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  

Protection Effect of Berberine Ischemia-Reperfusion for Renal Tubular Epithelial Cell of Rats

作　　者：张日东[1] 白瑞苗[1] 魏敬[1] 

机构地区：[1]南京医科大学附属南京第一医院内分泌科,南京210006

出　　处：《中国实验方剂学杂志》2011年第4期165-168,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(30271527);江苏省自然科学基金项目(BK2002142)

摘　　要：目的:观察盐酸小檗碱(berberine,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,模拟在体的缺血再灌注模型。以不同浓度(1×10-6,1×10-5,1×10-4 mol.L-1)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)含量、超氧化物歧化酶(SOD)活性;Hoechst33258染色观察各组凋亡细胞的形态学变化;Annexin V-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1×10-4 mol.L-1 BBR对细胞毒性作用明显,而1×10-6,1×10-5 mol.L-1BBR明显提高缺氧再给氧组细胞存活率,尤以1×10-5 mol.L-1为高;Hoechst33258显示正常组基本无凋亡细胞,模型组出现核大深染的凋亡细胞,而BBR(1×10-5 mol.L-1)预处理组凋亡细胞明显减少;与正常对照组相比,缺氧再给氧组细胞上清液中MDA含量明显升高(P<0.05),SOD活力显著降低(P<0.05)。而用BBR预处理后缺氧再给氧组的MDA显著低于用药前(P<0.05),SOD则高于缺氧再给氧组(P<0.05),上述作用在BBR浓度为1×10-6～1×10-5 mol.L-1呈浓度依赖性。流式细胞仪结果显示1×10-5,1×10-6 mol.L-1 BBR预处理后缺氧再给氧各组细胞凋亡率分别为11.1%,32.2%与用药前56.2%相比较有显著性差异(P<0.05)。结论:BBR对缺血再灌注的肾小管上皮细胞具有保护作用,尤以1×10-5 mol.L-1作用最为明显。Objective：To observe the effect of the berberine（BBR）on ischemia-reperfusion for renal tubu-lar epithelial cells（NRK-52E）of rat.Method： Using chemical hypoxia cobalt chloride（CoCl2） to stimulate NRK-52E cells to develop a chemical model of hypoxia-reoxygenation injury.Then,the cells were treated with different concentrations（1 × 10-6,1 × 10-5,1 × 10-4 mol.L-1）of BBR.Methylthiazolyldiphenyl-tetrazolium bromide（MTT） was used to test the activity of the cells.Malondialdehyde（MDA）and superoxide dismutase（SOD） content of each group were determined;Hoechst33258 staining was used to detect the morphological changes of apoptotic cells of each group;Annexin V-FITC /PI staining was applied to detect the rate of apoptosis cell in each group.Result： MTT results showed that 1 × 10-4 mol.L-1 BBR was toxic to the cells,but 1 × 10-6 mol.L-1and 1 × 10-5 mol.L-1BBR could increase the ischemia-reperfusion group cell survival,especially in 1 × 10-5 mol.L-1 for the high;Ho-echst33258 showed almost no apoptotic cells in the normal group,in the model group there were mostly nuclear stai-ning of apoptotic cells,while BBR（1 × 10-5 mol.L-1） pretreatment significantly reduced apoptosis cells;With the normal control groups,the MDA of ischemia-reperfusion group content was significantly higher（P 0.05）,but the SOD was significantly lower（P 0.05）.The pretreatment with BBR group,MDA was significantly lower than that of before treatment（P 0.05）,SOD was higher than the ischemia-reperfusion group（P 0.05）,the role of BBR depends on the concentration between 1 × 10-6 mol.L-1 and 1 × 10-5 mol.L-1.The Flow Cytometry showed that the apoptosis rate of 1 × 10-5,1 × 10-6 mol.L-1 BBR group and ischemia-reperfusion group were 11.1%,32.2% and 56.2%,and the differences were significantly（P 0.05）.Conclusion： The pretreatment with BBR on the ischemi-a-reperfusion cell has protective effects on renal tubular epithelial cells,especially the concentration of 1 × 10-5 mol.L-1 was most effectiv

关 键 词：盐酸小檗碱 细胞凋亡 肾小管上皮细胞 缺血再灌注 

分 类 号：R285.5[医药卫生—中药学]