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机构地区:[1]南方医科大学附属花都医院肾内科,510800 [2]南方医院广东省肾脏研究所
出 处:《临床肾脏病杂志》2012年第12期562-566,共5页Journal Of Clinical Nephrology
基 金:广东省科技计划(2010B060900009)
摘 要:目的研究黄芪当归合剂及阿托伐他汀对缺氧/复氧人肾小管上皮细胞损伤的影响。方法选用人肾小管上皮细胞建立缺氧/复氧损伤模型,设正常对照组、单纯缺氧复氧组、黄芪当归合剂组、阿托伐他汀组、黄芪当归合剂和阿托伐他汀联合用药组。检测活性氧的产生量。酶联免疫吸附法检测细胞间黏附分子-1、单核细胞趋化蛋白-1的表达,逆转录聚合酶链式反应检测细胞间黏附分子-1mRNA、单核细胞趋化蛋白-1mRNA表达,免疫印迹法检测核因子-kB。结果缺氧复氧组活性氧高于对照组,细胞间黏附分子-1、单核细胞趋化蛋白-1及各自mRNA表达上调,黄芪当归合剂组、阿托伐他汀组均降低细胞内活性氧水平,抑制炎性因子表达,黄芪当归合剂和阿托伐他汀联合用药组效果更加明显。结论黄芪当归合剂、阿托伐他汀对肾小管上皮细胞缺氧复氧性损伤有保护作用,联合用药效果显著,可能与通过抑制核因子-kB炎症通路减少细胞间黏附分子-1、单核细胞趋化蛋白-1表达从而降低氧化应激水平有关。Objective To investigate the protective role and the mechanism of Astragalus-An- gelica mixture(AAM) and Atorvastatin(ATO) on renal tubular epithelial cetls(HK-2) with hypoxia reoxygenation(H/R). Methods The cultured HK-2 were divided into normal control group, H/R group, 100 μg/ml Astragalus-Angelica mixture + H/R group, 10 μmol/L Atorvastatin + H/R group, 100μg/ml Astragalus-Angelica mixture combine 10μmol/L Atorvastatin + H/R group. Reac- tive oxygen species(ROS) production in the cell was evaluated by kinetic measurement of dichlorofluo- roscein (DCF) fluorescence produced by oxidation of an oxidant sensitive dye 2,7-dichlorefluorescin (DCFH) using Victor 1420 multilabel counter. The expression of intercellular adhesinmolecule-1 (ICAM-1) and monocyte chemotactic protein-1 (MCP-1) were measured by ELISA and respectively mRNA were measured by reverse transcription PCR(RT-PCR) and the transcription factors nuclear factor-kB(NF-kB) use analysis by Western blotting. Results The levels of ROS and NF-kB in the H/ R group increased remarkably comparing with the normal control group and the expression of ICAM- 1 ,MCP-1 and respectively mRNA also increased. Treatment with Astragalus-Angelica mixture and Atorvastatin significantly reduced the oxidative stress level and curb the expression of adhesion mole- cules. Drug combination group had the best result. Conclusions Astragalus-Angelica mixture and Atorvastatin play a role in the protection against renal tubular epithelial cells with hypoxia reoxygen- ation and drug combination have more significant effect. It might reduce this hypoxia reoxygenation in- jury by blocking NF-kB inflammatory pathway decreasing the production of ICAM-1, MCP-1 and inhi- bition of cell oxidation stress.
分 类 号:R322.61[医药卫生—人体解剖和组织胚胎学]
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