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目的:观察大黄酸和大黄素对人肾小管上皮细胞(HK-2)增殖及对人转化生长因子-β1(TGF-β1)基因启动子活性的作用。方法:应用MTT法观察不同浓度大黄酸和大黄素(1μg/ml、10μg/ml、50μg/ml和100μg/ml)对人肾小管上皮细胞增殖的作用;将人TGF-β1基因启动子片段与氯霉素乙酰基转移酶(CAT)报告基因组成重组体ph TGF 2.14,采用脂质体法转染至人肾小管上皮细胞中,分别加入不同浓度的大黄酸和大黄素(1μg/ml、10μg/ml和50μg/ml)进行干预,用ELISA方法检测报告基因CAT的活性。结果:大黄酸和大黄素各浓度组对人肾小管上皮细胞均具有明显的抑制HK-2细胞增殖的作用(P<0.01),且呈剂量依从性;大黄酸和大黄素浓度为1μg/ml和10μg/ml时对重组体ph TGF 2.14的表达活性无影响,当增大其浓度为50μg/ml时对重组体ph TGF 2.14的表达活性有抑制作用,其抑制率分别为38.0%和36.0%,与对照组比较均有显著差异(P<0.01)。结论:大黄酸和大黄素能够抑制人肾小管上皮细胞增殖,当浓度为50μg/ml时对人TGF-β1基因启动子活性具有一定的抑制作用。
Objective: To observe the effects of rhein and emodin on the proliferation of human renal tubular epithelial cells (HK-2) and the promoter activity of human transforming growth factor-β1 (TGF-β1) gene. Methods: The effects of different concentrations of rhein and emodin (1μg / ml, 10μg / ml, 50μg / ml and 100μg / ml) on the proliferation of human renal tubular epithelial cells were observed by MTT assay. Chloramphenicol acetyltransferase (CAT) reporter gene recombined ph TGF 2.14, was transfected into human renal tubular epithelial cells by liposome, were added different concentrations of rhein and emodin (1μg / ml, 10μg / ml and 50μg / ml) were used to detect the CAT activity of the reporter gene by ELISA. RESULTS: Both rhein and emodin concentrations significantly inhibited the proliferation of HK-2 cells in human renal tubular epithelial cells (P <0.01), and were dose-dependent. The concentrations of rhein and emodin were 1 μg / ml And 10μg / ml had no effect on the expression activity of recombinant ph TGF 2.14. When its concentration was 50μg / ml, its inhibitory rate was 38.0% and 36.0%, respectively. , Compared with the control group were significantly different (P <0.01). Conclusion: Rhein and emodin can inhibit the proliferation of human renal tubular epithelial cells. When the concentration is 50μg / ml, the activity of human TGF-β1 gene promoter may be inhibited.