脂肪乳剂逆转布比卡因心肌毒性中线粒体凋亡途径相关蛋白的变化

目的检测脂肪乳剂逆转布比卡因心肌毒性作用中线粒体凋亡通路相关蛋白的变化,探讨脂肪乳剂逆转布比卡因心脏毒性的可能机制。方法日龄1~3d的健康SD大鼠乳鼠,取其心室肌进行体外原代培养,用含有布比卡因的培养基培养24h来建立心肌细胞布比卡因中毒模型。实验分组:空白对照组(C组)、布比卡因组(B组)、布比卡因+脂肪乳剂组(BL组)。采用流式细胞仪检测细胞凋亡情况,采用Western blot法检测细胞内细胞色素C(cytochrome C,Cyto-C)和活化的casepase-3蛋白含量。结果与C组比较,B组和BL组细胞凋亡率明显升高,且B组明显高于BL组(P0.05)。与C组比较,B组和BL组Cyto-C和活化的casepase-3含量均明显升高(P0.05),且B组明显高于BL组(P0.05)。结论脂肪乳剂可能通过抑制线粒体Cyto-C释放,降低casepase-3激活来调节细胞凋亡,从而发挥保护心肌细胞的作用。

Protein expression changes of mitochondrial apoptotic pathway during the reverse effects of lipid emulsion on bupivacaine cardiotoxicity

Objective To detect the protein expression changes of mitochondrial apoptotic pathway during the reverse effects of lipid emulsion on bupivacaine cardiotoxicity, so as to investigate the probable mechanism concerning the reverse effect of lipid emulsion on bupivacaine cardiotoxicity. Methods The ventricular muscles of 15 healthy SD neonatal mice (1-3 d) were chosen to conduct primary culture in vitro. And the cardiomyocytes were cultivated in a medium containing bupivacaine for 24 hours to establish its bupivacaine poisoning model. The cultured cardiomyocytes were divided into three groups: control group (group C); bupivacaine group (group B); and bupivacaine+lipid emulsion group (group BL). Flow cytometry was applied to examine the apoptosis of cardiomyocytes, and the Western blot was employed to detect the protein expression variation of cytochrome C (Cyto-C) and cleaved casepase-3. Results Compared with group C, the apoptosis rate was remarkably increased in both group B and group BL and that of the group B was dramatically higher than that of the group BL, with a statistical significance (P<0.05). Compared with group C, the protein expression levels of both Cyto-C and cleaved casepase-3 were significantly increased in groups B and BL (P<0.05), and the protein expression levels of both Cyto-C and cleaved casepase-3 in group B were significantly higher than those in group BL (P<0.05). Conclusion Lipid emulsion can regulate apoptosis through inhibiting the release of mitochondrial Cyto-C and reducing casepase-3 activation, thus it protects cardiomyocytes.