Effect Of Uric Acid And Urea On HK-2 Cells Apoptosis And Expression Of Fibrotic Correlation Factors And Intervention Of PGE1

Part 1 Effect of uric acid on HK-2 cells apoptosis and expression of fibrotic correlation factors and intervention of PGE1Objective:The study was designed to investigate the effect of UA on renal proximal tubular cells apoptosis and expression of fibrotic correlation factors as well as the protective effect of PGE1 against Urea-induced apoptosis and expression of fibrotic correlation factors in renal proximal tubular cells。Methods:1. HK-2 cells were used as the subject;2.The toxic effect of UA on HK-2 cells was determined by MTT assay,NAG activity and Annexin V.FITC/PI fluorescence.3.Western Blot and Real-Time PCR were employed to investigate the expression of TGF-β1, CTGF and FN. In the blocking study, PGE1 was used as antagonists respectively.Results:HK-2 Cells were exposed to UA (100,200,400, 800umol/L) respectively for 24h,48h,72h. UA could induce a significant dose and time-dependent loss of cell viability. Have been treated with different concentrations UA for 24h, NAG concentration and apoptosis ratio were increased dose dependently (P<0.01). The presence of PGE1 significantly lowered the inhibition rate of cell activity, NAG concentration and apoptotic ratio compared with the cells which were treated with Urea only.The protein and mRNA of TGF-β1,CTGF,FN in HK-2 cells increased significantly when HK-2 cells were cultivated with Urea, when PGE1 was added in UA fibrotic correlation factors decrease significantly.Conclusion:1.UA could inhibition of renal proximal tubular cells proliferation and promote apoptosis in vitro; 2.UA could promote the expression of fibrosis relation factor(TGF-(31,CTGF,FN)in renal tubular epithelial cells; 3. PGE1 can interrupt apoptosis and the expression of fibrosis correlation factor caused by UA to HK-2 cells. Part II Effect of urea on HK-2 cells apoptosis and expression of fibrotic correlation factors and intervention of PGE1Objective:The study was designed to investigate the effect of urea on renal proximal tubular cells apoptosis and expression of fibrotic correlation factors as well as the protective effect of PGE1 against Urea-induced apoptosis and expression of fibrotic correlation factors in renal proximal tubular cellsMethods:1. HK-2 cells were used as the subject;2.The toxic effect of urea on HK-2 cells was determined by MTT assay,NAG activity and Annexin V-FTC/PI fluorescence; 3.Western Blot and Real-Time PCR were employed to investigate the expression of TGF-β1, CTGF and FN. In the blocking study, PGE1 was used as antagonists respectively.Results:HK-2 Cells were exposed to urea (100,200,400 or 800umol/L) respectively for 24h,48h,72h. Urea could induce a significant dose and time-dependent loss of cell viability. Have been treated with different concentrations Urea for 24h, NAG concentration and apoptosis ratio were increased dose dependently (P<0.01). The presence of PGE1 significantly lowered the inhibition rate of cell activity, NAG concentration and apoptotic ratio compared with the cells which were treated with Urea only.The protein and mRNA of TGF-β1, CTGF, FN in HK-2 cells increased significantly when HK-2 cells were cultivated with Urea, when PGE1 wad added in Urea fibrotic correlation factors decrease significantly.Conclusion:1.Urea could inhibition of renal proximal tubular cells proliferation and promote apoptosis in vitro; 2.Urea could promote the expression of fibrosis relation factor (TGF-β1,CTGF,FN)in renal tubular epithelial cells; 3. PGE1 can interrupt apoptosis and the expression of fibrosis correlation factor caused by Urea to HK-2 cells.