盐酸小檗碱对大鼠肾小管上皮细胞缺血再灌注损伤的保护作用 张日东 .doc

盐酸小檗碱对肾小管上皮细胞缺血再灌注( （南京医科大学附属南京第一医院内分泌科，南京 210006） [摘要 目的：观察盐酸小檗碱（BBR）对缺血再灌注大鼠肾小管上皮细胞(NRK-52E)活性的影响。方法：应用化学性缺氧模拟剂氯化钴（CoCl2）在NRK-52E细胞建立化学性缺氧再给氧损伤模型以不同浓度（1×10-6mol/L、1×10-5mol/L、1×10-4mol/L）的BBR处理NRK-52E细胞。应用四甲基偶氮唑法（MTT）检测BBR对各组细胞活性的影响；检测各组细胞上清液中的丙二醛(MDA)、超氧化物歧化酶(SOD)含量；Hoechst33258染色观察各组凋亡细胞的形态学变化；Annexin-FITC/PI染色流式细胞仪检测各组细胞凋亡率。结果MTT结果显示1×10-4mol/L BBR对细胞毒性作用明显，1×10-6 mol/L、1×10-5mol/LBBR提高缺再组细胞存活率，尤以1×10-5mol/L为高；Hoechst33258显示正常组基本无凋亡细胞，模型组出现核大染的凋亡细胞，而BBR(1×10-5mol/L)预处理组凋亡细胞明显减少；与正常对照组相比，缺细胞上清液中MDA含量明显升高(P0.05)，SOD活力显著降低(P0.05)。而用BBR预处理后组的MDA显著低于用药前(P0.05)，SOD则高于组(P0.05)，上述作用在BBR浓度为1×10-6mol/L1×10-5mol/L范围内呈浓度依赖性。流式细胞仪结果显示1×10-5mol/L、1×10-6mol/LBBR预处理后细胞凋亡率分别为11.1%、32.2%与用药前56.2%比较有显著性差异(P0.05)。结论：BBR对缺血再灌注肾小管上皮细胞具有保护作用，尤以1×10-5mol/L作用最为明显。 关键词 盐酸小檗碱细胞凋亡肾小管上皮细胞缺血再灌注The studys of berberine on ischemia-reperfusion renal tubular epithelial cell injury ZHANG Ri-dong,BAI Rui-miao,Wei Jing( (Department of Endocrinology, Nanjing First Hospital affiliated to Nanjing Medical University, Nanjing 210006, China) Abstract Objective: To observe the effect of the berberine (BBR) on ischemia-reperfusion renal tubular epithelial cells (NRK-52E). Methods: Using chemical hypoxia cobalt chloride (CoCl2) to stimulate NRK-52E cells to develop a chemical model of hypoxia-reoxygenation injury, with different concentrations (1×10-6mol/L,1×10-5mol/L,1×10-4mol/L) of the BBR dealing with NRK-52E cells. Using tetrazolium method (MTT) to test the activity of the cells; detecting the malondialdehyde (MDA) and superoxide dismutase (SOD) content of each group; Hoechst33258 staining was used to obseve the morphological changes of apoptotic cells of each group; Annexin-FITC/PI staining was applied to detect the rate of apoptosis cell in each group. Results: MTT results showed that 1×10-4mol/L BBR was toxic to the cells, but 1×10-6mol/L and 1×10-5mol/L BBR could increase the ischemia-reperfusion group cell survival, especially in 1×10-5mol/L for the high; Hoechst33258 showed almost no apoptotic ce