肠屏障,而发挥抗结肠炎作用。Objective:To explore the therapeutic effect and mechanism of bear bile powder on colitis.Methods:Colitis was induced in mice by making them freely drink water solution containing 3%dextran sulfate sodium salt(DSS).After successful modeling,bear bile powder solutions of different concentrations(150,300,600 mg/kg)were given for seven consecutive days,and Disease activity index(DAI)were performed.Measurement of colon length;HE staining was used to observe the pathological changes of colon.Alcian blue staining was used to observe the number of mucins.The integrity of intestinal barrier was detected by FITC-Dextran.The expression of Occludin,E-cadherin and Dsg2 were observed by immunofluorescence staining and Western Blot.Detection of MLCK/pMLC2 signaling pathway protein by Western Blot.Results:Compared with the control group,mice in the model group showed a significant decrease in body weight,a rise in DAI score and histological score,and a shortened colon(P<0.01).Low and medium doses of bear bile powder alleviated the decrease in body weight,reduced DAI scores,protected the integrity of mice colon tissues,alleviated the shortening of the colon,and reduced histological scores(P<0.01),increased the expression of the proteins Occludin,Dsg2,and E-cadherin(P<0.01),and reduced the expression of the MLCK protein,and reduced the pMLC2/MLC2 protein ratio(P<0.01).Conclusion:Bear bile powder plays an anti-colitis role by inhibiting MLCK/pMLC2 signaling pathway and protecting cell connexin repairing intestinal barrier.

关 键 词：熊胆粉 结肠炎 黏蛋白 肠道屏障 MLCK/pMLC2通路 

分 类 号：R285.5[医药卫生—中药学]