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目的:探讨缺氧性肾小管细胞损伤机制及丹参的保护作用。方法:将人近曲肾小管上皮细胞随机分为对照组、模型组和丹参组,每组均设立6个时间点包括缺氧4、12、24 h和缺氧24 h后复氧4、12、24 h进行观察,其中丹参组设立0.10%、0.20%、0.30%、0.40%0、.50%5个浓度(生药浓度);采用倒置相差显微镜观察各组细胞生长状况及形态学改变,MTT法活细胞计数检测细胞成活率,放射免疫分析法检测培养细胞上清液TNF-α含量。结果:与对照组比较,模型组随缺氧时间延长,细胞数量逐渐减少,并伴形态改变,24 h达高峰,复氧后细胞数量增加,形态改变不明显;浓度0.10%丹参组细胞损伤改善最明显;TNF-α在缺氧/复氧各时间点均降低。结论:TNF-α参与了缺氧性肾小管细胞损伤病理过程;丹参对缺氧性肾小管细胞损伤具有保护作用。
Objective: To investigate the mechanism of hypoxic tubular cell injury and the protective effect of Salvia Miltiorrhiza. METHODS: Human proximal renal tubular epithelial cells were randomly divided into control group, model group and Salvia miltiorrhiza group. Each group was established with 6 time points including hypoxia at 4, 12, 24 h and hypoxia 24 h after reoxygenation. Observations were made at 24 h, in which Danshen group was established with 0.1%, 0.20%, 0.30%, 0.40%0, and .50% of 5 concentrations (raw drug concentration); the growth status and morphological changes of each group were observed using an inverted phase contrast microscope. MTT The viable cell count was used to test the viability of the cells. Radioimmunoassay was used to measure the supernatant of cultured cells. RESULTS: Compared with the control group, the model group gradually decreased with the hypoxia time, and the number of cells gradually decreased with morphological changes and reached a peak at 24 hours. After reoxygenation, the number of cells increased and the morphology was not obvious; the concentration of 0.10% Salvia miltiorrhiza cell damage was improved. The most obvious; TNF-α decreased at all time points of hypoxia/reoxygenation. Conclusion: TNF-α is involved in the pathological process of hypoxic tubular cell injury; Salvia miltiorrhiza has protective effect on hypoxic tubular cell injury.