Salvinorin A对原代培养的大鼠皮层神经元缺氧无糖损伤的保护作用及其机制

Neuroprotective effect and mechanism of Salvinorin A on the injury of oxygen-glucose deprivation in cortical neurons of rats by primary culture

目的研究Salvinorin A对原代培养的大鼠皮层神经元缺氧无糖(OGD)损伤的保护作用及其可能的机制。方法原代培养Sprague-Dawley胎鼠原代皮层神经元。实验分为两个部分,第一部分检测Salvinorin A是否具有神经保护效应,将培养的神经元分为正常对照组、单纯Salvinorin A组(只加入终浓度为10μmol/L的Salvinorin A,不做OGD)、OGD组和OGD加不同浓度Salvinorin A组(0.1μmol/L组、0.5μmol/L组、1μmol/L组、5μmol/L组、10μmol/L组),检测各组的乳酸脱氢酶(LDH)漏出率。第二部分检测Salvinorin A的神经保护机制是通过丝裂原活化蛋白激酶(MAPK)3条途径中的何种途径发挥作用,将原代神经元分为正常对照组、OGD组、OGD+Salvinorin A组(OGD+SA组)、抑制剂+OGD+Salvinorin A组[U0126+OGD+SA组、SB200235+OGD+SA组、SP600125+OGD+SA组,U0126为细胞外信号调节蛋白激酶(ERK)通路抑制剂、SB200235为P38通路抑制剂,SP600125为c-jun氨基末端激酶通路抑制剂],检测各组的LDH漏出率。结果第一部分实验:正常对照组的LDH漏出率与单纯Salvinorin A组的差异无统计学意义(P>0.05),OGD组、0.1μmol/L组、0.5μmol/L组、1μmol/L组、5μmol/L组、10μmol/L组的LDH漏出率均显著高于正常对照组(P值均<0.05),1μmol/L组、5μmol/L组、10μmol/L组的LDH漏出率均显著低于OGD组(P值均<0.05),0.1μmol/L组、0.5μmol/L组的LDH漏出率与OGD组的差异均无统计学意义(P值均>0.05)。第二部分实验:U0126+OGD+SA组的LDH漏出率与OGD组的差异无统计学意义(P>0.05),但显著高于OGD+SA组(P<0.05);SB200235+OGD+SA组、SP600125+OGD+SA组的LDH漏出率均显著低于OGD组(P值均<0.05),但与OGD+SA组的差异均无统计学意义(P值均>0.05)。结论 Salvinorin A对大鼠原代神经元OGD损伤具有保护作用,其机制与MAPK/ERK通路有关。

Objective To investigate the neuroprotective effects and mechanisms of Salvinorin A on the injury of oxygen-glucose deprivation （OGD） in primary cultured cortical neurons of rats. Methods Primary culture was performed for cortex neurons of Sprague-Dawley fetal rats. The experiment had two parts. The first part was tO investigate whether Salvinorin A had neuroprotective effects. In this part, neurons were divided into control group, SA group （10 μmol/L Salvinorin A was given）, OGD group, and OGD＋ Salvinorin A （0.1, 0.5, 1, 5, 10 μmol/L） groups. The lactate dehydrogenase （LDH） release rate was measured in each group. The second part was to investigate which signal pathway of mitogen-activated protein kinase （MAPK） was involved in the neuroprotection of Salvinorin A. In this part, neurons were divided into control group, OGD group, OGD-I- Salvinorin A group, and inhibitor ＋ Salvinorin A （U0126 ＋ OGD ＋ SA, SB200235 ＋ OGD ＋ SA, SP600125 ＋ OGD ＋ SA, U0126 was the inhibitor of extracellular signal regulating protein kinase[ERK] pathway, SB200235 was P38 pathway inhibitor and SP600125 was c-jun amino terminal kinase pathway inhibitor） groups. The LDH release ratewas also measured in each group. Results The first part= there was no significant difference in the LDH release rate between control group and SA group （P〈0.05） ; the LDH release rates of OGD group and OGD-t- Salvinorin A （0.1, 0.5, 1, 5, 10 μmol/L） groups were significantly higher than that of control group （all P〈0.05） ; the LDH release rates of OGD＋ Salvinorin A （1, 5, 10 umol/L） groups were significantly lower than that of OGD group （all P〈0.05） ; the LDH release rates of OGD ＋ Salvinorin A （0. 1 and 0.5μmol/L） groups were not statistically different from that of OGD group （both P〈0.05）. The second part： the LDH release rate of U0126 ＋ OGD＋ SA group was not statistically different from that of OGD group （ P〈0.05）, but significantly higher than that of OGD＋ SA group （P〈0.05）; the LDH release rate of SB200235 ＋ OGD＋ SA group and SP600125 ＋ OGD ＋ SA group were significantly lower than that of OGD group （both P〈0.05）, but not statistically different from that of OGD＋ SA group （both P〈0.05）. Conclusion Salvinorin A has neuroprotection effect on the OGD injury in primary cultured cortical neurons of rats. ERK/MAPK pathway may be involved in the procedure.