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目的研究过氧化物酶体增殖物激活受体γ(PPARγ)激动剂阻断转化生长因子(TGF)β1致肾间质纤维化作用的机制,探讨其抗肾间质纤维化的潜在作用。方法体外培养大鼠肾成纤维细胞株(NRK/49F),观察PPARγ配体15d-PGJ2及其激动剂曲格列酮和齐格列酮对TGFβ1诱导的纤维连接蛋白(FN)mRNA表达的影响。利用Western印迹技术观察PPARγ激动剂对TGFβ1诱导的FN和Smad蛋白表达的影响。结果(1)与1ng/ml TGFβ1组比较,5ng/ml TGFβ1组FN mRNA表达量增加了3·6倍(P<0·01),5ng/ml TGFβ1刺激24h时较刺激前增加了2·4倍(P<0·01),TGFβ1诱导FNmRNA表达呈一定范围内的剂量(0~5ng/ml)和时间(0~24h)依赖效应。(2)与5ng/mlTGFβ1组比较,10μmol15d-PGJ2、曲格列酮和齐格列酮预处理组FN mRNA表达量分别降低37·3%、41·5%和22·7%,FN蛋白表达量分别降低20·6%、38·1%和28·6%。(3)5ng/ml TGFβ1以时间(0~2h)依赖方式诱导p-Smad2/3蛋白表达量增加,作用1h时达到高峰;5ng/ml TGFβ1组p-Smad2/3蛋白表达量较对照组和2ng/ml TGFβ1组分别增加3·42倍和0·97倍。(4)15d-PGJ2、曲格列酮和齐格列酮预处理组p-Smad2/3蛋白表达量与5ng/ml TGFβ1组比较分别降低61·2%、53·0%和59·5%,3种药物干预组之间p-Smad2/3蛋白表达量比较差异无统计学意义,各组Smad2和Smad3蛋白表达量无显著变化。结论PPARγ激动剂可以抑制TGFβ1诱导的肾间质成纤维细胞细胞外基质合成,其机制可能与阻断TGF-β1/Smad信号途径有关,提示PPARγ激动剂具有抗肾间质纤维化的潜在作用,可能成为延缓终末期肾功能衰竭的治疗新手段之一。
Objective To investigate the mechanism of peroxisome proliferator - activated receptor γ (PPARγ) agonist blocking the TGF - β1 - induced interstitial fibrosis and to explore its potential role in anti - renal interstitial fibrosis. Methods Rat renal fibroblast cell line (NRK / 49F) was cultured in vitro and the effect of PPARγ ligand 15d-PGJ2 and its agonists troglitazone and ziglitazone on TGFβ1-induced fibronectin (FN) mRNA expression . The effect of PPARγ agonist on TGFβ1-induced FN and Smad protein expression was observed by Western blotting. Results (1) Compared with 1ng / ml TGFβ1 group, the expression of FN mRNA in 5ng / ml TGFβ1 group increased 3.6 times (P <0.01) (P <0.01). TGFβ1 induced a dose-dependent (0-5ng / ml) and time (0-24h) dependent effect on FN mRNA expression. (2) Compared with 5ng / ml TGFβ1 group, the expression of FN mRNA decreased by 37.3%, 41.5% and 22.7% in 10μmol 15d-PGJ2, troglitazone and ziglitazone preconditioning groups, respectively. FN protein expression Decreased by 20.6%, 38.1% and 28.6% respectively. (3) The expression of p-Smad2 / 3 in 5ng / ml TGFβ1 induced by time (0 ~ 2h) -dependent increased the peak at 1h, and the expression of p-Smad2 / 3 in 5ng / 2ng / ml TGFβ1 group increased by 3.42 times and 0.97 times. (4) Compared with 5ng / ml TGFβ1 group, the expression of p-Smad2 / 3 in 15d-PGJ2, troglitazone and ziglitazone pretreatment groups decreased by 61.2%, 53.0% and 59.5% There was no significant difference in the expression of p-Smad2 / 3 among the three drug-intervention groups, but the expression of Smad2 and Smad3 in each group did not change significantly. Conclusion PPARγ agonist can inhibit the TGFβ1-induced extracellular matrix synthesis of renal interstitial fibroblasts, and its mechanism may be related to blocking the TGF-β1 / Smad signaling pathway, suggesting that PPARγ agonists have a potential anti-renal interstitial fibrosis, It may become one of the new treatments for delaying end-stage renal failure.