L型钙通道参与乳化异氟醚对离体心肌细胞缺氧／复氧损伤的保护作用  

L-type calcium channel participates in the protective effect of emulsified isoflurane on the primary cardiac myocytes hypoxia/reoxygenation injury in neonatal rats

作　　者：陈玉培[1] 闵苏[2] 杨孟昌[1] 张文胜[3] 刘进[3] 徐茜[1] 曹德钧[1] 

机构地区：[1]重庆医科大学附属第二医院麻醉科,重庆400010 [2]重庆医科大学附属第一医院麻醉科,重庆400016 [3]四川大学华西医院麻醉与危重急救教研室,四川成都610041

出　　处：《中国药理学通报》2009年第5期585-589,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30772084)

摘　　要：目的探讨乳化异氟醚(EI)对原代培养心肌细胞缺氧/复氧损伤的保护作用及其与钙通道的关系。方法用原代培养乳鼠离体心肌细胞,建立缺氧/复氧(H/R)损伤模型,随机分为5组(n=8),正常组(N组)、缺氧/复氧组(H/R组)、H/R+脂肪乳组(F组)、H/R+1.68 mmol.L-1EI组(EI组)和H/R+1.68 mmol.L-1EI+10 umol.L-1BayK8644组(EIB组)。各组取细胞培养上清液测定LDH活力与cTnI含量,心肌细胞匀浆后测定SOD活力与MDA含量;用倒置显微镜观察心肌细胞生长特性及形态的变化。钙离子探针Fura-2 AM染色后用流式细胞术检测心肌细胞内钙离子浓度。结果与N组比较,各组的LDH、MDA、cTnI、钙离子浓度均升高(P<0.05),SOD下降(P<0.05);与H/R组比较,F组的SOD下降(P<0.05),LDH、MDA、cTnI、钙离子浓度的差异均无统计学意义(P>0.05);EI组和EIB组的LDH、MDA、cTnI、钙离子浓度均降低(P<0.05),SOD升高(P<0.05)。与F组比较,EI组的LDH、MDA、cTnI、钙离子浓度均下降(P<0.05),SOD升高(P<0.05)。与EI组比较,EIB组的LDH、MDA、cTnI、钙离子浓度均升高,SOD降低(P<0.05)。结论乳化异氟醚对原代培养心肌细胞缺氧/复氧损伤时的保护作用与其抑制L型钙通道,降低细胞内钙超载有关。Aim To discuss the mechanism of protective effect of emulsified isoflurane （EI） on neonatal rat hypoxia/reoxygenation （H/R） cardiac myocytes of primary culture and the association with reduction of intracellular calcium overload. Methods The H/R injury model on the basis of in vitro neonatal rat cardiac myocytes culture was prepared and divided into 5 groups randomly, namely, normal control group （N group） , H/R group, H/R＋ fat emulsion group （ F group）, H/R ＋ 1.68 mmol·L^-1 EI group （EI group）, H/R＋1.68mmol·L^-1 EI ＋10umol·L^-1 Bay K8644 group （EIB group）. The clean liquid on the cellular culture was extracted in each group to mensurate LDH and cellular homogenate was made to mensurate SOD and MDA; the microscope was placed upside down to observe the characteristics of cardiac myocytes growth and changes of its forms; the calcium probe Fura-2 AM was and dyed the flow cytometry was used to inspect the calcium concentration in cardiac myocytes. Results Compared to N group, the LDH, MDA, cTnI and intracelluar Ca^2＋ concentration of other groups all increased （ P 〈 0.05 ） and SOD declined （P 〈 0. 05）. Compared to H/R group, SOD of F group declined （P 〈 0.05 ） , and the differences of the LDH, MDA,cTnI and intracelluar Ca^2＋ concentration had no statistic significance （ P 〉 0.05 ） ; LDH, MDA, cTnI and intracelluar Ca^2＋ concentration of El and EIB group declined （P 〈 0. 05） and SOD increased （P 〈 0. 05 ）. Compared to F group, the LDH, MDA , cTnI and intracelluar Ca^2＋ concentration of EI group declined （P 〈 0.05） and SOD increased （ P 〈 0. 05 ）. Compared to EI group, the LDH, MDA , cTnI and intracelluar Ca2 ＋ concentration of EIB group increased （P 〈 0. 05 ） and SOD declined （P 〈 0.05 ）. Conclusions E1 has the protective effect on the neonatal rat cardiac myocytes with H/R injury, and its mechanism may be associated with blocking L-type calcium channel and reduction of intracellular calcium overload.

关 键 词：乳化异氟醚 原代培养 心肌细胞 缺氧 复氧 钙超载 

分 类 号：R-332[医药卫生] R322.11