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目的:通过观察甲状旁腺素(PTH)对体外培养的人肾小管上皮细胞转分化和转化生长因子-β1(TGF-β1)表达的影响,以探讨PTH在肾间质纤维化发病机制中的作用。方法:体外培养人近端肾小管上皮细胞(HK-2),分别用不同浓度的PTH(10-12、10-11、10-10、10-9、10-8mol/L)作用于HK-2细胞48h,以及10-8mol/LPTH作用于HK-2细胞不同时间(12、24、48、72h)。应用RT-PCR法检测HK-2细胞内α-SMA、TGF-β1mRNA表达水平,免疫细胞化学法检测HK-2细胞表达α-SMA阳性细胞百分数,Western blot法检测细胞内α-SMA的蛋白表达水平。结果:(1)RT-PCR结果表明,PTH呈剂量和时间依赖性地促进HK-2细胞α-SMA、TGF-β1mR-NA的表达(P<0.01),且两者之间呈正相关(P<0.05)。(2)免疫细胞化学结果表明,PTH可增加表达α-SMA阳性的HK-2细胞百分数,且呈剂量和时间依赖性(P<0.05)。(3)Western blot结果表明,PTH可增加HK-2细胞α-SMA的蛋白表达量,也呈剂量和时间依赖性(P<0.01)。结论:PTH可通过促进肾小管上皮细胞转分化以及TGF-β1的表达促进肾小管间质纤维化。
OBJECTIVE: To investigate the effect of PTH on the transdifferentiation of human renal tubular epithelial cells and the expression of transforming growth factor-β1 (TGF-β1) in vitro to explore the role of PTH in the pathogenesis of renal interstitial fibrosis effect. Methods: Human proximal tubular epithelial cells (HK-2) were cultured in vitro and treated with different concentrations of PTH (10-12,10-11,10-10,10-9,10-8 mol / L) 2 cells 48h, and 10-8mol / LPTH HK-2 cells at different times (12,24,48,72 h). The expression of α-SMA and TGF-β1 mRNA in HK-2 cells was detected by RT-PCR. The percentage of α-SMA positive cells in HK-2 cells was detected by immunocytochemistry. The protein expression of α-SMA was detected by Western blot Level. Results: (1) RT-PCR results showed that PTH promoted the expression of α-SMA and TGF-β1mR-NA in HK-2 cells in a dose- and time- <0.05). (2) Immunocytochemistry results showed that PTH increased the percentage of HK-2 cells expressing α-SMA, in a dose-and time-dependent manner (P <0.05). (3) Western blot results showed that PTH increased the expression of α-SMA in HK-2 cells in a dose-and time-dependent manner (P <0.01). Conclusion: PTH can promote tubulointerstitial fibrosis by promoting renal tubular epithelial cell transdifferentiation and TGF-β1 expression.