经皮耳缘迷走神经刺激对大鼠射血分数保留性心力衰竭模型心肌结构重塑和电重塑及凋亡的干预作用  

Intervention effect of transcutaneous auricular vagus nerve stimulation on myocardial structural remodeling, electrical remodeling and apoptosis in rat heart failure model with preserved ejection fraction

作　　者：彭明 李玉凯 王岚[1] 黄粮[1] 成忠[1] 肖杰 Peng Ming;Li Yukai;Wang Lan;Huang Liang;Cheng Zhong;Xiao Jie(Department of Cardiovascular Medicine,Wuhan Fourth Hospital,Wuhan 430033,China)

机构地区：[1]武汉市第四医院心血管内科,武汉430033

出　　处：《中华老年医学杂志》2023年第3期334-340,共7页Chinese Journal of Geriatrics

基　　金：湖北省卫生健康委员会科研基金资助项目(WJ2019M041)。

摘　　要：目的 观察经皮耳缘迷走神经刺激对大鼠射血分数保留性心力衰竭模型心肌结构重塑、电重塑及凋亡的干预作用,并探讨这种作用与氧化应激反应的关系。方法 通过对SD大鼠建立动静脉瘘后2周使用结扎的方法封闭动静脉瘘,在短期内增加心脏容量负荷从而构建大鼠射血分数保留性心力衰竭模型。40只大鼠随机分为4组,每组各10只:假手术组、腹主动脉-下腔静脉瘘+封闭组(AVF+L)、腹主动脉-下腔静脉瘘+封闭+经皮耳缘迷走神经刺激组(AVF+L+tVNS)及腹主动脉-下腔静脉瘘+封闭+经皮耳缘迷走神经刺激+乙酰胆碱M_(2)受体拮抗剂使用组(AVF+L+tVNS+M-)。AVF+L+tVNs组在AVF+L基础上进行经皮耳缘迷走神经刺激。AVF+L+tVNS+M-组在AVF+L+tVNs组基础上每天尾静脉注射乙酰胆碱M_(2)受体拮抗剂美索曲明(0.5 mg/kg)。通过心脏超声仪和心脏电生理刺激仪获得各组大鼠心脏结构重塑及电重塑的相关参数。酶联免疫吸附(ELISA)法检测各组大鼠B型尿钠肽前体(NT-proBNP)及氧化应激相关指标的值。苏木素伊红(HE)染色观察心肌结构破坏、细胞排列紊乱及炎性细胞浸润情况。缺口末端标记法(TUNL)、反转录聚合酶链反应(RT-PCR)和免疫印迹(Western blotting)法检测心肌B细胞淋巴瘤/白血病基因-2(BCL-2)、BCL-2基因家族中细胞凋亡促进基因(BAX)mRNA及蛋白的表达。结果 AVF+L组大鼠出现以心室壁肥厚及舒张功能不全为特征性的心力衰竭,且左心室射血分数(LVEF)均>50%,考虑大鼠射血分数保留性心力衰竭模型构建成功。HE染色提示AVF+L组大鼠心肌组织结构破坏、细胞排列紊乱及炎性细胞浸润明显,而AVF+L+tVNs组大鼠较AVF+L组心肌组织病理改变均明显缓解。与AVF+L组比较,AVF+L+tVNs组大鼠NT-proBNP值降低[(301.25±16.07)ng/L比(79.33±5.63)ng/L,P<0.05]、E/A值升高(1.28±0.06比1.66±0.05,P<0.05)、BCL-2 mRNA的表达升高[0.08(0.07,0.08)比0.70(0.64,0.76),P<0.05]及BCL-Objective To observe the effect of percutaneous auricular vagus nerve stimulation on myocardial structural remodeling,electrical remodeling and apoptosis in rats of heart failure with preserved ejection fraction,and to explore the relationship between this effect and oxidative stress.Methods The arteriovenous fistula was closed by ligation two weeks after establishment in SD rat.By increasing cardiac volume load in the short term,a rat model of heart failure with preserved ejection fraction was constructed.Forty rats were randomly divided into four groups,with 10 rats in each group:sham operation group(S),abdominal aorta-inferior vena cava fistula+closure group(AVF+L),abdominal aorta-inferior vena cava fistula+closure+percutaneous auricular vagus nerve stimulation group(AVF+L+tVNS)and abdominal aorta-inferior vena cava fistula+closure+percutaneous auricular vagus nerve stimulation+acetylcholine M_(2) receptor antagonist group(AVF+L+tVNS+M-).Rats in the AVF+L+tVNS group received percutaneous vagal nerve stimulation on the basis of those in the AVF+L group.Rats in the AVF+L+tVNS+M-group received daily injection of acetylcholine M_(2) receptor antagonist mesotramine(0.5mg/Kg)into tail vein on the basis of those in the AVF+L+tVNS group.The parameters of cardiac structural remodeling and electrical remodeling in each group were obtained by cardiac ultrasound and cardiac electrophysiological stimulator.Enzyme-linked immunosorbent assay(ELISA)was used to detect the values of B-type natriuretic peptide precursor(NT-proBNP)and oxidative stress-related indicators in each group.hematoxylin-eosin(HE)staining was used to observe the damage of myocardial structure,disorder of cell arrangement and infiltration of inflammatory cells.Cardiomyocyte apoptosis was observed by TdT-mediated dUTP nick end labeling(TUNEL)staining and apoptosis index was calculated.reverse transcription-polymerase chain reaction(RT-PCR)and Western blotting were used to detect the mRNA and protein expression of B cell lymphoma/leukemia-2(BCL-2)and apoptos

关 键 词：迷走神经 每搏输出量 心力衰竭 心里重构 细胞凋亡 氧化性应激 

分 类 号：R541.6[医药卫生—心血管疾病] R-332[医药卫生—内科学]