丹酚酸A、B组分配伍对HK-2细胞C C L2、CXCL10及氧化应激反应的影响

摘要

Objective To observe the effect of compatibility of salvianolic acid A and B on the expression of monocyte chemoattractant protein-1 (CCL2),CXC chemokine ligand 10 (CXCL10)and oxidative stress reaction in renal fibrosis model cells HK-2,and to explore the protective effect of salvianolic acid A and B on renal fibrosis.Methods HK-2 cells were cultured and then were divided into the normal group,model group,salvianolic acid A group,salvianolic acid B group,and salvianolic acid A +B group.The renal fibrosis cell models were all prepared in the model group,salvianolic acid A group,salvianolic acid B group,and salvianolic acid A +B group.The salvianolic acid A group was added with 20μmol/L salvianolic acid A,the salvianolic acid B group with 20 μmol/L salvianolic acid B,the salvianolic acid A +B group with 10 μmol/L salvianolic acid A,B,and the model group and normal group were not interfered.Each group was cultured for 24,48,and 72 h.The levels of CCL2 and CXCL10 in the supernatant was determined by enzyme linked im-munosorbent assay,the levels of SOD and GSH in supernatant were detected by water WST-1 method and DTNB method, the intracellular MDA levels were detected by TBA.Results The CCL2,CCL10,MDA and GSH levels in the model group were all higher than those in the normal group,and the SOD level was lower than that in the normal group (all P<0.05).The levels of CCL2,CCL10,GSH,and MDA in the supernatant of salvianolic acid B group,salvianolic acid A group and salvianolic acid A+B group were all lower than those of the model group,and the SOD level was higher than that of the model group (all P<0.05).The levels of CCL2,CCL10,MDA and GSH at each time point in the supernatant of salvianolic acid A+B group were lower than those of the salvianolic acid A group and salvianolic acid B group.The levels of SOD were higher than those of the salvianolic acid A and salvianolic acid B group (all P<0.05 ).Conclusion The compatibility of salvianolic acid A and B can inhibit the expression of inflammatory chemokines CCL2 and CXCL10 in HK-2 cells,the effect of anti-inflammation and anti-oxidation is better than that of single component application,and the combi-nation of the two components has synergistic protective effect on renal fibrosis.%目的：观察丹酚酸A、B组分配伍对HK-2肾小管上皮细胞肾纤维化模型炎症趋化因子2（CCL2）、CXC趋化因子配体10（CXCL10）及氧化应激反应的影响，探讨丹酚酸A、B组分配伍对肾纤维化的治疗作用机制。方法培养HK-2细胞，分为正常组、模型组、丹酚酸A组、丹酚酸B组、丹酚酸A＋B组。模型组、丹酚酸A组、丹酚酸B组、丹酚酸A＋B组均制备肾纤维化细胞模型。丹酚酸A组加入丹酚酸A 20μmol／L，丹酚酸B组加入丹酚酸B 20μmol／L，丹酚酸A＋B组加入丹酚酸A、B各10μmol／L，模型组及正常组不干预。各组均分别培养24、48、72 h。应用酶联免疫吸附法检测上清液CCL2、CXCL10水平，应用水溶性四唑盐法、二硫代二硝基苯甲酸法检测上清液SOD、GSH水平，应用硫代巴比妥酸法检测细胞内MDA水平。结果模型组各时点上清液CCL2、CXCL10、GSH、MDA水平均高于正常组，SOD水平均低于正常组（P均＜0．05）。丹酚酸A组、丹酚酸B组、丹酚酸A＋B组各时点上清液CCL2、CXCL10、GSH、MDA水平均低于模型组，SOD水平均高于模型组（P均＜0．05）。丹酚酸A＋B组各时点上清液CCL2、CXCL10、GSH、MDA水平均低于丹酚酸A组及丹酚酸B组，SOD水平均高于丹酚酸A组及丹酚酸B组（P均＜0．05）。结论丹酚酸A、B组分配伍可抑制HK-2细胞炎症趋化因子CCL2、CXCL10的表达，在抗炎及抗氧化方面的效果优于单一组分应用，二者配伍应用对肾纤维化具有协同治疗作用。