Abstract Objective:To investigate whether hPTH1-34 regulate the synthesis of fibronectin(FN) from cul-trued rat mesangial cells and its possible mechanism.Methods:(1) MCs seeded at a density of 1×10^4 per well in...Abstract Objective:To investigate whether hPTH1-34 regulate the synthesis of fibronectin(FN) from cul-trued rat mesangial cells and its possible mechanism.Methods:(1) MCs seeded at a density of 1×10^4 per well in 24-well plates were treated with medium containing various concentrations of hPTH1-34(10^-12mol/l-10^-8mol/l)for 6h,12h,24h and 48h,control cells were treated with vehicle only.The FN levels (in the supernatant)were measured by ELISA assay.(2) MCs were co-cultured with 10ng/l of anti-TGF-βanti-body and various concentrations of hPTH1-34(10^-12mol/l-10^-6mol/l).Forty-eight hours later, FN were tested by ELISA.(4)MCs were co-cultured with 10ng/l of anti-TGF-β antibody and 10^-8 mol/l hPTH1-34 for 6h,12h,24h and 48h and then FN were tested.Results(1)hPTH1-34 stimulated FN synthesis in a dose-and time-dependent way with a peak at 10^-8 mol/l(P<0.01).(2)Anti-TGF-β antibody inhibited the stimu-lation effect of hPTH1-34 on synthesis of FN in cultured rat mesangial cells(P<0.05).Conclusion:hPTH1-34 up-regulates FN synthesis in cultured rat mesangial cells via TGF-β ,suggesting that PTH may play an im-portant role in deteriorating the residual renal function at the early stage of chroic renal disease.展开更多
Objective To investigate the role of cyclin-kinase inhibitor p27 on proliferation of mesangial cell(MC) induced by tumor necrosis factor α( TNF-α ). Methods The p27 protein of MC lysate was detected with western blo...Objective To investigate the role of cyclin-kinase inhibitor p27 on proliferation of mesangial cell(MC) induced by tumor necrosis factor α( TNF-α ). Methods The p27 protein of MC lysate was detected with western blotting analysis. The degree of MC proliferation was estimated through [^3H] thymidine incorporation. The effect of reducing p27 expression on MC proliferation was analysed with p27 antisense oligodeoxynucleotide (ODN). Results TNF-α(200000U/L) decreased p27 level to (0.6±0.1 ) from (1. 1±0.1 ) of MC lysate cultured in serum free DMEM for 24h ( P<0.01 ) and increased [^3H] thymidine incorporation to ( 2060±112 ) from (685±53) cpm/well( P<0.01 ). p27 antisense ODN transfection decreased p27 level of MC stimulated by TNF-α for 24h [(0.3±0.1 ) vs (0.6±0.1), P <0.01 ] and increased [^3H] thymidine incorporation [(2420±130) vs (2060±112) cpm/well, P <0.05]. Conclusion The decline of p27 protein maybe play an important role in MC proliferation induced by TNF-α.展开更多
文摘Abstract Objective:To investigate whether hPTH1-34 regulate the synthesis of fibronectin(FN) from cul-trued rat mesangial cells and its possible mechanism.Methods:(1) MCs seeded at a density of 1×10^4 per well in 24-well plates were treated with medium containing various concentrations of hPTH1-34(10^-12mol/l-10^-8mol/l)for 6h,12h,24h and 48h,control cells were treated with vehicle only.The FN levels (in the supernatant)were measured by ELISA assay.(2) MCs were co-cultured with 10ng/l of anti-TGF-βanti-body and various concentrations of hPTH1-34(10^-12mol/l-10^-6mol/l).Forty-eight hours later, FN were tested by ELISA.(4)MCs were co-cultured with 10ng/l of anti-TGF-β antibody and 10^-8 mol/l hPTH1-34 for 6h,12h,24h and 48h and then FN were tested.Results(1)hPTH1-34 stimulated FN synthesis in a dose-and time-dependent way with a peak at 10^-8 mol/l(P<0.01).(2)Anti-TGF-β antibody inhibited the stimu-lation effect of hPTH1-34 on synthesis of FN in cultured rat mesangial cells(P<0.05).Conclusion:hPTH1-34 up-regulates FN synthesis in cultured rat mesangial cells via TGF-β ,suggesting that PTH may play an im-portant role in deteriorating the residual renal function at the early stage of chroic renal disease.
文摘Objective To investigate the role of cyclin-kinase inhibitor p27 on proliferation of mesangial cell(MC) induced by tumor necrosis factor α( TNF-α ). Methods The p27 protein of MC lysate was detected with western blotting analysis. The degree of MC proliferation was estimated through [^3H] thymidine incorporation. The effect of reducing p27 expression on MC proliferation was analysed with p27 antisense oligodeoxynucleotide (ODN). Results TNF-α(200000U/L) decreased p27 level to (0.6±0.1 ) from (1. 1±0.1 ) of MC lysate cultured in serum free DMEM for 24h ( P<0.01 ) and increased [^3H] thymidine incorporation to ( 2060±112 ) from (685±53) cpm/well( P<0.01 ). p27 antisense ODN transfection decreased p27 level of MC stimulated by TNF-α for 24h [(0.3±0.1 ) vs (0.6±0.1), P <0.01 ] and increased [^3H] thymidine incorporation [(2420±130) vs (2060±112) cpm/well, P <0.05]. Conclusion The decline of p27 protein maybe play an important role in MC proliferation induced by TNF-α.
