降钙素基因相关肽对高糖下新生大鼠心肌细胞缺氧复氧损伤的影响  

Effect of calcitonin gene-related peptide on anoxia-reoxygenation induced injury to neonatal rat cardiomyocytes incubated in high glucose medium

作　　者：陈璐[1] 赵鑫[1] 郭政[1] 

机构地区：[1]山西医科大学第二医院麻醉科,太原市030001

出　　处：《中华麻醉学杂志》2014年第10期1185-1188,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（30972860）

摘　　要：目的 评价降钙素基因相关肽（CGRP）对高糖下新生大鼠心肌细胞缺氧复氧（A/R）损伤的影响.方法 取原代培养的出生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,接种密度10×10^5/ml,3 ml/孔,采用随机数字表法,将其分为5组（n=9）,正常糖浓度对照组（NG组）用正常糖浓度（葡萄糖5.5 mmol/L）培养基孵育细胞72 h;高糖对照组（HG组）用高糖（葡萄糖25.0 mmol/L）培养基孵育细胞72 h;HG＋ A/R组用高糖培养基孵育细胞72 h后,行缺氧3h,复氧2h;HG＋ A/R＋ CGRP组用高糖培养基孵育细胞72 h后加入CGRP（终浓度为10^-8 mol/L）,1h后行缺氧3h,复氧2h;HG＋ MR＋ CGRP＋ CGRP8-37组用高糖培养基孵育细胞72 h后加入CGRP（终浓度为10^-8 mol/L）和CGRP8-37（CGRP受体拮抗剂,终浓度10^-7 mol/L）,1h后行缺氧3h,复氧2h.处理后收集细胞,采用TUNEL法检测心肌细胞凋亡情况,计算凋亡指数（AI）.于缺氧3h和复氧2h时分别收集细胞培养液,测定乳酸脱氢酶（LDH）活性.结果 与NG组比较,HG组AI、细胞培养液LDH活性升高（P＜0.01）;与HG组比较,HG＋ A/R组AI、细胞培养液LDH活性升高（P＜0.01）;与HG＋ A/R组比较,HG＋ A/R＋ CGRP组AI、细胞培养液LDH活性降低（P＜0.01）,HG＋ A/R＋ CGRP＋CGRP8-37组上述指标比较差异无统计学意义（P＞0.05）;与HG＋ A/R＋ CGRP组比较,HG＋ A/R＋ CGRP＋ CGRP8-37组AI、细胞培养液LDH活性升高（P＜0.01）.结论 CGRP可通过与CGRP受体结合减轻高糖下新生大鼠心肌细胞缺氧复氧损伤.Objective To evaluate the effect of calcitonin gene-related peptide （CGRP） on anoxia- reoxygenation （A/R）-induced injury to neonatal rat cardiomyocytes incubated in high glucose medium. Methods Cardiomyocytes were obtained from 1-3-day old Sprague-Dawley rats and cultured in the culture medium containing 15% bovine calf serum and then seeded onto 6-well plates at a density of 10 × 10^5/ml （3 ml/well）. The cells were randomly divided into 5 groups （ n = 9 each） ： normal glucose medium control group （NG group）, high glucose medium group （HG group）, high glucose medium ＋ A/R group （HG＋ A/R group）, high glucose medium ＋ A/R ＋ CGRP group （HG ＋ A/R ＋ CGRP group）, and high glucose medium ＋ A/R ＋ CGRP ＋ CGRP8-37 group （HG ＋ A/R ＋ CGRP ＋ CGRP8-37 group）. The cells were incubated in .normal glucose （5.5 mmol/L） medium for 72 h in NG group. In HG group, the ceils were incubated in high glucose （25.0 mmol/L） medium for 72 h. In HG ＋ A/ R group, the cells were incubated in high glucose medium tbr 72 h and then exposed to 3 h of anoxia followed by 2 h of reoxygenation. In group HG ＋ A/R ＋ CGRP, the cells were incubated in high glucose medium for 72 h, CGRP （final concentration 10-s mol/L） was then added to the culture media and the cells were incubated for 1 h and then underwent A/R. In HG ＋ A/R ＋ CGRP ＋ CGRP8-37 group, the cells were incubated in high glucose medium for 72 h, CGRP （final concentration 10^-8 mol/L） was then added to the culture media and the cells were incubated for 1 h and then underwent A/R. In HG ＋ A/R ＋ CGRP ＋ CGRP8-37 group, the cells were incubated in high glucose medium for 72 h, CGRP8-37 （final concentration 10-8 tool/L） and CGRP8-37 （CGRP receptor antagonist, final concentration 10^-7 mol/L） was then added to the culture nmdia and the cells were incubated for 1 h and then underwent A/R. Apoptosis in cardiomyocytes was detected using TUNEL and apoptosis index （AI） was calculated. The lactate deh

关 键 词：降钙素基因相关肽 糖尿病 细胞低氧 肌细胞 心脏 婴儿 新生 

分 类 号：R722.1[医药卫生—儿科]