p38丝裂原活化蛋白激酶在罗哌卡因致SH—SY5Y细胞凋亡中的作用

Role of p38MAPK in ropivacaine-induced SH-SY5Y cell apoptosis

目的评价p38丝裂原活化蛋白激酶（p38MAPK）在罗哌卡因致SH—SY5Y细胞凋亡中的作用，以探讨罗哌卡因诱发神经毒性的机制。方法采用随机数字表法，将SH—SY5Y细胞随机分为4组（n=18）：正常对照组（c组）、10μmol／Lp38MAPK特异性抑制剂SB203580组（sB组）、3mmol／L罗哌卡因组（R组）、10μmol／LSB203580＋3mmol／L罗哌卡因组（sB＋R组）。c组在细胞培养液中继续培养；SB组在含10μmol／LSB203580培养液中孵育；R组在含3mmol／L罗哌卡因的培养液中孵育；SB＋R组在含10μmol／LSB203580的培养液中孵育30min后，用含3mmol／L罗哌卡因的培养液继续孵育。各组细胞培养或罗哌卡因孵育4h后采用流式细胞仪检测细胞内活性氧（ROS）水平和细胞凋亡率，采用Western blot法检测p38MAPK和磷酸化p38MAPK（p-p38MAPK）的表达，采用MTT法检测细胞活力。结果与c组比较，R组和sB＋R组ROS水平升高，p-p38MAPK表达上调，细胞活力降低，细胞凋亡率升高（P〈0．01），sB组上述指标差异无统计学意义（P〉0．05）；与R组比较，sB组p-p38MAPK表达下调，细胞活力升高，细胞凋亡率降低（P〈0．01）。四组p38MAPK表达水平差异无统计学意义（P〉0．05）。结论罗哌卡因致SH—SY5Y细胞凋亡作用的机制部分与p38MAPK的激活有关。

Objective To evaluate the role of p38MAPK in ropivacaine-induced SH-SY5Y cell apoptosis and investigate the mechanism by which ropivacaine induced neurotoxicity. Methods SH-SY5Y cells were ran-domly divided into 4 groups （ n = 18 each） ： control group （group C）, p38MAPK specific inhibitor SB203580 10 μmol/L group （group SB）, ropivacaine 3 mmol/L group （group R）, and SB203580 10 p_mol/L ＋ ropivacaine 3 mmol/L group （group SB ＋ R） . The cells were continuously cultured in the culture medium in group C. The cells were incubated in the culture medium containing SB203580 10μmol/L and ropivaeaine 3 mmol/L in groups SB and R respectively. The cells were incubated in the culture medium containing SB203580D 10 μmol/L for 30 rain and then in the culture medium containing ropivaeaine 3 mmol/L in group SB ＋ R. After the cells were cultured or incubated with ropivacaine for 4 h, the intracellular reactive oxygen species（ROS） level and apoptosis rate were detected by flow cytometry, the expression of p38MAPK and phospho-p38MAPK （p-p38MAPK） was determined by Western blot, and the cell viability was detected by MTT assay. Results Compared with group C, the ROS level was significantly increased, the expression of p-p38MAPK up-regulated, the cell viability decreased, and the apoptosis rate increased in groups R and SB ＋ R （P 〈 0.01 ）, while no significant change in the parameters mentioned above was found in group SB （P 〉 0.05）. Compared with group R, the expression of p-p38MAPK was down-regulated, the cell viability was significantly increased, and the apoptosis rate decreased in group SB （P 〈 0.01）. There was no significant difference in the expression of p38MAPK among the four groups （P 〉 0.05）.Conclusion The mechanism by which ropivacaine induces SH-SY5Y cell apoptosis is related to the activation of p38MAPK.