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目的 探讨内皮素 1(ET 1)及ET 1受体A拮抗剂 (ETaRA)对人肾间质成纤维细胞(hRIF)的作用。方法 在体外培养的hRIF中进行如下试验 :(1)MTT比色法检测细胞增殖情况。 (2 )逆转录多聚酶链反应法 (RT PCR)观察Ⅰ型胶原 (ColⅠ )、转化生长因子 β(TGF β)、基质金属蛋白酶 1(MMP 1)、金属蛋白酶组织抑制物 1,2 (TIMP 1,TIMP 2 )mRNA表达的变化。结果 (1)ET 1(10 -11～10 -7mol/L ,2 4h)可以促进hRIF增殖 ,10 -7mol/L增殖最显著 (P <0 0 5 ) ,呈剂量相关 ;10 -7mol/L刺激 8h ,hRIF即明显增殖 ,2 4h达高峰 (P <0 0 1)。 (2 )ET 1(10 -11～ 10 -7mol/L ,16h)可上调hRIF的ColⅠ、TGF β、MMP 1、TIMP 1、TIMP 2mRNA表达 ,10 -7mol/L作用最显著 (P <0 0 5 ) ,呈剂量相关。10 -7mol/L刺激hRIFs时 ,ColImRNA在 8h表达显著增加 (P <0 0 5 ) ,2 4h达高峰 ;TGF βmRNA在 8h表达显著增加 (P <0 0 5 )且达高峰 ;TIMP 1及TIMP 2mRNA在 16h表达显著增加 (P <0 0 5 ) ,2 4h达高峰 ;MMP 1mRNA在 16h表达显著增加 (P <0 0 5 ) ,且达高峰。 (3)上述作用可特异地被ETaRA阻断。结论 ET 1可刺激hRIF增殖 ,并上调ColⅠ、TGF β、MMP 1、TIMP 1、TIMP 2mRNA的表达 ,ETaRA可抑制上述反应。ET 1可能在疾病状态下参与肾间质
Objective To investigate the effect of endothelin 1 (ET 1) and ET 1 receptor antagonist (ETaRA) on human renal interstitial fibroblasts (hRIF). Methods The following experiments were conducted in hRIF cultured in vitro: (1) Cell proliferation was detected by MTT assay. (2) Reverse transcriptase-polymerase chain reaction (RT PCR) was used to observe the expression of collagen type Ⅰ, transforming growth factor β (TGFβ), matrix metalloproteinase 1 (MMP 1), tissue inhibitor of metalloproteinase 1,2 , TIMP 2) mRNA expression changes. Results (1) ET 1 (10 -11 ~ 10 -7 mol / L, 24 h) could promote the proliferation of hRIF, the most obvious proliferation was 10 -7 mol / L (P <0.05) After stimulated for 8h, hRIF significantly proliferated and peaked at 24 h (P <0.01). (2) The expression of ColⅠ, TGFβ, MMP-1, TIMP-1, and TIMP-2 mRNA in hRIF cells was increased by ET 1 (10 -11-10 -7 mol / L, 5), was dose-related. At 10 -7 mol / L stimulation of hRIFs, the expression of ColImRNA increased significantly at 8h (P <0 05) and peaked at 24 hrs; the expression of TGFβmRNA increased significantly at 8h (P 0 05) 2 mRNA increased significantly at 16h (P <0 05) and peaked at 24 h. The expression of MMP-1 mRNA increased significantly at 16h (P <0 05), reaching its peak. (3) The above effects can be specifically blocked by ETaRA. Conclusion ET 1 can stimulate the proliferation of hRIF and up-regulate the expressions of ColⅠ, TGFβ, MMP-1, TIMP-1 and TIMP-2 mRNA. ETaRA can inhibit the above reaction. ET 1 may be involved in renal interstitium in disease states