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
田瑶 1梁琪 1李金虎 1吴雨柔 1魏大能 2吴纯洁1
作者信息
- 1. 成都中医药大学,成都 611137
- 2. 成都中医药大学,成都 611137;成都市第五人民医院,成都 611130
- 折叠
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.
Abstract
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Key words
Bear bile powder/Colitis/Mucin/Intestinal barrier/MLCK/pMLC2 pathway引用本文复制引用
基金项目
四川省自然科学基金项目(2022NSFSC1397)
出版年
2024
国家科技期刊平台
NSTL
万方数据