World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(3):486-490

doi:10.11842/wst.2017.03.017

Protective Effects of Zhen-Gan Xi-Feng Decoction-containing Serum on 6-OHDA-induced Oxidative Stress in PC12 Cells through Nrf2-ARE Pathway Activation

Xuemei ZHAO ; Tianjiao XU ; Miaoxian DONG

Keywords

Zhen-Gan Xi-Feng decoction; 6-hydroxydopamine; reactive oxygen species; Nfe2l2; antioxidant response element; drug-containing serum

Country

China

Language

Chinese

Abstract

This paper was aimed to study the protective effects and related mechanisms of Zhen-Gan Xi-Feng (ZGXF) decoction containing serum on 6-OHDA-induced oxidative stress in PC12 cells.The ZGXF decoction containing rat serum with low-,medium-,and high-dose (8,16,or 32 g.kg-1) or blank serum was used to preprocess PC12 cells for 1 h,and cultured together with 100 μM 6-OHDA for 24 h.And then,cells were collected.The fluorescent probe 2',7'-dichlorofluorescin diacetate (DCFH-DA) and fluorescence microplate reader were used to detect the level of reactive oxygen species (ROS).Real-time quantitative PCR was used to analyze the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2),Nfe2l2,heme oxygenase-1 (HO-1),glutamate-cysteine ligase catalytic subunit (GCLc),and GCL modulatory subunit (GCLm).The luciferase report gene system was used to detect the antioxidant response element (ARE) activation.The results showed that ZGXF decoction-containing serum inhibited the 6-OHDA-induced oxidative stress,upregulated the Nfe2l2,HO-1 and GCLc mRNA expressions in cells processed with 6-OHDA.However,it has no significant effect on GCLm mRNA expression.It was concluded that ZGXF decoction-containing serum had protective effects on 6-OHDA-induced oxidative stress in PC 12 cells.Its mechanism may be correlated with the upregulation on Nfe2l2 mRNA expression,which activated ARE and further induced its downstream gene of phase Ⅱ detoxifying enzyme,as well as the HO-1 and GCLc mRNA expressions of antioxidant enzyme gene.