丙泊酚对布比卡因诱导的PC12细胞毒性和硫氧还蛋白系统的影响

目的：探讨丙泊酚对布比卡因诱导的PC12细胞毒性的保护作用及内源性硫氧还蛋白（Trx）系统在其中的作用。方法：培养的PC12细胞分成四组，正常对照组、丙泊酚组、布比卡因组、丙泊酚＋布比卡因（PB）组，每组6孔。培养6h和24h后，用MTT比色微量分析细胞存活率，测定上清液乳酸脱氢酶（LDH）活性和细胞内硫氧还蛋白还原酶（TrxR）、活性氧（ROS）活性，RT-PCR检测Trx-1 mRNA和TrxR-1 mRNA表达。结果：与正常PC12细胞相比，布比卡因可显著降低细胞存活率（P〈0．01）和细胞内TrxR活性（P〈0．05），增加上清液中LDH活性和细胞内ROS活性（P〈0．05，P〈0．01），明显降低Trx mRAN和Trx mRAN表达（P〈0．05）；丙泊酚对正常PC12细胞无明显影响；与布比卡因组相比，PB组细胞存活率（P〈0．01）和细胞内Trx活性（P〈0．05）明显增加，上清液中LDH活性和细胞内ROS活性显著降低（P〈0．05，P〈0．01），Trx mRAN和Trx mRAN表达明显增加（P〈0．05）。结论：布比卡因对PC12细胞具有毒性作用可能与降低细胞内TrxR活性、增加ROS活性有关，丙泊酚通过保护细胞内Trx系统的活性及清除ROS来减轻布比卡因诱导的PC12细胞毒性。

Effect of propofol on cytotoxicity and endogenous thioredoxin system induced by bupivacaine in pheochromocytoma PC12 cells

AIM: To investigate protective effect of propofol on the cytotoxicity of bupivacaine in pheochromocytoma PC12 cells of rat and role of endogenous thioredoxin (Trx) system. METHODS: The PC12 cells were inoculated and cultured on the culture capsule. The cultured PC12 cells were randomly assigned to one of four groups (n=6 holes each): Control group, bupivacaine group (B), propofol group (P) and bupivacaine combined with propofol group (BP). Survival rate of PC12 cells (%) treated with different drugs for 6 and 24 h induced was analyzed by MTT assay. Lactate dehydrogenase (LDH) in the culture medium was measured by spectroscopy. Thioredoxin reductase (TrxR) and ROS activities in the PC12 cells were assayed by spectrophotometer. The expression levels of Trx mRNA and TrxR mRNA were determined by RT-PCR. RESULTS: Compared with normal PC12 cells, bupivacaine significantly reduced the relative survival rate (P<0.01) and TrxR activity of the PC12 cells (P<0.05), markedly increased level of LDH in the culture medium(P<0.05) and ROS activity in the PC12 cells (P<0.01), and significantly decreased the expression of Trx mRNA and TrxR mRNA (P<0.05). Compared with B group, the relative survival rate of the PC12 cells were significantly increased(P<0.01) and TrxR activity of the PC12 cells were increased (P<0.05); The level of LDH in the culture medium(P<0.05) and ROS activity in the PC12 cells markedly decreased (P<0.01); The expression of Trx mRNA and TrxR mRNA was significantly increased (P<0.05) in PB group compared with those in B group. CONCLUSION: Bupivacaine induces cytotoxicity in the PC12 cell line, which may be associated with ROS production and decrease in TrxR activity. Propofol can prevent cytotoxicity of PC12 cells induced by bupivacaine, and one of the mechanisms may be associated with protecting endogenous thioredoxin system and scavenging ROS production.