乳化异氟醚对乳鼠原代培养缺氧/复氧损伤心肌细胞保护效应的最适浓度研究

目的 探讨乳化异氟醚(EI)对乳鼠原代培养缺氧/复氧(H/R)损伤心肌细胞保护效应的最适浓度.方法 原代培养乳鼠心肌细胞,建立体外心肌细胞H/R损伤模型,随机分为13组:正常组(N组),缺氧/复氧组(H/R组),脂肪乳组(F组),按0.28 mmol/L EI的1、2、3、4、5、6、7、8、9、10倍分别分成EI1～ EI10组.各组取细胞培养上清液测定乳酸脱氢酶(LDH)活性和心肌肌钙蛋白I(cTnI)含量,细胞匀浆后测定心肌细胞超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量.结果 与N组比较,各组的LDH、MDA、cTnI均升高(P<0.05),SOD下降(P<0.05).与H/R组比较,EI各组的LDH、MDA、cTnI均降低(P<0.05),SOD升高(P<0.05).与EI6组比较,EI其余各组的LDH、MDA、cTnI均升高(P<0.05),SOD下降(P<0.05).随着EI浓度(倍数递增)的增加,EI1~EI6组的LDH、MDA、cTnI逐渐下降,SOD逐渐升高(P<0.05),EI7~EI10组的LDH、MDA、cTnI逐渐升高,SOD逐渐下降(P<0.05).结论 EI对乳鼠离体心肌细胞H/R损伤具有保护作用,其最佳心肌保护效应浓度为1.68 mmol/L,其机制可能与其抗氧化作用有关.

The Optimal Concentration for the Protective Effect of Emulsified Isoflurane on the Neonatalrat Cardiac Myocytes Hypoxia/Reoxygenation Injury of Primary Culture

Objective To optimize the concentration of emulsified isoflurane (El) for the protective effect on primary cultured neonatal rat hypoxia/reoxygenation (H/R) cardiac myocytes. Methods To prepare the H/R injury model on the basis of in-vitro neonatal rat cardiac myocytes culture and divide them into 13 groups at random, namely, normal control group (N group), H/R group, H/R+fat emulsion group (F group) , the one, two, three, four, five, six, seven, eight, nine and ten times of 0. 28 mmol/L El designated as EI1-EI10 group. The supernatants of cell culture from each group were detected for lactate dehydrogenase (LDH) activity and the level of cardiac troponin-I (cTnl). The cellular homogenates of each group were prepared for the detection of superoxide dismutase (SOD) and malondialdehyde (MDA). Inverted microscope was applied to observe the characteristics of cardiac myocytes growth and changes of its forms. Results Compared to N group, the LDH. MDA and cTnl of others all increased (P<0. 05) and SOD decreased (P<0. 05). Compared to H/R group, LDH, MDA and cTnl of each El dose group decreased significantly (P<0. 05), but SOD increased (P<0. 05). Compared to EL. group, the LDH, MDA and cTnl in the other groups of El increased (P<0. 05) and the SOD decreased (P<0. 05). As the El concentration (increasing by multiple) increased, the LDH, MDA and cTnl in the El1 to EI6 group decreased gradually and SOD gradually increased (P<0. 05), the LDH, MDA and cTnl in the EL to EI10 group increased gradually and SOD gradually decreased (P<0. 05). Conclusion El has the protective effect on the neonatal rat cardiac myocytes with H/R injury, the optimal concentration for the protective effect on cardiac myocytes is 1. 68 mmol/L, and its mechanism may be associated with its anti-oxidation effect.