| ObjectivePeroxisome proliferator-activated receptor-gamma(PPAR-γ)is a kind of the most extensive study nuclear hormone receptor,which participates in many kinds of biological effect,PPARγnot only relates to lipid metabolism redox state,inflammation, cardiovascular disease,diabetes,obesity,but also affects cell growth,differentiation and apoptosis.PPARγbelong to nuclear transcription factor,including crude ligands: 15d-PGJ2,linolic acid,fatty acid,oxidized form of low denty lipid ramification and synthetical ligand:anti-diabetic drug Thiazolidinediones(TZDs).The research shows: Activitied PPARTby its ligand can promote many kinds of cells apoptosis in transcription level.Cysteine-aspartate-specific protease(caspase protease)family hold in the network status of the centre molecular mechanism of apoptosis,which caspase-3 protease cascade in the apoptotic process of cutting a core position,and is regarded as apoptosis eventual implementation,a "killer protein".The purpose of this study is to analyse different concentrations of PPARγligands on the JEG-3 cells in the expression of caspase-3,and to explore JEG-3 cells apoptosis induced.by PPARγligands.MethodsJEG-3 cell lines cultured:JEG-3 cells grows in FD(F12:DMEM= 1:1)solution:containing 10%fetal bovine serum,penicillin 100U/ml and streptomycin 100μg/ml,the solution was changed when it became yellow fluid,passaged by 1:2.Admission index JEG-3 cells were growed experiment.Experimental group control group:conventional cell culture and subculture,not 15-d-PGJ2.Treatment groups:The inoculated cells came into logarithmic growth phase after 24 h,in part by adding six orifice,the pretreatment glass coverslips under six orifice,To be 80%of cell crawling, the 10μmol/l 15-d-PGJ2was added into fulid respectivly,applicated in the immunofluorescence experiments after 24 hours,the Another part of accession (0.1,1,10μmol/l)15-d-PGJ2,will be collected after 24 hours,then centrifugated.the EP loaded to prepare for Western Blot test application.ResultsImmunofluorescence experiments:observed under fluorescence microscope to the green fluorescent signal which is caspase-3-positive signal.The results showed that,the green fluorescent signals mainly located in the cytoplasm,suggesting that the main caspase-3 expression in the cytoplasm.JEG-3 cells(0μmol/l)which did not deal with 15-d-PGJ2 performance that scattered the green fluorescent signals in the cytoplasm, signal strength weak,which suggested that only weak caspase-3 expression;and the JEG-3 cells which dealed with 15-d-PGJ2(10μmol/l)for 24 h,though there was no obvious change in appearance,but in the cytoplasm of green fluorescent signal-intensive,and showed the bright green fluorescence signal,signal-intensity, suggesting that by 15-d-PGJ2 cells after treatment of caspase-3 expression levels were significantly elevated.Western blot technology:The results showed that the control group that JEG-3 cells without 15-d-PGJ2 processing(0μmol /l),with the relative expression level of 0.23±0.06,suggesting that intracellular expression of caspase-3 fewer;And the 15-d-PGJ2 cells after treatment with relative expression levels more, suggesting that JEG-3 cells treated with 15-d-PGJ2,the expression of caspase-3 increased.The results also suggest that the 15-d-PGJ2(0.1,1,10μmol/l)treated with 15-d-PGJ2 concentration increased,the relative expression level of the band has gradually increased,0.36±0.03,0.53±0.06,0.71±0.04 respectivly.The JEG-3 cells treated by 0.1μmol/l 15-d-PGJ2 compared with the control group in the expression of caspase-3,has statistically significant difference(P＜0.05),while the concentration of 1.0 and 10μmol/l,compared with the control group,has also statistically significant difference(P＜0.01),suggesting that the JEG-3 cells treated by different concentrations 15-d-PGJ2,the expression of caspase-3 significantly increased,and with 15-d-PGJ2 concentration of the increase in JEG-3 cells,the expression of caspase-3 has also increased,there was dose-dependent relationship each other.Conclusions(1)JEG-3 cells have caspase-3 expression.(2)PPARγligands that 15-d-PGJ2 can induce the expression of caspase-3 increased in JEG-3 cells.(3)with 15-d-PGJ2 concentration of the increase in JEG-3 cells,the expression of caspase-3 has also increased,there is dose-dependent relationship each other,suggesting that PPARγligands can induce JEG-3 cell apoptosis.
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