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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 β(TGF β)和纤维连接蛋白(FN)的影响及其可能机制。 方法 :①分别以 10 -12 、10 -11、10 -10 、10 -9、10 -8mmol/L的hPTH1 3 4刺激大鼠系膜细胞6、12、2 4、48h后 ,ELISA方法测定上清中TGF β和FN的浓度 ;②分别以 10 -12 ～ 10 -8mol/L的hPTH1 3 4刺激大鼠系膜细胞 48h后 ,半定量RT PCR方法检测细胞TGF βmRNA的表达 ;③以 10 -8mol/L的hPTH1 3 4分别刺激大鼠系膜细胞 6～ 48h ,采用半定量RT PCR方法检测细胞TGF βmRNA的表达。④分别以 10 -12 ～ 10 -8mol/L的hPTH1 3 4和 10ng/L的抗TGF β抗体共培养大鼠系膜细胞 ,以无抗TGF β抗体为对照 ,48h后测定上清中FN的水平。⑤以 10 -8mol/L的hPTH1 3 4与 10 μg/L的抗TGF β抗体共同刺激大鼠系膜细胞 ,分别于 6～ 48h测定上清中FN的浓度 ,以无抗TGF β抗体为对照。 结果 :①ELISA结果显示 ,hPTH1 3 4促进大鼠系膜细胞合成与分泌TGF β和FN ,且具有浓度依赖和时间依赖性特点 ,当PTH浓度为 10 -8mol/L时 ,其促分泌作用达高峰 (P <0 0 1) ;②半定量RT PCR方法结果显示 ,hPTH1 3 4促进大鼠系膜细胞TGF βmRNA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 0 5 ) ;?
Objective: To investigate the effects of PTH on the synthesis and secretion of transforming growth factor β (TGFβ) and fibronectin (FN) in rat mesangial cells and its possible mechanism. Methods: ① The rat mesangial cells were stimulated with 10, 12, 10 -11, 10 -10, 10 -9, 10 -8 mmol / L hPTH1 3 4 for 6,12,2 4,48 h, respectively, The concentrations of TGFβ and FN in the serum were detected by MTT assay. ② The mesangial cells were stimulated with 10-12 ~ 10 -8 mol / L hPTH1 3 4 for 48 h respectively. The expression of TGF β mRNA was detected by semiquantitative RT PCR. The mesangial cells were stimulated with 8mol / L hPTH1 34 for 6-48 h, respectively. The expression of TGFβ mRNA was detected by semi-quantitative RT-PCR. ④ The rat mesangial cells were co-cultured with 10 -12 ~ 10 -8 mol / L hPTH1 34 and 10 ng / L anti-TGF β antibody respectively. After 48 h, the FN Level. ⑤ The mesangial cells were stimulated with 10 -8 mol / L hPTH1 3 4 and 10 μg / L anti-TGF β antibody respectively. The concentration of FN in the supernatant was measured at 6 ~ 48 h. The anti-TGF β antibody was used as control . Results: ① ELISA results showed that hPTH1 3 4 promoted the synthesis and secretion of TGF β and FN in rat mesangial cells in a concentration-dependent and time-dependent manner. When the concentration of PTH was 10 -8 mol / L, the secretion of TGF β and FN reached the peak (P <0.01). ② The results of semi-quantitative RT-PCR showed that hPTH1 3 4 promoted the expression of TGF β mRNA in rat mesangial cells in a concentration-dependent and time-dependent manner (P <0 0 5);?