抵当陷胸汤对高糖诱导心肌微血管内皮细胞损伤的影响  

Effect of Didang Xianxiong Decoction on Cardiac Myocardial Microvascular Endothelial Cell Injury Induced by High Glucose

作　　者：喻锦 储全根[1,2] 李飞翔 蔡正银[1] 储俊[2] YU Jin;CHU Quan-gen;LI Fei-xiang;CAI Zheng-yin;CHU Jun(School of Traditional Chinese Medicine,Anhui University of Chinese Medicine,Hefei 230038,China;Key Laboratory of Xin'an Medicine,Ministry of Education,Anhui University of Chinese Medicine,Hefei 230038,China)

机构地区：[1]安徽中医药大学中医学院,合肥230038 [2]安徽中医药大学教育部新安医学重点实验室,合肥230038

出　　处：《中国实验方剂学杂志》2022年第9期52-58,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81774189)。

摘　　要：目的:基于高糖诱导的心肌微血管内皮细胞(CMECs)模型,观察抵当陷胸汤对其损伤的影响,并探讨其相关作用机制。方法:采用体外培养大鼠原代心肌细胞,给予33 mmol·L^(-1)葡萄糖造模,造模成功后随机分为模型组(葡萄糖终浓度为33 mmol·L^(-1))、正常组、抵当陷胸汤低剂量组(葡萄糖+5%抵当陷胸汤含药血清)、抵当陷胸汤中剂量组(葡萄糖+10%抵当陷胸汤含药血清)、抵当陷胸汤高剂量组(葡萄糖+20%抵当陷胸汤含药血清)和阿拉氯胺(ALT-711)组(葡萄糖+10%ALT-711含药血清)。采用噻唑蓝(MTT)比色法检测含药血清对CMECs增殖的影响;实时荧光定量聚合酶链式反应(Real-time PCR)检测各组c-Jun mRNA相对表达水平;蛋白免疫印迹法(Western blot)检测磷酸化蛋白质酪氨酸激酶1(p-JAK1)、磷酸化信号转导和转录激活因子1(p-STAT1)、转化生长因子-β;(TGF-β;)蛋白的表达水平。结果:与正常组比较,模型组细胞中c-Jun mRNA、p-JAK1、p-STAT1及TGF-β;蛋白的表达均显著升高(P<0.01);与模型组比较,各治疗组细胞中c-Jun mRNA表达水平显著降低(P<0.01),抵当陷胸汤中、高剂量组及ALT-711组中p-JAK1、p-STAT1蛋白表达水平均明显降低(P<0.05,P<0.01),抵当陷胸汤低剂量组变化差异无统计学意义,各用药组TGF-β;蛋白均明显降低(P<0.05,P<0.01),且抵当陷胸汤中、高剂量组较低剂量组下降明显。结论:抵当陷胸汤对高糖损伤的CMECs起到保护作用,其机制可能与降低细胞内JAK/STAT信号转导通路活性有关。Objective:This study was designed to observe the effect of Didang Xianxiong decoction on the cardiac myocardial microvascular endothelial cells(CMECs)injury,and to explore its related mechanism based on the CMECs model induced by high glucose.Method:Rat primary myocardial cells were cultured in vitro and 33 mmol·L^(-1)glucose was added for modeling.After modeling,the rats were randomly divided into model group(final glucose concentration:33 mmol·L^(-1)),normal group,Didang Xianxiong decoction low dose group(glucose+5%Didang Xianxiong decoction containing serum),Didang Xianxiong decoction medium dose group(glucose+10%Didang Xianxiong decoction containing serum),Didang Xianxiong decoction high dose group(glucose+20%Didang Xianxiong decoction containing serum)and alagebrium chloride(ALT-711)group(glucose+10%ALT-711 containing serum).The influence of drug-containing serum on the proliferation of CMECs was detected by MTT tetrazolium salt colorimetric assay.The relative mRNA expression of c-Jun was detected by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).The protein expression of phosphorylated Janus kinase 1(p-JAK1),phosphorylated signal transducer and activator of transcription 1(p-STAT1)and transforming growth factor-β;(TGF-β;)was determined by Western blot.Result:Compared with the conditions in normal group,the mRNA expression of c-Jun and protein expression of p-JAK1,p-STAT1 and TGF-β;were up-regulated in model group(P<0.01).Compared with model group,all treatment groups had decreased mRNA expression of c-Jun(P<0.01).Didang Xianxiong decoction medium and high dose groups and ALT-711 group showed reduced protein expression of p-JAK1 and p-STAT1(P<0.05,P<0.01),while there was no significant change in Didang Xianxiong decoction low dose group.TGF-β;protein expression was lowered in all treatment groups(P<0.05,P<0.01),and the decrease was more significant in Didang Xianxiong decoction medium and high dose groups than Didang Xianxiong decoction low dose group.Conclusion:Didang Xia

关 键 词：抵当陷胸汤 糖尿病心肌病 心肌微血管内皮细胞 蛋白质酪氨酸激酶1/信号转导和转录激活因子(JAK/STAT)信号通路 

分 类 号：R2-0[医药卫生—中医学] R22