Effects Of TanshinoneⅡA On Oxidative Stress And Apoptosis Cytokine Expression Of Human Peritoneal Mesothelial Cells

Background and ObjectiveOxidative stress increase apoptosis factor expression that can start the occurrenceof cell apoptosis which play an important role in tissue fibrosis in vivo,includingperitoneal fibrosis.Peritoneal fibrosis is a major cause of long-term peritoneal dialysisexit treatment,but peritoneal mesothelial cells apoptosis is a key link in the process ofinitiating lead to fibrosis.To investigate the effect of tanshinoneⅡ A （TSN） onoxidative stress and apoptosis of human peritoneal mesothelial cells （HPMCs）stimulated by peritoneal dialysis fluid （PDF）.MethodsHPMCs were cultured in vitro from HPMCs cell lines. All experiment wereperformed using cells between the first to third passages. Morphologic changes weredetected through downward microscope during cell culture.Oxidative stress indexdetection: HPMCs cultured in vitro were divided into control group （DMEMmedium）, PDF group （4.25%PDF）, TSN group （DMEM medium with100μmol/LTSN）, TSN low concentration group （4.25%PDF with50μmol/L TSN）, TSN highconcentration group （4.25%PDF with100μmol/L TSN）. After48hours the levels ofMDA, SOD and GSH in the supernatant were detected.Apoptosis cytokines detection:HPMCs cultured in vitro were divided into control group （DMEM medium）, H₂O₂group （3%H₂O₂）, TSN+H₂O₂group （3%H₂O₂with50μmol/L TSN）, PDF group（4.25%PDF）, TSN+PDF （4.25%PDF with50μmol/L TSN）. After72hours oftreatment, apoptosis cytokines bax, caspase-3and inhibition apoptosis cytokines bcl-2mRNA and protein were detected by Real time PCR and Western Blot, respectively.Result Compared with PDF group, groups with TSN significantly decreased the level ofMDA and increased the level of SOD and GSH. However, there is no significantdifference between the two different concentration of TSN groups.Compared withPDF and H₂O₂groups, PDF and H₂O₂with TSN significantly decreased theexpression of both bax and caspase-3mRNA and protein and increased the expressionof bcl-2mRNA and protein.ConclusionTSN may down-regulate PDF induced high expression of apoptosis cytokinesthrough inhibiting oxidative stress of HPMCs, which delay the apoptosis of HPMCsand save HPMCs, and thus has a resistance to fibrosis effect on peritoneal membrane.