激活下丘脑腹内侧核对心肌缺血再灌注损伤的影响  

Effect of activating ventromedial hypothalamus on myocardial ischemia-reperfusion injury

作　　者：李泽衍 许骁 周雨扬 周慧鑫 聂鹂庆 江洪[1] LI Ze-yan;XU Xiao;ZHOU Yu-yang;ZHOU Hui-xin;NIE Li-qing;JIANG Hong(Department of Cardiology,Renmin Hospital of Wuhan University,Cardiac Autonomic Nervous Research Center,Wuhan University,Cardiovascular Research Institute,Wuhan University,Hubei Key Laboratory of Cardiology,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院心内科,武汉大学心脏自主神经研究中心,武汉大学心血管病研究所,心血管病湖北省重点实验室,湖北省武汉市430060

出　　处：《中国心血管病研究》2021年第12期1147-1152,共6页Chinese Journal of Cardiovascular Research

基　　金：国家自然科学基金(81970287)。

摘　　要：目的探讨化学遗传激活下丘脑腹内侧核(ventromedial hypothalamus,VMH)对大鼠心肌缺血再灌注损伤后交感神经系统活性及心肌细胞的影响。方法12只Sprague-Dawley雄性大鼠按照随机数表法分为对照组(N=6)和激活组(N=6)。通过脑立体定位技术对激活组大鼠VMH注射激活病毒,对照组大鼠VMH注射空载病毒。在注射病毒3周后,两组大鼠均接受连续2周腹腔注射氯氮平一氧化氮(Clozapine-N-Oxide,CNO)。完成连续2周CNO注射后,对照组和激活组均构建心脏缺血再灌注损伤模型。免疫荧光染色脑切片观察VMH活性变化(c-fos表达情况);酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测血清去甲肾上腺素(norepinephrine,NE)浓度;苏木精-伊红染色(hematoxylin-eosin,HE)观察心肌组织改变;q RT-PCR和蛋白质印迹法(western blotting)检测心肌细胞蛋白激酶Cβ2(protein kinase Cβ2,PKCβ2)的m RNA和蛋白表达水平。结果与对照组相比,激活组VMH中c-fos表达增加[(1.00±0.11)比(1.78±0.16),P<0.01],血清NE浓度显著升高[(9.70±0.27)nmol/L比(14.46±0.17)nmol/L,P<0.01],心肌细胞排列紊乱加重、胞质嗜伊红浓染,细胞水肿显著,心肌细胞平均面积显著升高[(2769.16±169.44)μm^(2)比(7548.90±699.82)μm^(2),P<0.001],心肌PKCβ2的m RNA[(1.00±0.27)比(1.81±0.17),P<0.05]和蛋白[(1.00±0.30)比(2.50±0.24),P<0.05]表达水平显著增加。结论激活VMH可升高交感神经系统活性及恶化心肌缺血再灌注损伤。Objective To investigate the effect of activating ventromedial hypothalamus(VMH) by chemical genetic method on sympathetic nervous system activity and cardiomyocytes after myocardial ischemiareperfusion injury in rats. Methods Twelve Sprague-Dawley male rats were randomly divided into the control group(N=6) and activation group(N=6). The VMH of the activation group was injected with the activated virus by the brain stereotactic technique, and the control group was injected with the empty virus. Three weeks after the virus injection, both groups of rats received intraperitoneal injections of Clozapine-N-Oxide(CNO) for two consecutive weeks and then the cardiac ischemia-reperfusion injury models established. Rats brain sections was used to observe the alteration in VMH activity by the use of immunofluorescence stain(c-fos expression);EnzymeLinked Immunosorbent Assay(ELISA) was used to detect serum norepinephrine(NE) concentration;HematoxylinEosin staining(HE) was used to observe the alteration in myocardial tissue;q RT-PCR and Western blotting were used to detect protein kinase C β2(Protein Kinase C β2, PKC β2) m RNA and protein expression in myocardial cell. Results Compared with the control group, the expression of c-fos in VMH increased significantly [(1.78 ±0.16)VS(1.00±0.11),P<0.01] and the concentration of serum NE elevated significantly [(14.46±0.17)nmol/L vs.(9.70±0.27)nmol/L,P<0.01]. The cardiomyocytes arranged disorderly, cytoplasmic stained by eosin seriously and the cells swelled significantly. The average cardiomyocyte area increased significantly [(7548.90 ±699.82)μm^(2) vs.(2769.16±169.44)μm^(2),P<0.001];the PKC β2 m RNA expression [(1.81±0.17)vs.(1.00±0.27),P<0.05] and protein expression [(2.50±0.24)vs.(1.00±0.30),P<0.05] in cardiomyocytes increased significantly in activation group. Conclusion Activating VMH could increase sympathetic nervous system activity and worsen myocardial ischemia-reperfusion injury.

关 键 词：心脏缺血再灌注损伤 交感神经系统 下丘脑腹内侧区 蛋白激酶Cβ2 

分 类 号：R541.6[医药卫生—心血管疾病]