论文部分内容阅读
目的 探讨肾上腺髓质素 (AM)在人肾小管上皮细胞系 (HK 2 )对转化生长因子 β1(TGF β1)生物学效应的影响。观察系膜增生性肾小球肾炎患者血浆AM水平与肾小管间质病变的关系。方法 体外细胞培养实验分为 4组 (1)TGF β1组 ;(2 )AM组 ;(3)AM +TGF β1组 ;(4 )对照组。以3H TdR掺入法测定细胞增殖 ;细胞总胶原的合成和分泌以3H 脯氨酸掺入量及培养液内3H 羟脯氨酸放射性活性来判断 ;ELISA法检测培养液中的纤连蛋白 (FN)含量 ;FN、IV型胶原和TIMP 1mRNA的表达采用RT PCR法 ;2 5例系膜增生性肾小球肾炎患者血浆AM水平测定采用放射免疫法。并设对照组。结果 (1)AM对TGF β1的生长抑制作用无明显影响 ;(2 )AM呈浓度依赖性抑制TGF β1刺激的胶原合成和分泌 ,与TGF β1组相比 ,AM(10 -8mol/L)分别抑制了 8% (P >0 .0 5 )和 30 % (P <0 .0 5 )的胶原合成和分泌 ,AM(10 -7mol/L)则分别抑制了 5 7% (P <0 .0 5 )和 6 4% (P <0 .0 1) ;并且AM明显抑制TGF β1刺激的FN分泌 ,AM +TGF β1组FN分泌 (2 0ng/ μl± 5ng/ μl)较TGF β1组 (2 8ng/ μl± 6ng/ μl)降低了 30 % (P <0 .0 5 ) ;AM显著抑制TGF β1刺激的IV型胶原、FN及TIMP 1mRNA的表达 ;(3)肾小管间质轻度病变组和重度病变组患者血浆AM水平较对照组分别增
Objective To investigate the effect of adrenomedullin (AM) on the biological effects of transforming growth factor β1 (TGFβ1) in human renal tubular epithelial cell line (HK 2). To investigate the relationship between plasma AM levels and tubulointerstitial lesions in patients with mesangial proliferative glomerulonephritis. Methods The in vitro cell culture experiments were divided into four groups: (1) TGFβ1 group; (2) AM group; (3) AM + TGFβ1 group; (4) control group. Cell proliferation was measured by 3H TdR incorporation assay. 3H-proline incorporation and 3H-hydroxyproline radioactivity in cells were determined by the synthesis and secretion of total collagen. ELISA was used to detect the expression of fibronectin FN) content; FN, type IV collagen and TIMP 1 mRNA expression using RT PCR method; 25 cases of mesangial proliferative glomerulonephritis in patients with plasma AM levels by radioimmunoassay. And set the control group. Results (1) AM had no significant effect on the growth inhibition of TGFβ1. (2) AM inhibited the collagen synthesis and secretion stimulated by TGFβ1 in a concentration-dependent manner. Compared with TGFβ1 group, AM (10-8mol / L) Inhibited collagen synthesis and secretion by 8% (P> 0.05) and 30% (P <0. 05), while AM (10 -7 mol / L) inhibited 5 7% 5 and 6 4% respectively (P <0.01). And AM significantly inhibited FN secretion stimulated by TGFβ1. FN secretion in AM + TGFβ1 group (20ng / μl ± 5ng / μl) (P <0.05); AM significantly inhibited the expression of type IV collagen, FN and TIMP-1 mRNA stimulated by TGF-β1; (3) mild tubulointerstitial lesions and Patients with severe disease plasma levels of AM were increased compared with the control group