肾衰Ⅱ号方对慢性肾功能衰竭模型大鼠肠道紧密连接蛋白的影响  

Effect of Shenshuai FormulaⅡ(肾衰Ⅱ号方) on Intestinal Tight Junction Protein in Chronic Renal FailureModel Rats

作　　者：姚东升[1] 徐琳[1,2] 王骆冰[1] 邵命海[1] 叶朝阳 王琛[1,2] YAO Dongsheng;XU Lin;WANG Luobing;SHAO Minghai;YE Chaoyang;WANG Chen(Shuguang Hospital Affiated to ShanghaiUniversity of Traditional Chinese Medicine,Shanghai,201203;Institute of Chinese Medicine and Nephropathy,Shanghai University of Traditional Chinese Medlicine)

机构地区：[1]上海中医药大学附属曙光医院,上海市201203 [2]上海中医药大学中医肾病研究所

出　　处：《中医杂志》2021年第10期898-903,共6页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(81704015);上海市浦东新区卫生系统重点学科群建设项目(PWZxq2017-07);上海市进一步加快中医药事业发展三年行动计划[ZY(2018-2020)-CCCX-2003-08,ZY(2018-2020)-FWTX-7005]。

摘　　要：目的探讨肾衰Ⅱ号方治疗慢性肾功能衰竭的可能作用机制。方法SD大鼠采用5/6肾切除方法制备慢性肾功能衰竭大鼠模型,将造模成功的30只大鼠随机分为模型组、肾衰Ⅱ号方组及双歧三联活菌组各10只,另设空白组大鼠10只。肾衰Ⅱ号方组给予浓度为6.09g/ml肾衰Ⅱ号方药液10ml/kg灌胃,双歧三联活菌组大鼠给予双歧三联活菌17.86mg/(kg·d)灌胃,空白组、模型组大鼠给予2ml生理盐水灌胃,均每日1次,连续60天。取材前实验大鼠禁食5h后灌服内毒素(LPS)10mg/kg,3.5h后大鼠腹腔麻醉取血分离血清,10%甲醛固定小肠。检测血肌酐、尿素氮及LPS含量,肠组织采用HE染色观察病理结构,检测肠组织闭锁蛋白(Occludin)、闭锁小带蛋白1(ZO-1)、肌球蛋白轻链激酶(MLCK)、肌球蛋白轻链(MLC)蛋白及其mRNA表达,磷酸化肌球蛋白轻链(p-MLC)蛋白表达。结果与空白组比较,模型组大鼠血肌酐、尿素氮、LPS水平升高,MLCK、p-MLC蛋白及MLCK mRNA表达亦升高,ZO-1、Occludin蛋白及mRNA表达降低(P<0.05),病理显示小肠绒毛变短变宽,局部隐窝拉长,腺体增殖不活跃。与模型组比较,肾衰Ⅱ号方组血肌酐、尿素氮、LPS水平降低,MLCK蛋白及mRNA、p-MLC蛋白表达降低,ZO-1、Occludin蛋白升高(P<0.05),肠道病理变化减轻。结论肾衰Ⅱ号方可能通过下调肠组织MLCK的转录与表达,降低MLC磷酸化水平修复紧密连接结构,从而调整肠道屏障功能减少肠源性内毒素入血,达到治疗慢性肾功能衰竭的目的。Objective To explore the possible mechanism of Shenshuai FormulaⅡ(肾衰Ⅱ号)in treating chronic kidney failure(CKF).Methods CKF rats model was established by 5/6 nephrectomy in SD rats.Thirty successfully modeled rats were randomly divided into a model group,a Shenshuai FormulaⅡgroup,and bifid triple viable group,with 10 rats in each group;a blank group of 10 rats was also set.Concentration of 6.09 g/ml Shenshuai FormulaⅡat dose of 10 ml/kg and concentration of 17.86 mg/(kg·d)bifid triple viable were given by gavage to the rats in the corresponding groups;2 ml normal saline was given by gavage to the rats in the blank group and model group;all interventions were performed once daily for 60 days.The rats were fasted for five hours before being fed with 10 mg/kg endotoxin(LPS);after 3.5 hours,the rats were anesthetized in the abdominal cavity to collect and separate serum,and the small intestine was fixed with 10%formalin.The serum creatinine,urea nitrogen and LPS content were assessed.HE staining method was used to observe the pathological structure of intestinal tissue.The intestinal tissue atresia protein(Occludin),zonula atresia protein 1(ZO-1),myosin light chain kinase(MLCK),muscle globulin light chain(MLC)protein and the mRNA expression,and phosphorylated myosin light chain(p-MLC)protein expression were detected.Results Compared to the blank group,the model group had higher levels of serum creatinine,urea nitrogen and LPS content,higher expression of MLCK,p-MLC protein and MLCK mRNA,and lower expression of ZO-1 and Occludin protein and mRNA(P<0.05);and the pathological results showed that small intestinal villi became shorter and wider;local crypts elongated;and gland proliferation was not active.Compared to the model group,Shenshuai FormulaⅡgroup had lower serum creatinine,urea nitrogen and LPS content,lower expression of MLCK protein and mRNA and p-MLC protein,and higher expression of ZO-1 and Occludin protein(P<0.05);and the intestinal pathological results were improved.Conclusion Shenshuai For

关 键 词：慢性肾功能衰竭 肾衰Ⅱ号方 紧密连接蛋白 肠道屏障 内毒素 

分 类 号：R285.5[医药卫生—中药学]