线粒体ATP敏感性钾通道在降钙素基因相关肽减轻新生大鼠心肌细胞缺氧复氧损伤中的作用  

Role of mitochondrial K ATP channel in CGRP-induced reduction of cardiomyocyte anoxia-reoxygenation injury in rats

作　　者：王鑫[1] 张文颉[1] 田首元[1] WANG Xin;ZHANG Wenjie;TIAN Shouyuan(Department of Anesthesiology,the First Hospital of Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]山西医科大学第一医院麻醉科,太原市030001

出　　处：《临床麻醉学杂志》2020年第1期72-76,共5页Journal of Clinical Anesthesiology

基　　金：山西医科大学第一医院青年基金(YQ161705)。

摘　　要：目的评价线粒体ATP敏感性钾通道(mitoK ATP通道)在降钙素基因相关肽(CGRP)减轻新生大鼠心肌细胞缺氧复氧损伤中的作用。方法取原代培养的新生1~3 d SD大鼠心肌细胞,在含有10%胎牛血清培养基中培养,细胞接种于6孔细胞培养板,采用随机数字表法分为5组(n=10):正常对照组(C组)、缺氧复氧组(AR组)、CGRP+缺氧复氧组(CGRP组)、CGRP+mitoK ATP通道阻滞剂(5-HD)+缺氧复氧组(5-HD组)、CGRP+CGRP受体阻滞剂CGRP 8-37+缺氧复氧组(CGRP 8-37组)。除C组,其余4组均缺氧3 h,复氧2 h;CGRP组缺氧前2 h加入终浓度为0.1 nmol/L CGRP;5-HD组于缺氧前2.5 h加入终浓度为500μmol/L 5-HD,30 min后加入终浓度为0.1 nmol/L CGRP;CGRP 8-37组于缺氧前2.5 h加入终浓度为1 nmol/L CGRP 8-37,30 min后加入终浓度为0.1 nmol/L CGRP。于缺氧复氧后检测心肌细胞凋亡情况,计算凋亡指数(AI)、乳酸脱氢酶(LDH)活性,JC-1荧光法测定线粒体膜电位(Δψm)的变化,即JC-1多聚体/单聚体的比值。结果AR组AI明显高于C组(P<0.01)LDH活性明显强于C组(P<0.01),JC-1多聚体/单聚体的比值明显低于C组(P<0.01)。CGRP组和5-HD组AI明显低于AR组(P<0.05),LDH活性明显弱于AR组(P<0.01),JC-1多聚体/单聚体的比值明显高于AR组(P<0.01)。5-HD组和CGRP 8-37组AI明显高于CGRP组(P<0.01),LDH活性明显强于CGRP组(P<0.01),JC-1多聚体/单聚体的比值明显低于CGRP组(P<0.01)。结论CGRP可以激活新生大鼠心肌细胞膜CGRP受体,通过激活mitoK ATP通道,减少心肌细胞缺氧复氧损伤。Objective To evaluate the role of mitochondrial K ATP channel(mitoK ATP)in CGRP-induced reduction of cardiomyocyte anoxia-reoxygenation injury in rats.Methods Cardiomyocytes were obtained from 1-3 day old Sprague-Dawley rats and cultured in the culture medium containing 10%bovine calf serum and then seeded onto 6-well plates at a density of 10×105/ml(3 ml/well).The cells were randomly divided into 5 groups(n=10 each):control group(group C),anoxia-reoxygenation group(group AR);CGRP+AR group(group CGRP);CGRP+mitoK ATP channel blocker(5-HD)+AR group(group 5-HD);CGRP+CGRP receptor blocker(CGRP 8-37)+AR group(group CGRP 8-37).Except group C,the other groups were anoxia for 3 hours and reoxygenated for 2 hours.In group CGRP,CGRP(final concentration 0.1 nmol/L)was added to the culture medium before 2 h anoxia.In group 5-HD,5-HD(final concentration 500μmol/L)was added to the culture medium 2.5 h before anoxia,and CGRP(final concentration 0.1 nmol/L)was added after 30 min.In group CGRP 8-37,CGRP 8-37(final concentration 1 nmol/L)was added to the culture medium 2.5 h before anoxia,and CGRP(final concentration 0.1 nmol/L)was added after 30 min.At the end of reoxygenation,the level of lactate dehydrogenase(LDH)in the supernatant was detected,the changes in mitochondrial membrane potential were assessed by JC-1 fluorescence assay,the cell apoptosis was detected by Tunnel.Results Compared with group C,the amount of LDH released and AI were significantly increased,JC-1 monomer/aggregate ratio were significantly decreased in group AR(P<0.01).Compared with group AR,the amount of LDH released and AI were significantly decreased,JC-1 monomer/aggregate ratio were significantly increased in group CGRP(P<0.01).Compared with group CGRP,the amount of LDH released and AI were significantly increased,JC-1 monomer/aggregate ratio were significantly decreased in group 5-HD and CGRP 8-37(P<0.01).Conclusion CGRP can reduce A/R-induced injury to neonatal rat cardiomyocytes through activating mitoK ATP channel via combing with CGRP receptor.

关 键 词：降钙素基因相关肽 细胞低氧 肌细胞 心脏 线粒体膜 离子通道 钾 

分 类 号：R-332[医药卫生] R54