健脾益气方调控间隙连接蛋白43防治PD大鼠腹膜损伤的作用机制研究  

Interventional effect of spleen-strengthening and Qi-tonifying prescription on peritoneal injury in rats by regulating the gap junction protein connexin 43

作　　者：孟立锋[1,2] 杨端云 史伟 杜少鹏[1] 樊均明 MENG Lifeng;YANG Duanyun;SHI Wei;DU Shaopeng;FAN Junming(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,Guangxi,China;Combined Traditional Chinese and Western Medicine Laboratory for Controlling Organ Fibrosis,Southwest Medical University,Luzhou 646000,Sichuan,China;The First Affiliated Hospital of Chengdu Medical College,Chengdu 610500,Sichuan,China)

机构地区：[1]广西中医药大学第一附属医院,广西南宁530023 [2]西南医科大学中西医结合防治器官纤维化实验室,四川泸州646000 [3]成都医学院第一附属医院,四川成都610500

出　　处：《湖南中医杂志》2020年第7期157-161,共5页Hunan Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81560761);中国博士后基金项目(2017M612987);广西壮族自治区自然基金面上项目(2017JJA140363y)。

摘　　要：目的:研究健脾益气方通过调控间隙连接蛋白(Cx)43干预腹膜透析大鼠腹膜间皮细胞转分化的作用机制。方法:将75只大鼠随机分为5组,即空白组、模型组、中药高剂量组、中药低剂量组、西药组,除空白组大鼠外,其余各组均采用5/6肾切除联合腹腔注射4.25%腹透液复制慢性肾衰竭腹膜纤维化大鼠模型。造模成功后,中药高、低剂量组给予健脾益气方,西药组给予阿托伐他汀钙,模型组给予0.9%氯化钠注射液,空白组不做任何处理。每天灌胃1次,连续4周。干预后光镜观察各组大鼠腹膜结构变化,测量腹膜致密层厚度;免疫组化法及蛋白质印迹法检测Cx43、蛋白激酶B/哺乳动物雷帕霉素靶蛋白(AKt/m TOR)通路和腹膜间皮细胞转分化(EMT)标志蛋白表达。结果:各组大鼠腹膜厚度及Cx43、p AKt、pm TOR、α平滑肌肌动蛋白(α-SMA)、E-钙黏附蛋白(E-cadherin)阳性面积比,空白组与模型组比较,中药高、低剂量组及西药组与模型组比较,中药高剂量组与西药组、中药低剂量组比较,差异均有统计学意义(P<0.05)。各组大鼠腹膜组织Cx43、p AKt、pmTOR、α-SMA、E-cadherin蛋白表达,空白组与模型组比较,中药高剂量组与西药组比较,差异均有统计学意义(P<0.05)。结论:健脾益气方可能通过调控Cx43表达,干预Akt/m TOR信号通路介导的腹膜间皮细胞EMT进程,从而发挥保护腹膜间皮细胞、防治腹膜损伤的作用。Objective:To investigate the mechanism of action of spleen-strengthening and Qi-tonifying prescription in the intervention of peritoneal mesothelial cell transdifferentiation in a rat model of peritoneal dialysis by regulating the gap junction protein connexin 43(Cx43).Methods:A total of 75 rats were randomly divided into blank group,model group,high-and low-dose traditional Chinese medicine(TCM)groups,and Western medicine group.All rats except those in the blank group were given 5/6 nephrectomy combined with intraperitoneal injection of 4.25%peritoneal dialysis solution to establish a rat model of chronic renal failure and peritoneal fibrosis.After successful modeling,the rats in the high-and low-dose TCM groups were given spleen-strengthening and Qi-tonifying prescription,those in the Western medicine group were given atorvastatin calcium,those in the model group were given 0.9%sodium chloride injection,and those in the blank group were not given any treatment.The drugs were given by gavage once a day for 4 consecutive weeks.After intervention,a light microscope was used to observe the change in peritoneal structure and measure the thickness of peritoneal dense layer;immunohistochemistry and Western blot were used to measure the expression of Cx43,the protein kinase-B/mammalian target of rapamycin(Akt/m TOR)pathway,and marker proteins for the epithelial-to-mesenchymal transdifferentiation(EMT)of peritoneal mesothelial cells.Results:There were significant differences in peritoneal thickness and positive area ratios of Cx43,phosphorylated AKt(pAKt),phosphorylated m TOR(pmTOR),alpha-smooth muscle actin(α-SMA),and epithelial cadherin(E-cadherin)between the blank group and the model group,between the high-and low-dose TCM groups/Western medicine group and the model group,and between the high-dose TCM group and the Western medicine group or low-dose TCM group(P<0.05).There were significant differences in the protein expression of Cx43,p AKt,pm TOR,α-SMA,and E-cadherin in the peritoneal tissue between the blank grou

关 键 词：腹膜纤维化 腹膜透析 大鼠 健脾益气方 间隙连接蛋白43 实验研究 

分 类 号：R285.5[医药卫生—中药学]