Experimental Study On Oxidative Stress And Mitochondrial Damage Induced By Na-PCP In L-02 Hepatocytes

Objective:To study the mitochondrial damage and the oxidative stress induced by pentachlorophenol sodium(Na-PCP)in L-02 hepatocytes,to build an experiment foundation for better understanding the toxicity of pentachlorophenol sodium(Na-PCP).Methods:With the method of MTT test,the viability of L-02 hepatocytes after treatment with different concentrations of Na-PCP was detected.On the basis,six groups were designed at the concentration of Na-PCP,namely 0mg/L,12.812mg/L,19.218mg/L,25.624mg/L,32.030mg/L and 38.436mg/L;the activity of SOD and the levels of GSH and MDA in L-02 hepatocytes after treatment with Na-PCP for 24 hours were determined by the colorimetric method.The changes of mitochondrial transmembrane potential(△ψm)and permeability pore(PTP)caused by Na-PCP were detected by the fluorospectrophotometric method. Concentrations of 9.609mg/L,12.812mg/L and 19.218mg/L were selected to assay the protein levels of p53,cytochrome C(cytC)and apoptosis-inducing factor(AIF)in L-02 hepatocytes induced by Na-PCP with the method of Western Blotting.Results:1.From 9.609mg/L to 38.436mg/L,Na-PCP significantly decreased the viability of L-02 hepatecytes(r=-0.978,P＜0.01).2.From 12.812mg/L to 35.436mg/L,Na-PCP significantly increased the level of MDA(r=0.956,P＜0.01),decreased the activity of SOD(r=-0.900,P＜0.01)and reduced the level of GSH(r=-0.975,P＜0.01) in L-G2 hepatocytes,by comparing with the control group.3.Compared with the control group,the open rate of PTP was enhanced with the concentration of Na-PCP increasing from 12.812mg/L to 38.436mg/L(P＜0.01),and it showed a concentration-dependent relationship(r=0.997,P＜0.01).4.With the increase of Na-PCP concentration from 12.812mg/L to 38.436mg/L(P＜0.01),mitochondrial transmembrane potential significantly decreased,compared with the control group.And a concentration-response relationship was observed(r=-0.795,P＜0.01).5.By comparing with the control group,with the increase of concentration of Na-PCP from 9.609mg/L to 19.218mg/L,the expression level of p53 protein increased,and that of cytC and AIF in L-02 hepatocytes increased as well.Conclusions:By exposing to a concentration range from 9.609mg/L to 38.436mg/L of Na-PCP,the viability of L-02 hepatocytes was significantly decreased;and the activity of SOD was significantly reduced,the level of GSH was reduced,while the level of MDA was enhanced.Na-PCP induced the collapse of mitochondrial transmembrane potential and increased the open rate of permeability transition pore,and more cytchrome C and AIF were released from mitochondria.Na-PCP increased the protein level of p53 in L-02 hepatocytes as well.