镇肝熄风汤含药血清对6-OHDA诱导PC12细胞损伤保护作用的抗氧化机制分析  

Antioxidant Mechanism of Zhengan Xifeng Tang-containing Serum in Protecting 6-OHDA-induced PC12 Cell Injury

作　　者：邹宇[1] 董妙先[2] 

机构地区：[1]齐齐哈尔医学院药学院,黑龙江齐齐哈尔161006 [2]齐齐哈尔医学院医药科学研究所,黑龙江齐齐哈尔161006

出　　处：《中国实验方剂学杂志》2017年第9期112-116,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81373629)

摘　　要：目的:观察镇肝熄风汤含药血清对6-羟基多巴胺(6-hydroxydopamine,6-OHDA)诱导PC12细胞凋亡的影响,并探讨其是否通过抗氧化活性发挥神经保护作用。方法:将40只Wistar大鼠随机分为空白组、镇肝熄风汤低、中、高剂量组,每组10只。镇肝熄风汤按照8,16,32 g·kg^(-1)灌胃,空白组灌胃等体积生理盐水,采血制备含药和空白血清。常规培养PC12细胞,分为空白组、模型组、镇肝熄风汤低、中、高剂量组,空白组和模型组给予空白血清,其他3组给予10%含药血清,孵育1 h,再加100μmol·L^(-1)6-OHDA共孵育24 h后收集细胞。采用MTS法检测细胞活性,流式细胞术检测细胞凋亡,分光光度法检测脂质过氧化物(lipid peroxidation,LPO)水平,实时荧光定量PCR(Real-time PCR)检测醌氧化还原酶-1(NADPH quinine oxidoreductase 1,NQO-1)mRNA表达。结果:与空白组比较,模型组细胞活力明显降低,而细胞凋亡和LPO水平明显增加(P<0.05)。与模型组比较,10%的镇肝熄风汤低、中、高剂量含药血清组细胞活力显著增加,而细胞凋亡和LPO水平明显减少(P<0.05)。与模型组比较,10%的镇肝熄风汤低、中、高剂量含药血清组NQO-1 mRNA表达明显上调(P<0.05),而空白组与模型组NQO-1mRNA表达无显著性差异。结论:镇肝熄风汤含药血清具有保护6-OHDA诱导PC12细胞损伤的作用,其机制可能与上调PC12细胞NQO-1 mRNA表达进了提高了细胞抗氧化能力有关。Objective： To examine the effect of Zhengan Xifeng Tang on 6-hydroxydopamine （6-OHDA）-induced apoptosis of PC12 cell, and its antioxidant activity on neuroprotection. Method： A total of 40 male Wistar rats were randomly divided into the control group and high, middle and low-dose Zhengan Xifeng Tang groups （32, 16, 8 g·kg^-1）. Rats were given Zhengan Xifeng Tang with intragastric administration to prepare Zhengan Xifeng Tang-containing serum. The control group was given the same volume of normal saline. The proliferation of PC12 cells were cultured and divided into the blank group, the model group and high, middle and low-dose Zhengan Xifeng Tang groups. The blank group and the model group were given blank serum, the other three groups were given 10% drug-containing serum incubated for 1 h, and added with 100 μmol·L^-1 6-OHDA to co-cultured for 24 h. the cell viability was measured by MTS. Apoptosis was analyzed by flow cytometry. Measurements of lipid peroxidation （LPO） levels were performed using spectrophotometry method. Real-time PCR were used to analyze the expression of NADPH quinine oxidoreductase 1 （NQO-1） mRNA. Result： Compared with blank control, cell viability was significantly decreased, whereas apoptosis and LPO levels were significantly increased in 6-OHDA-exposed PC12 cells （P〈0.05）. Compared with the model group, cell viability were significantly increased, whereas apoptosis and LPO levels were significantly decreased in Zhengan Xifeng Tang-containing serum treatment groups （P〈0.05）. Compared with the model group, NQO-1 mRNA expression were significantly up-regulated in Zhengan Xifeng Tang-containing serum treatment groups （P〈0.05）, but with no significant difference in NQO-1 mRNA expression between the normal group and model group. Conclusion： Zhengan Xifeng Tang-containing serum has the neuroprotective effect against 6-OHDA-induced apoptosis, which may be attributable to its antioxidant properties through up-regulation of NQO-1 mRNA expression.

关 键 词：镇肝熄风汤 血清药理 神经保护 细胞凋亡 氧化应激 醌氧化还原酶-1 脂质过氧化物 

分 类 号：R285.5[医药卫生—中药学]