真武汤经I_(to)/Kv通路对心力衰竭心肌细胞电重构的影响  

Effect of Zhenwu Decoction on electrical remodeling of cardiomyocytes in heart failure via I_(to)/Kv channels

作　　者：车驰 王晓琳 陈智勇 郑美群 唐伟 鲁宗琼 郭佳帅 黄婉清 田鑫 李林[1,2] CHE Chi;WANG Xiao-lin;CHEN Zhi-yong;ZHENG Mei-qun;TANG Wei;LU Zong-qiong;GUO Jia-shuai;HUANG Wan-qing;TIAN Xin;LI Lin(Jiangxi University of Chinese Medicine,Nanchang 330006,China;the Affiliated Hospital of Jiangxi University of Chinese Medicine,Nanchang 330006,China)

机构地区：[1]江西中医药大学,江西南昌330006 [2]江西中医药大学附属医院,江西南昌330006

出　　处：《中国中药杂志》2023年第13期3565-3575,共11页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81960820);江西省自然科学基金项目(20202BABL206141)。

摘　　要：探讨真武汤通过瞬时外向钾电流(I_(to))/电压门控钾(Kv)通路来调节电重构以治疗心力衰竭的可能作用机制。取5只正常SD大鼠灌服真武汤颗粒剂制备含药血清,另取7只正常SD大鼠灌服等量蒸馏水制备空白血清,常规传代H9c2心肌细胞,血管紧张素Ⅱ(AngⅡ)作用24 h后,分别采取2%、4%、8%含药血清,辛伐他汀(simvastatin,SIM)及氯化钡(BaCl_(2))干预细胞24 h,设为对照组[N,10%空白血清+90%高糖培养基(DMEM-H)];模型组(M,AngⅡ+10%空白血清+90%DMEM-H);真武汤含药血清低剂量组(Z1,AngⅡ+真武汤2%含药血清+8%空白血清+90%DMEM-H);真武汤含药血清中剂量组(Z2,AngⅡ+真武汤4%含药血清组+6%空白血清+90%DMEM-H);真武汤含药血清高剂量组(Z3,AngⅡ+真武汤8%含药血清组+2%空白血清+90%DMEM-H);诱导剂组(YD,AngⅡ+SIM+10%空白血清+90%DMEM-H);抑制剂组(YZ,AngⅡ+BaCl_(2)+10%空白血清+90%DMEM-H)。ELISA检测各组细胞提取液中ANP水平,实时荧光定量PCR技术检测ANP、Kv1.4、Kv4.2、Kv4.3、DPP6、KChIP2的mRNA相对表达水平,采用Western blot法检测Kv1.4、Kv4.2、Kv4.3、DPP6、KChIP2蛋白表达,全细胞膜片钳技术检测I_(to)。结果显示,低、中、高剂量真武汤均能有效降低心肌细胞表面积。与M组比较,Z1、Z2、Z3、YD组ANP含量及mRNA水平显著下降,Kv4.2、Kv4.3、DPP6、KChIP2蛋白表达及mRNA水平升高,Kv1.4蛋白表达及mRNA水平降低,以真武汤高剂量组效果最显著;与N组比较,真武汤组I_(to)峰值电流、电流密度明显上升。表明真武汤可以通过改善Kv1.4、Kv4.2、Kv4.3、KChIP2、DPP6蛋白表达趋势及诱导I_(to)调控电压门控钾通道,调节心肌结构重构和电重构,这可能是其治疗心力衰竭及相关心律失常的机制之一。This study aimed to investigate the underlying mechanism of Zhenwu Decoction in the treatment of heart failure by regulating electrical remodeling through the transient outward potassium current(I_(to))/voltage-gated potassium(Kv) channels.Five normal SD rats were intragastrically administered with Zhenwu Decoction granules to prepare drug-containing serum,and another seven normal SD rats received an equal amount of distilled water to prepare blank serum.H9c2 cardiomyocytes underwent conventional passage and were treated with angiotensin Ⅱ(AngⅡ) for 24 h.Subsequently,2%,4%,and 8% drug-containing serum,simvastatin(SIM),and BaCl_(2) were used to interfere in H9c2 cardiomyocytes for 24 h.The cells were divided into a control group [N,10% blank serum + 90% high-glucose DMEM(DMEM-H)],a model group(M,AngⅡ + 10% blank serum + 90% DMEM-H),a low-dose Zhenwu Decoction-containing serum group(Z1,AngⅡ + 2% drug-containing serum of Zhenwu Decoction + 8% blank serum + 90% DMEM-H),a medium-dose Zhenwu Decoction-containing serum group(Z2,AngⅡ + 4% drug-containing serum of Zhenwu Decoc-tion + 6% blank serum + 90% DMEM-H),a high-dose Zhenwu Decoction-containing serum group(Z3,AngⅡ + 8% drug-containing serum of Zhenwu Decoction + 2% blank serum + 90% DMEM-H),an inducer group(YD,AngⅡ + SIM + 10% blank serum + 90% DMEM-H),and an inhibitor group(YZ,AngⅡ + BaCl_(2) + 10% blank serum + 90% DMEM-H).The content of ANP in cell extracts of each group was detected by ELISA.The relative mRNA expression levels of ANP,Kv1.4,Kv4.2,Kv4.3,DPP6,and KChIP2 were detected by real-time quantitative PCR.The protein expression of Kv1.4,Kv4.2,Kv4.3,DPP6,and KChIP2 was detected by Western blot.I_(to) was detected by the whole cell patch-clamp technique.The results showed that Zhenwu Decoction at low,medium,and high doses could effectively reduce the surface area of cardiomyocytes.Compared with the M group,the Z1,Z2,Z3,and YD groups showed decreased ANP content and mRNA level,increased protein and mRNA expression of Kv4.2,Kv4.3,DPP6,and KChIP

关 键 词：心力衰竭 真武汤 电压门控钾通路 电重构 瞬时外向钾电流 

分 类 号：R285.5[医药卫生—中药学]