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目的探讨大黄酚对氨诱导小鼠星形胶质细胞的影响及相关机制。方法原代小鼠星形胶质细胞共分5组,分别同时加入氯化铵5 mmol·L~(-1)+大黄酚0.1,1.0和10.0 mg·L~(-1)(共3组),氯化铵5 mmol·L~(-1)+细胞外信号调节激酶1/2(ERK1/2)抑制剂UO126(10μmol·L~(-1))组和氯化铵5 mmol·L~(-1)+p38丝裂原活化蛋白激酶(p38 MAPK)抑制剂SB239063(10μmol·L~(-1))组,处理48 h。紫外分光法测定氧化应激指标丙二醛(MDA)、一氧化氮(NO)的含量以及谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)和一氧化氮合酶(NOS)的活性;Western蛋白印迹法检测ERK1/2和p38 MAPK磷酸化水平,RT-PCR检测c-fos和c-jun m RNA的表达。结果与氯化铵5 mmol·L~(-1)相比,大黄酚1.0和10.0 mg·L~(-1)显著改善氨诱导的细胞氧化应激,降低了MDA和NO的含量及NOS的活性(P<0.05),明显升高了GSH-Px和SOD的活性(P<0.05);大黄酚1.0和10.0 mg·L~(-1)也明显降低氨诱导ERK1/2和p38 MAPK的磷酸化增加(P<0.05);大黄酚0.1,1.0和10.0 mg·L~(-1),UO126和SB239063均可逆转氨诱导c-fos和c-jun m RNA表达下调的作用(P<0.05)。结论大黄酚通过抗氧化机制影响ERK1/2和p38 MAPK的磷酸化,进而逆转氨诱导c-fos和c-jun m RNA表达下调。
Objective To investigate the effect of chrysophanol on astrocyte induced by ammonia and its related mechanism. Methods The primary mouse astrocytes were divided into 5 groups. At the same time, 5 mmol·L -1 chitosan 0.1,1.0 and 10.0 mg · L -1 ammonium chloride ), 5 mmol·L -1 ammonium chloride (5 mmol·L -1) extracellular signal-regulated kinase 1/2 (ERK 1/2) inhibitor UO126 (10 μmol·L -1) ~ (-1) + p38 mitogen activated protein kinase (p38 MAPK) inhibitor SB239063 (10μmol·L -1) for 48 h. The content of malondialdehyde (MDA), nitric oxide (NO) and the content of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and nitric oxide (NOS). The phosphorylation levels of ERK1 / 2 and p38 MAPK were detected by Western blotting. The expressions of c-fos and c-jun mRNA were detected by RT-PCR. Results Compared with 5 mmol·L -1 ammonium chloride, 1.0 and 10.0 mg · L -1 of chrysophanol could significantly reduce the oxidative stress induced by ammonia, decrease the content of MDA and NO, (P <0.05), significantly increased the activities of GSH-Px and SOD (P <0.05), while 1.0 and 10.0 mg · L -1 of chrysophanol significantly decreased the phosphorylation of ERK1 / 2 and p38 MAPK (P <0.05). Chrysophanol 0.1, 1.0 and 10.0 mg · L -1, UO126 and SB239063 all could reverse the effect of ammonia-induced down-regulation of c-fos and c-jun m RNA expression . Conclusion Chrysophanol can affect the phosphorylation of ERK1 / 2 and p38 MAPK through anti-oxidative mechanism, and then reverse the down-regulation of ammonia-induced c-fos and c-jun m RNA expression.