摘要
目的 研究肝细胞生长因子 (HGF)对 IL- 1α诱导的肾小管上皮细胞肌纤维母细胞转分化 (TEMT)及细胞分泌功能的影响。方法 在体外培养的正常大鼠肾小管上皮细胞 (NRK5 2 E)中加入 IL- 1α2 0 ng/ml诱导 ,并加入大 (16 0 0 ng/ml)、中 (12 0 0 ng/m l)、小 (80 0 ng/ml)剂量 HGF阻断 ,流式细胞仪和免疫组化法检测 α-平滑肌肌动蛋白的表达 ;倒置相差显微镜、扫描电镜观察细胞形态变化 ;EL ISA法测定培养细胞上清液分泌的粘连蛋白(FN)含量。结果 IL- 1α刺激后细胞转变为类似成纤维细胞形态 ;α- SMA阳性细胞百分数及 α- SMA表达的平均荧光强度明显增加 (P<0 .0 5 ) ;培养上清液中 FN含量增加 (P<0 .0 5 )。 HGF可剂量依赖性地抑制 IL- 1α诱导NRK5 2 E细胞形态学改变及 α- SMA的表达 (P<0 .0 5 ) ,不同程度地抑制 IL- 1α的促 FN分泌作用 (P<0 .0 5 )。单独加入不同剂量 HGF对肾小管细胞无影响。结论 HGF可抑制 IL- 1α诱导的 TEMT作用 ,减少 FN的分泌 。
Objective To observe the effects of hepatocyte growth factor (HGF) on tubular epithelial-myofibroblast transdifferentiation(TEMT)triggered by IL-1α and the fibronectin secretion of TEMT. Methods The normal rat kidney tubular epithelial cell line (NRK52E) was cultured for six days on plastic or collagen type I-coated plates in the presence or absence of HGF or IL-1α. The morphology of transdifferentiation tubular cells was observed by scanning electron microscopy (SEM) and phase-contrast microscopy. The number of α-SMA + cells, the percentage of α-SMA + cells and the mean channel fluorescence (MCF) were assessed by immunohistochemistry and flowcytometry. The level of fibronectin in supernatant was measured by ELISA. Results The NRK52E cells triggered by IL-1α became fibroblast-like morphologyically, and strong α-SMA immunostaining of those cells was seen. The level of FN in the culture supernatant, the percentage of α-SMA + cells and the MCF of cells triggered by IL-1α were obviously higher than those of blank control group (P<0 05). In the groups with IL-1α and different doses of HGF, the transdifferentiation of NRK52E cells was inhibited. With the increase of HGF dose, the percentage of α-SMA + cells and the level of FN showed a tendency to decrease. There was no significant difference between the groups treated with only HGF at different dose levels and the blank control group (P>0 05). Conclusion IL-1α can induce tubular epithelial cell to transdifferentiate to myofibroblast and increase the secretion of FN. These results suggest that TEMT may play an important role in the pathogenesis of renal fibrosis. HGF could block the transdifferentiation of tubular epithelial cell and inhibit the secretion of FN. These would provide a novel therapeutic strategy for the treatment of renal interstitial fibrosis and end stage renal disease.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2005年第1期9-12,共4页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金 (批准号 3 0 170 43 7
3 960 0 71)
国家教委留学回国人员启动基金 ( C2 990 0 5 )资助
