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目的 :通过研究低氧对肾小管上皮细胞的作用 ,探讨肾间质纤维化的机制及抗氧化剂lazaroid的保护作用。 方法 :建立化学性低氧环境 ,应用台盼蓝染色作活细胞计数 ,并应用流式细胞技术检测肾小管上皮细胞凋亡。 结果 :应用无糖、无血清培养液并加用antimycinA建立化学性低氧环境。化学性低氧作用肾小管上皮细胞 12h后流式细胞仪可检测到明显亚二倍体subG1峰 ,提示出现明显的细胞凋亡 ,与对照组比较差异显著 (其百分比分别为 6 46± 0 32和 2 6 9± 0 5 1,P <0 0 1) ,而同时应用抗氧化剂lazaroid可减少低氧引起的细胞凋亡 ,与对照组比较差异不显著 (其百分比分别为 2 5 6± 0 5 2和 2 6 9± 0 5 1,P >0 0 5 )。同时台盼蓝拒染计数显示相似的结果 ,存活细胞数在化学性低氧作用 12h后逐渐减少。 结论 :低氧对肾小管上皮细胞具有直接的致细胞凋亡作用 ,这可能在肾间质纤维化中起重要作用 ,而抗氧化治疗可防止小管细胞的损伤
OBJECTIVE: To investigate the mechanism of hypoxia on renal tubular epithelial cells and explore the mechanism of renal interstitial fibrosis and the protective effect of lazaroid. Methods: To establish a chemical hypoxic environment, trypan blue staining for viable cell count, and the use of flow cytometry detection of renal tubular epithelial cell apoptosis. Results: The application of sugar-free, serum-free medium plus antimycinA to establish a chemical hypoxic environment. Chemical hypoxic effects of tubular epithelial cells 12h after flow cytometry subdiploid sub-peak was detected obvious subtle, suggesting significant apoptosis, compared with the control group significant difference (the percentage was 6 46 ± 0 32 And 269 ± 0 5 1, P <0.01), while the application of lazaroid at the same time could reduce the hypoxia-induced apoptosis, with no significant difference compared with the control group (the percentages were 256 ± 0 5 2 and 269 ± 0 5 1, P> 0 0 5). At the same time, trypan blue staining showed similar results. The number of viable cells decreased gradually after 12h of chemical hypoxia. CONCLUSIONS: Hypoxia has a direct apoptotic effect on renal tubular epithelial cells, which may play an important role in renal interstitial fibrosis, whereas anti-oxidant therapy prevents tubule cell injury