异氟烷对心肌细胞血管内皮生长因子表达的影响

The Effect on the Expression of Vascular Endothelial Growth Factor via Isoflurane in Cardiac Myocytes

目的探讨异氟烷对心肌细胞血管内皮生长因子(VEGF)的直接调节作用及其可能的细胞内信号转导途径,揭示异氟烷心肌保护的分子机制。方法分离培养原代大鼠心肌细胞,随机分为:对照组(CON),细胞不经任何处理;异氟烷组(ISO),细胞分别经0.5、1、1.5MAC的异氟烷处理6h;PKC抑制剂组(CAL),细胞经calphostin C 50 n M孵育6 h;PKC抑制剂+异氟烷组(IC),细胞经calphostin C及异氟烷共同作用6 h。实验结束采用Elisa法测定细胞培养液VEGF浓度。结果异氟烷呈浓度依赖性增加心肌细胞VEGF分泌,与对照组(0MAC)比较,1MAC和1.5MAC组VEGF值均显著升高(P<0.01);PKC抑制剂calphostin C能抑制异氟烷处理引起的VEGF分泌(P<0.01),但calphostin C对心肌细胞VEGF分泌无显著影响(P>0.05)。结论异氟烷诱导心肌细胞释放VEGF,有可能是异氟烷心肌保护机制之一,蛋白激酶C参与异氟烷诱导的VEGF表达。

Objective To investigate the effect of isoflurane on Vascular endothelial growth factor （VECF） expression and associated signaling pathway in cultured rat cardiac myoeytes. Methods Primary cultures of rat eardiomyoeytes were randomized into the following groups： the control group： cells were incubated without any treatment; The isoflurane group： cells were treated with isoflurane at 0.5,1,1.5MAC respectivly for 6 hours ; The PKC inhibitor group ： cells were treated with PKC inhibitor ealphostin C at a final concentration of 50 nM; The PKC inhibitor ＋ isoflurane group： cells were treated with 50nM calphostin C and 1 MAC isoflurane for 6 hours. VEGF expressions were identified by enzyme-linked inmmnosorbent assay. Results Isoflurane dose dependently increased cardiac myocytes VEGF expression. The VEGF levels were significantly higher in 1 MAC and 1.5MAC isoflurane group as compared with the control group （P〈0.01）.The effect of isoflurane on VEGF upregulation was blocked by PKC inhibitor ealphostin C, however,calphostin did not alter VEGF expression （P〉0.05）. Conclusion We demonstrated that isoflurane induced VECF expression in cardiac myocytes through PKC, suggesting a possible novel mechanism of isoflurane induced cardiac protection.