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目的:探讨温莪术对转化生长因子(TGF-β1)诱导的肾小管上皮细胞转分化的作用及机制。方法:通过体外培养肾小管上皮细胞(NRK-52E)及体外药物血清技术,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,倒置显微镜观察细胞形态学改变,细胞免疫荧光法检测各组肾小管上皮细胞α-平滑肌肌动蛋白(α-SMA)mRNA表达的变化,实时荧光定量PCR法检测细胞整合素连接激酶(ILK)mRNA表达的变化,ELISA法检测细胞上清液中胶原Ⅰ(Col-Ⅰ)的分泌水平。结果:TGF-β1可诱导NRK-52E向成纤维细胞转化,出现肥大、拉长,显现梭形,胞浆内α-SMA表达明显增多(P<0.05),ILKmRNA的表达、Col-Ⅰ的分泌水平升高(P<0.05)。温莪术呈浓度依赖性抑制转分化发生,减少形态变化,下调α-SMA、ILK、Col-Ⅰ表达(P<0.05)。结论:TGF-β1可诱导转分化发生,伴有α-SMA、ILK、Col-Ⅰ等表达升高,而温莪术可抑制TGF-β1诱导的转分化,减少α-SMA、ILK、Col-Ⅰ的表达水平,并且有浓度依赖性。
Objective: To investigate the effect and mechanism of Curcuma on the transdifferentiation of renal tubular epithelial cells induced by transforming growth factor-β1 (TGF-β1). Methods: The effects of different concentrations of Rhizoma Curcumae containing different serums on the cell proliferation were detected by MTT assay, the morphological changes of the cells were observed by inverted microscope, and the expression of NRK-52E was detected by immunofluorescence The changes of α-smooth muscle actin (α-SMA) mRNA expression in renal tubular epithelial cells of each group were detected by real-time quantitative PCR. The changes of ILK mRNA expression were detected by ELISA. Ⅰ (Col-Ⅰ) secretion levels. RESULTS: TGF-β1 induced the transformation of NRK-52E to fibroblasts, and showed hypertrophy and elongation. The expression of α-SMA in cytoplasm was significantly increased (P <0.05), the expression of ILK mRNA and the secretion of Col-Ⅰ Elevated levels (P <0.05). Curcuma curcumin inhibited the transdifferentiation in a concentration-dependent manner, reduced the morphological changes and down-regulated the expression of α-SMA, ILK and Col-Ⅰ (P <0.05). CONCLUSION: TGF-β1 can induce transdifferentiation with increased expression of α-SMA, ILK and Col-Ⅰ, while Curcuma could inhibit TGF-β1-induced transdifferentiation and decrease α-SMA, ILK and Col-Ⅰ The level of expression, and concentration-dependent.