|
| 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 KATP channel in CGRP-induced reduction of cardiomyocyte anoxia-reoxygenation injury in rats |
| |
| DOI：10.12089/jca.2020.01.017 |
| 中文关键词: 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 |
| 英文关键词: Calcitonin gene-related peptide Cell hypoxia Myocytes Cardiac Mitochondrial membranes Ion Channels Potassium |
| 基金项目:山西医科大学第一医院青年基金(YQ161705) |
|
| 摘要点击次数: 2786 |
| 全文下载次数: 656 |
| 中文摘要: |
目的 评价线粒体ATP敏感性钾通道(mitoKATP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+mitoKATP通道阻滞剂(5-HD)+缺氧复氧组(5-HD组)、CGRP+CGRP受体阻滞剂CGRP8-37+缺氧复氧组(CGRP8-37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500 μmol/L 5-HD,30 min后加入终浓度为0.1 nmol/L CGRP;CGRP8-37组于缺氧前2.5 h加入终浓度为1 nmol/L CGRP8-37,30 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CGRP8-37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mitoKATP通道,减少心肌细胞缺氧复氧损伤。 |
| 英文摘要: |
Objective To evaluate the role of mitochondrial KATP channel (mitoKATP) 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+mitoKATP channel blocker(5-HD)+AR group (group 5-HD); CGRP+CGRP receptor blocker (CGRP8-37)+AR group (group CGRP8-37). 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 500 μmol/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 CGRP8-37, CGRP8-37(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 CGRP8-37 (P<0.01).
Conclusion CGRP can reduce A/R-induced injury to neonatal rat cardiomyocytes through activating mitoKATP channel via combing with CGRP receptor. |
|
查看全文
查看/发表评论 下载PDF阅读器 |
| 关闭 |
|
|
|
