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首页> 外文期刊>Journal of nephrology. >Pirfenidone reduces in vitro rat renal fibroblast activation and mitogenesis.
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Pirfenidone reduces in vitro rat renal fibroblast activation and mitogenesis.

机译:吡非尼酮可降低体外大鼠肾成纤维细胞的活化和有丝分裂。

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BACKGROUND: Fibroblasts have been universally recognised in tubulointerstitial injury, where their presence has been shown to be a marker of disease progression. Recently, pirfenidone (PF) has been shown to both ameliorate progressive fibrosis and reduce established scarring after ureteric obstruction (UUO) in the rat, suggesting that it is a novel anti-fibrotic agent. The objective of this study was therefore to determine if these effects include down-regulation of fibroblast function. METHODS: Cortical fibroblasts were obtained from outgrowth cultures of renal tissue isolated from kidneys 3 days after UUO and constituted 100% of cells studied. Functional studies examined the effects of 20 and 200 microg/ml PF on basal serum stimulated activity. Activation was examined by western blotting for alpha smooth muscle actin (alphaSMA) and connective tissue growth factor (CTGF). Cell proliferation, collagenase activity and collagen production were determined from kinetic studies, zymography for MMP2 and [3H] proline incorporation in collagenous proteins respectively. RESULTS: Proliferation, as measured by [3H] thymidine incorporation, was reduced in dose dependent manner by 20 and 200 microg/ml PF (p<0.05; 200 vs 0 microg/ml). Likewise, 200 microg/ml PF reduced cell population growth over 5 days of culture (p<0.05 vs 0 microg/ml). PF (200 microg/ml) decreased alphaSMA and CTGF protein expression to 66+/-13 and 37+/-26% of basal levels respectively (both p<0.05 vs 0 microg/ml). Synthesis of collagen was unaffected by PF. Maximal dose of PF produced a modest reduction in MMP2 lytic activity (p=0.05). Effects of PF were independent of cell toxicity. CONCLUSIONS: Down-regulation of renal fibroblast activation and proliferation are specific actions of PF.
机译:背景:成纤维细胞已在肾小管间质损伤中得到普遍认可,其存在已被证明是疾病进展的标志。最近,吡非尼酮(PF)已显示出既可以改善进行性纤维化,又可以减少大鼠输尿管梗阻(UUO)后建立的瘢痕形成,这表明它是一种新型的抗纤维化剂。因此,本研究的目的是确定这些影响是否包括成纤维细胞功能的下调。方法:皮质成纤维细胞从UUO后3天从肾脏分离的肾脏组织的长出培养物中获得,并构成了所研究细胞的100%。功能研究检查了20和200微克/毫升PF对基础血清刺激的活性的影响。通过蛋白质印迹检查α平滑肌肌动蛋白(alphaSMA)和结缔组织生长因子(CTGF)的激活。分别从动力学研究,MMP2的酶谱分析和[3H]脯氨酸掺入胶原蛋白中确定了细胞增殖,胶原酶活性和胶原蛋白的产生。结果:通过[3H]胸苷掺入量测量的增殖以剂量依赖性方式降低了20和200 microg / ml PF(p <0.05; 200 vs 0 microg / ml)。同样,200微克/毫升PF降低了培养5天的细胞群生长(p <0.05对0微克/毫升)。 PF(200 microg / ml)分别将alphaSMA和CTGF蛋白表达降低至基础水平的66 +/- 13和37 +/- 26%(p <0.05 vs 0 microg / ml)。胶原蛋白的合成不受PF影响。 PF的最大剂量导致MMP2裂解活性适度降低(p = 0.05)。 PF的作用与细胞毒性无关。结论:肾成纤维细胞激活和增殖的下调是PF的特定作用。

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