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.
ABSTRACT
The aim of this study was to explore the effects of ZGXF decoction on amphetamine-induced rotation in PD rats with syndrome of liver-yang hyperactivity,and its mechanism involved.Rats received 6-OHDA administration via intra-substantia nigra injection and were intragastrically treated by Fu Zi decoction to establish the PD model with the syndrome of liver-yang hyperactivity.Three doses of ZGXF decoction or selegiline were given by a 28-day intragastric administration.The rats were tested for amphetamine-induced rotation asymmetry.In addition,real-time PCR were adopted to analyze the expressions of Nfe212 and Hmox1 mRNAs,while western blot to analyze the expression of Keap1 protein.As a result,it was found that ZGXF decoction dose-dependently attenuated amphetamine-induced rotation,up-regulated the expressions of Nfe212 and Hmox-1 mRNAs,and down-regulated the expression of Keap1 protein in the substantia nigra in PD rats with syndrome of liver-yang hyperactivity.It was suggested that anti-PD effects of ZGXF decoction be attributed to the up-regulation of Nfe212 and Hmox-1 mRNAs and the down-regulation of Keap1 protein,being associated with oxidative stress,in the substantia nigra of PD rats with syndrome of liver-yang hyperactivity.