The Anti-apoptotic Effect Of 17?-estradiol On Chondrocytes Induced By IL-1?

Purpose: To research the role of estradiol in interleukin 1?(IL-1?)induced apoptosis of chondrocyte.Methods: Rat chondrocytes were isolated and cultured in vitro.Cells were pretreated with 17-? estradiol and estrogen receptor blockers(ICI182780)respectively,followed by incubation with 50 ng / ml concentration rr IL-1? induced chondrocyte apoptosis.Apoptosis was evaluated by Annexin V-FITC/PI double staining;Cell viability was assessed by CCK-8 assay;The apoptosis related gene bax,bcl-2,mmp3 and mmp13 m RNA expression levels were evaluated by quantitative RT-PCR;MMP3,MMP13 protein expression levels were detected by Western blotting.Result:Cultivation of SD rat chondrocytes appearance shows polygonal mostly,have abundant cytoplasm,cell nucleus is located in the center of the cell body,a few can have multiple nucleoli,the original generation of cartilage cells cultured for 10 days or so to achieve fusion state.Toluidine blue stain make normal cartilage cell cytoplasm is blue,the nuclei are purplish blue,most were round or oval,visible multiple nucleoli.CCK 8 test results found that: compared with control group,IL-1?group,only about 40% of the survival cells decreased significantly.Compared with IL-1?group,17-?E2 group + IL-1? cell survival rate increased significantly,and a dose dependent.IL-1 beta group and 17-?E2 + IL-1?cell survival rate between + ICI group no significant change.Through the Annexin V-FITC and PI double standard beta induced fluorescence staining showed IL-1? set of three groups of different concentration of percentage of chondrocytes apoptosis 17-?E2(10-7 M,10-9 M,10-11 M)+ IL-1? group are high.17-?E2+IL-1?+ICI group compared with IL-1?induced group had no significant difference.Rt-pcr method for detection of apoptosis related gene,compared with control group,IL-1? after induction,cell Bax expression,the Bcl-2 expression is reduced,the difference is statistically significant.17?E2 concentration of three groups all have the inhibitory effect of IL-1?raised Bax expression,and IL-1? groupand 17?E2+IL-1?+ICI group comparative differences are statistically significant.Western blotting test found that group B cells MMP3 and MMP13 / GAPDH is bigger than the grey value of A and C group(P < 0.05),and group D MMP3 and MMP13 / GAPDH is bigger than the grey value of A and C group(P < 0.05).Conclusion: IL-1? could induced apoptosis of rat chondrocytes,which was resisted by 17?-estradiol by down-regulating MMP-3 and MMP-13.Estrogen may have potential protective effect of cartilage.