黄芪当归合剂及他汀类药物对缺氧复氧人肾小管上皮细胞的保护作用

The protective role of Astragalus-Angelica mixture and Atorvastatin on renal tubular epithelial cells with hypoxia reoxygenation

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.