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目的:探讨温莪术对人转化生长因子β1(TGF-β1)诱导的肾小管上皮细胞转分化及细胞外基质纤维连接蛋白(FN)、胶原Ⅰ(Col-Ⅰ)的影响。方法:将体外培养的肾小管上皮细胞(NRK-52E)随机分为6组:正常血清组、TGF-β1诱导模型组、洛汀新组、温莪术高剂量组、温莪术中剂量组、温莪术低剂量组。其中温莪术及洛汀新以含药血清方式给药,运用MTT法检测不同浓度温莪术含药血清对细胞增殖的影响,确定适合的药物作用范围。应用倒置显微镜观察细胞的形态学改变,实时荧光定量PCR法检测Col-ⅠmRNA、FN mRNA的表达水平。结果:TGF-β1可诱导肾小管上皮细胞(NRK-52E)向成纤维细胞转化,细胞出现肥大、拉长,显现梭形,并且使Col-Ⅰ、FNmRNA的表达水平升高(P<0.05)。而加入不同浓度温莪术后,细胞的形态接近于正常细胞,Col-Ⅰ、FNmRNA的表达均显著下降(P<0.05),并且呈现药物浓度依赖性。结论:温莪术可抑制TGF-β1诱导肾小管上皮细胞向成纤维细胞转分化,并且出现Col-Ⅰ、FNmRNA表达的升高,并且有浓度依赖性。
Objective: To investigate the effect of Curcuma on the transdifferentiation of renal tubular epithelial cells and the expression of extracellular matrix fibronectin (FN) and collagen Ⅰ (Col-Ⅰ) induced by transforming growth factor β1 (TGF-β1). Methods: NRK-52E cells were randomly divided into 6 groups: normal serum group, TGF-β1-induced model group, Lotensin group, high-dose Curcuma group, middle-dose Curcuma group, Curcuma low-dose group. Which Curcuma and Lotensin drug-containing serum administration, the use of MTT assay of different concentrations of Curcuma containing serum on cell proliferation, determine the scope of appropriate drugs. The morphological changes of cells were observed by inverted microscope, and the expression of Col-ⅠmRNA and FN mRNA were detected by real-time fluorescence quantitative PCR. Results: TGF-β1 induced the transformation of renal tubular epithelial cells (NRK-52E) into fibroblasts. The cells showed hypertrophy, elongation, fusiform appearance and increased expression of Col-Ⅰ and FN mRNA (P <0.05) . After addition of different concentrations of Curcuma, the morphology of cells was close to that of normal cells. The expression of Col-Ⅰ and FN mRNA decreased significantly (P <0.05), and the concentration of Col-Ⅰ and FN mRNA showed drug concentration dependence. Conclusion: Curcuma could inhibit the TGF-β1-induced renal tubular epithelial cells transdifferentiation into fibroblasts, and the expression of Col-Ⅰ, FN mRNA expression increased, and in a concentration-dependent manner.