参麦注射剂对H_2O_2诱导PC12细胞氧化损伤的保护作用及机制

Protective effect and mechanism of Shenmai injection on oxidative damage in PC12 cells induced by H_2O_2

目的探讨参麦注射剂对H_2O_2诱导PC12细胞氧化损伤的保护作用,并初步探讨其保护机制。方法分别向各给药组加入终浓度2.5、5、10 m L·L-1参麦注射剂,孵育0.5 h后,以H_2O_2(100μmol·L-1)刺激PC12细胞1 h造成的细胞氧化应激损伤模型。之后测定细胞存活率、乳酸脱氢酶(LDH)泄漏率、丙二醛(MDA)生成量和超氧化物歧化酶(SOD)活力,并采用Western blot法测定MAPK信号通路的变化。结果参麦注射剂能明显提高H_2O_2氧化损伤PC12模型细胞的存活率(P<0.01);参麦注射剂(2.5、5、10 m L·L-1)降低模型细胞LDH泄漏率(P<0.01),抑制MDA生成,增加SOD活力(P<0.01)。参麦注射剂可降低由H_2O_2刺激引起的Caspase-3上调,以及MAPK通路上的IKKβ、p38、ERK1的上调,通过MAPK通路保护损伤的PC12细胞。结论参麦注射剂在体外拟神经元工具细胞—PC12细胞上具有抗氧化损伤的作用。

AIM To investigate the effect and possible mechanism of Shenmai injection on oxidative damage of PC12 cells induced by H202. METHODS After administration of 2.5, 5, 10 mL·L^-1 concentration of Shenmai injection to the medium respectively, the medium was incubated for 0.5 h before the stimulation with 100 μmol·L^-1 H2O2＂ PC12 cells were cultured and exposed to 100 μmol·L^-1 H2O2 for 1 h to establish oxidative damage model. The protective effect of Shenmai injection was observed by MTT assay, leakage rate of lactic dehydrogenase （LDH）, malondialdehyde （MDA） generation and superoxide dismutase （SOD） activity determination. Simultaneously, Western blot was subjected to detect expression change of MAPK signaling pathway. RESULTS Compared with the model group, 2.5, 5, 10 mL＂ L-1 concentration of Shenmai injection could increase the survival rate （P 〈 0.01 ）, decrease the LDH leakage rate（ P 〈 0.01 ）, suppress the MDA generation and increase the SOD activity （P 〈 0.01 ）. Western blot results showed that Shen- mai injection could protect the PC12 ceils from oxidative damage via suppressing caspase-3, IKKβ, p38, and ERK1 upregulation regulated by MAPK signaling pathway. CONCLUSION Shenmai injection has a obvious antioxidant effects on PC12 cells in vitro.