补中益气方通过MLCK/p-MLC_(20)通路调节慢性腹泻大鼠结肠动力的机制研究  

Mechanism of Buzhong Yiqi Prescription(补中益气方)on Inhibiting Colonic Smooth Muscle Contraction through Downregulation of MLCK/p-MLC_(20)Pathway in Chronic Diarrhea Rats

作　　者：贾梦迪 张声生[1] 赵鲁卿[1] 卢小芳[1] 朱泠霏 JIA Mengdi;ZHANG Shengsheng;ZHAO Luqing;LU Xiaofang;ZHU Lingfei(Digestive Disease Center,Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University,Beijing 100010,China)

机构地区：[1]首都医科大学附属北京中医医院消化中心,北京100010

出　　处：《辽宁中医杂志》2024年第10期180-186,共7页Liaoning Journal of Traditional Chinese Medicine

基　　金：国家自然科学青年基金项目(82104745)。

摘　　要：目的探讨补中益气方对慢性腹泻大鼠MLCK/p-MLC_(20)通路介导的结肠动力的影响。方法大鼠随机分为正常组,模型组和补中益气低、中、高剂量组。模型组及补中益气方组应用大黄灌胃的方法建立慢性腹泻大鼠模型。慢性腹泻模型建立成功后,补中益气汤颗粒低剂量:0.4725 g/(kg·d),中剂量:0.945 g/(kg·d),高剂量:1.89 g/(kg·d),正常组和模型组则每天给予等体积的生理盐水灌胃。在治疗结束时,检测各组大鼠一般情况如体质量、饮食量、悬空拉尾抵抗实验、免疫指标(胸腺指数、脾脏指数)变化和24小时粪便含水量。进一步探讨机制,采用离体张力测定灌流系统,测定各组大鼠结肠纵行平滑肌肌条收缩力(colonic longitudinal smooth muscle strips,CLSMs)和结肠环型平滑肌肌条(colonic circular smooth muscle strips,CCSMs)的变化,并蛋白印迹法测定磷酸化肌球蛋白轻链(phosphorylation level of myosin light chain,p-MLC_(20))水平及实时荧光定量PCR法检测大鼠结肠组织肌球蛋白轻链激酶(myosin light chain kinase,MLCK)的表达。结果各组大鼠一般情况比较有统计学意义(P<0.05),与正常组比较,模型组一般情况表现为(体质量下降:P<0.01;饮食量减少:P<0.01;悬空拉尾抵抗实验时间缩短:P<0.01;胸腺指数降低:P<0.01;脾脏指数降低:P<0.01),与模型组比较,补中益气低、中、高剂量组一般情况明显改善(体质量增加:低剂量:P<0.05,中、高剂量:P<0.01;饮食量增多:低剂量:P<0.05,中、高剂量:P<0.01;悬空拉尾抵抗实验时间延长:低剂量:P<0.05,中、高剂量:P<0.01;胸腺指数升高:P<0.01;脾脏指数升高:P<0.01)。各组大鼠粪便含水量差异有统计学意义(P<0.01),与正常组比较,模型组粪便含水量明显增加(P<0.01),与模型组比较,补中益气中、高剂量组粪便含水量明显下降(P<0.01)。同时,正常组、模型组和补中益气中剂量组大鼠结肠CLSMs和CCSMs收缩差异有统计学意义(P<0.05),�Objective This study was designed to explore the effect of Buzhong Yiqi Prescription(补中益气方,BZYQ)in chronic diarrhea rats as well as the potential mechanisms of MLCK/p-MLC_(20)related pathway.Methods Wistar rats were randomly divided into five groups:the normal group,the model group and the BZYQ low,middle and high dose groups.The rats in the model and BZYQ low,middle and high dose groups were administered with Dahuang(rhubarb)induce chronic diarrhea.The BZYQ low,middle and high dose groups were then given BZYQ[0.4725 g/(kg·d),0.945 g/(kg·d),1.89 g/(kg·d)]for 7 days,respectively.The control and model groups received the same volume of distilled water.After treatment,the body weights,food intake,tail suspension resistance test and immunity index(thymus index,spleen index)response of common symptoms were observed and fecal water content was detected.Meanwhile,the contraction of isolated colonic longitudinal smooth muscle strips(CLSMs)and colonic circular smooth muscle strips(CCSMs)were observed and phosphorylation level of myosin light chain(p-MLC20)was determined by western blot and myosin light chain kinase(MLCK)by Real time fluorescence quantitative PCR method.Results The general situation of rats in each group was statistically significant(P<0.05).Compared with those in the normal group,the common symptoms in the model group were clearly manifested(body weight loss:P<0.01;reduced food intake:P<0.01;shortened time of experimental period of tail suspension resistance test:P<0.01;decreased thymus index:P<0.01,decreased spleen index:P<0.01).Compared with those of the model group,the common symptoms obviously improved in BZYQ low,middle and high dose groups[body weight loss:low dose group(P<0.05);middle and high dose groups(P<0.01);increased food intake:low dose(P<0.05);middle and high dose(P<0.01);lengthened time of experimental period of tail suspension resistance test:low dose(P<0.05);middle and high groups(P<0.01);increased thymus index:P<0.01,increased spleen index:P<0.01].There was difference in fe

关 键 词：慢性腹泻 补中益气方 动力 肌球蛋白轻链 肌球蛋白轻链激酶 

分 类 号：R285.5[医药卫生—中药学]