摘要
目的观察缺氧复氧后肾小管上皮细胞肿瘤坏死因子α(TNFα)的表达及胡黄连提取物对其的影响。方法建立人肾小管上皮细胞缺氧复氧损伤模型,分别用不同浓度胡黄连提取物(10μg/mL、100μg/mL、1 000μg/mL)进行预处理,采用流式细胞术检测不同组细胞内氧化应激水平,ELISA法检测细胞上清TNFα蛋白表达、RT-PCR法测定细胞TNFαmRNA表达。结果缺氧复氧后肾小管上皮细胞内氧化应激水平提高,细胞TNFα蛋白和mRNA表达上调,且随缺氧时间的延长,细胞内氧化应激水平逐渐升高,TNFα表达逐渐增强;胡黄连提取物呈剂量关系抑制细胞内氧化应激水平,同时呈剂量关系抑制TNFα表达的上调。结论缺氧复氧导致肾小管上皮细胞内氧化应激增强,进而诱导TNFα表达上调;胡黄连提取物可以通过抑制细胞内氧化应激抑制TNFα的上调。
Objective To investigate the mechanism of expression of TNFα in renal tubular epithelial cells with hypoxia-reoxygenation (H/R) and to evaluate the effects of extracts of Picrorhiza scrophlariiflora Pennell on it. Methods Cultured human renal epithelial HK2 cells were divided into five groups: normal control group, H/R group, 10 μg/mL extracts of Picrorhiza scrophlariiflora Pennell + H/R group, 100 μg/mL extracts of Picrorhiza scrophlariiflora Pennell + H/R group, and 1 000 μg/mL extracts of Picrorhiza scrophlariiflora Pennell + H/R group. Flow cytometry was used to detect the oxidative stress and expressions of TNFα protein. Real-time quantitative reverse transcfiption-polymerase chain reaction (RT-PCR) was used to detect the expression of TNFα mRNA. Result The oxidative stress in renal tubular epithelial cells was induced by H/R. TNFα protein was increased after H/R. TNFα mRNA was increased after H/R. Oxidative stress in the renal tubular epithelial cells was inhibited by extracts of Picrorhiza scrophlariiflora Pennell in a dose-dependent manner. TNFα protein was inhibited by extracts of Picrorhiza scrophlariiflora Pennell in a dose-dependent manner. TNFα mRNA was inhibited by extracts of Picrorhiza scrophlariiflora Pennell in a dose-dependent manner.
出处
《中国比较医学杂志》
CAS
2013年第10期31-35,共5页
Chinese Journal of Comparative Medicine
关键词
缺氧复氧
肾小管上皮细胞
肿瘤坏死因子
胡黄连提取物
Extracts of Picrorhiza scrophlariiflora Pennell protects renal epithelial cells against TNFα by inhibiting