摘要
目的 研究甲状旁腺激素(PTH)对大鼠系膜细胞合成与分泌转化生长因子β1(TGF-β1)的影响。方法 (1)分别以10-12、10-11、10-10、10-9、10-8mol/l的hPTH1-34刺激大鼠系膜细胞6、12、24、48h后,ELISA方法测定上清中TGF-β1的浓度;(2)分别以10-12、10-11、10-10、10-9、10-8mol/L的hPTH1-34刺激大鼠系膜细胞48h,采用半定量RT-PCR方法检测细胞TGF-β1mRNA的表达;(3)以10-8mol/L的hPTH1-34分别刺激大鼠系膜细胞6、12、24、48h,采用半定量RT-PCR方法检测细胞TGF-β1mRNA的表达。结果(1)ELISA结果显示,hPTH1-34促进大鼠系膜细胞合成与分泌TGF-β1,且具有浓度依赖和时间依赖性特点,当PTH浓度为10-8mol/L时,其促TGF-β1分泌作用达高峰(P<0.01);(2)半定量RT-PCR方法结果显示,hPTH1-34促进大鼠系膜细胞TGF-β1mRNA的表达,并具有浓度依赖和时间依赖性特点(各组与对照组间P<0.05)。结论 hPTH1-34从蛋白和基因水平显著促进系膜细胞TGF-β1的合成与分泌,且呈浓度依赖和时间依赖性特点。
Objective To identify whether PTH can regulates the synthesis and secretion of TGF-B1 in cultured mesangial cells of rats. Methods (1) Mesangial cells seeded at a density of 1 X 104 cells/well in 24-well plates were treated with medium containing various concentrations of hPTH1-34 (10-12 - 10-8mol/L) for different time (6 h, 12 h, 24 h and 48 h) ; while control cells were treated with vehicle only. Then TGF-B1 levels of the supernatants were measured by ELISA. (2) Mesangial cells seeded at a density of 3 x 105 cells/well in 6-well plates were treated with medium containing various concentrations of hPTH1-34(10-12- 10-8mol/L) .48 hours later, TGF-BlmRNA was tested by half quantitative RT-PCR. (3) The same way as before, cells were treated with 10-8mol/L hPTH1-34 for 6, 12, 24, 48 hours. Then TGF-BlmRNA was tested by half quantitative RT-PCR. Results (1)hPTH1-34 stimulated TGF-B1 synthesis in a dose- and time-dependent way with a peak at 10-8mol/L( P < 0. 01) . (2) hPTH1-34 increased the expression of TGF-B1 mRNA of cultured rat mesangial cells in a dose-and time-dependent manner ( P < 0.05) . Conclusion hPTH1-34 up-regulates the protein synthesis and mRNA expression of TGF-B1 in cultured mesangial cells of rats.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2002年第2期120-122,共3页
Chinese Journal of Nephrology