极化液、美托洛尔对缺氧-再给氧心肌保护作用的实验研究

STUDY OF THE PROTECT EFFECT OF GLUCOSE-INSULIN-POTASSIUM AND METOPROLOL ON CULTURED NEONATAL RAT ANOXIA-REOXYGENATION MYOCARDIAL CELL

目的探讨极化液、美托洛尔对缺氧-再给氧心肌细胞的保护作用。方法用ACAS570粘附式细胞仪,以荧光探针技术(Lndo-1-AM)激光共聚焦扫描及免疫组化、灰度分析观察极化液、美托洛尔对培养心肌细胞缺氧-再给氧(A/R)时细胞内钙(Ca2+)荧光强度、乳酸脱氢酶(LDH)变化、超氧化物岐化酶(SOD)活性、丙二醛(MDA)含量及c-Fos表达的影响。结果心肌细胞A/R后,细胞内Ca2+荧光强度升高,对照组荧光强度和荧光比率分别为:167.3±32.8,0.3±0.04,A/R组为:562.3±128.6,97.6±12.8,p<0.01。细胞培养液中LDH、MDA含量增高,SOD活性、平均灰度值降低,p<0.01。极化液组、美托洛尔组和二药合用组,上述改变得到明显改善,以二药合用组最显著,Ca2+荧光强度显著降低为275±53.6,2.7±1.3,细胞培养液中LDH、MDA含量降低,SOD活性、平均灰度值显著增高,p<0.01。结论极化液、美托洛尔有阻止Ca2+内流,减轻Ca2+超载及明显抑制培养心肌细胞A/R的c-Fos表达、降低心肌A/R的损伤的作用。

Objective To study the protect effect of Glucose-Insulin-Potassium（GIK）and metoprolol on cultured neonatal rat anoxia-reoxygenation cardiac myocytes.Methods Cultured myocytes of neonatal rats have been divited into five groups：control group ;anoxia-reoxygenation（A/R） group;Treatment groups（Glucose-Insulin-Potassium group;metoprolol group;concomitant group）.The concentration of lactate dehydrogenaseand（LDH）,the actives of SOD,the contents of cellular melondadehyde（MDA）and the expression of c-fos protein was evaluated.The free calcium（Ca2＋） level was measured through adherent cell analysis and sorting cytometer.Results Cellular Ca2＋ raising speed was fast after A/R,Control group：167.3±32.8,0.3±0.04,A/R group：562.3±128.6,97.6±12.8,p0.01;The actives of LDH and MDA increased while the activity of SOD and the mean gray values decreased,p0.01.In treatment groups（GIK group,metoprolol group,concomitant group）,the concomitant group of intracellular Ca2＋ significantly decreased（275±53.6,2.7±1.3）,p0.01.The actives of LDH and MDA decreased while the of activity of SOD and the mean gray values increased,p0.01.Conclusion GIK and metoprolol is effective in A/R myocardial cells with fluorescence staining method.It could significantly decreased the level of intracellular LDH,MDA and Ca2＋ due to A/R and repress effectively expression of c-fos protein.The above mentioned results indicated GIK,metoprolol could stabilize free Ca2＋,against Ca2＋ overload and damage of oxygen free radical during A/R.