EGCG对硫酸吲哚酚诱导的脂肪细胞氧化应激和炎症的影响

Protective Effects of Epigallocatechin Gallate on Indoxyl Sulfate-induced Oxidative Stress and Inflammation in Adipocytes

目的:观察硫酸吲哚酚(IS)对3T3-L1脂肪细胞氧化应激和炎症的影响及EGCG干预的效应。方法:以3T3-L1脂肪细胞为研究对象,分为对照组(control)、IS组(100μmol/L、IS250μmol/L、IS500μmol/L)、IS+EGCG组(IS250μmol/L+EGCG5μmol/L、IS250μmol/L+EGCG10μmol/L、IS250μmol/L+EGCG20μmol/L)。干预24 h后,CCK-8法检测细胞活力,生化试剂盒检测细胞内活性氧(ROS)含量及NADPH氧化酶活性,ELISA方法检测细胞培养上清液中TNF-α、IL-6及MCP-1含量。结果:不同试剂干预24 h后,与对照组相比,IS100、250、500μmol/L和EGCG5、10、20μmol/L对细胞活力无影响;与对照组相比,IS250、500μmol/L组细胞内ROS水平明显增加,细胞培养上清液中TNF-α、IL-6及MCP-1明显增加,差异有统计学意义(P<0.05);加入NADPH氧化酶抑制剂apocynin后,IS250μmol/L组细胞内ROS水平明显降低,细胞培养上清液中TNF-α、IL-6及MCP-1含量明显下降,差异有统计学意义(P<0.05)。与IS250μmol/L组相比,加入EGCG10、20μmo/L干预后,细胞内ROS水平明显下降、NADPH氧化酶活性明显降低,细胞培养上清液中TNF-α、IL-6及MCP-1含量明显下降(P<0.05)。结论:EGCG可改善IS诱导的3T3-L1脂肪细胞氧化应激和炎症,其作用与其抑制NADPH氧化酶有关。

Objective:To observe the protective effect of epigallocatechin gallate(EGCG)on indoxyl sulfate(IS)-induced oxidative stress and inflammation in 3 T3-L1 adipocytes and discuss the mechanisms.Methods:3 T3-L1 adipocytes served as objects,which were cultured in various media,including IS(100μmol/L,250μmol/L,500μmol/L),IS+EGCG(IS250μmol/L+EGCG5μmol/L,IS250μmol/L+EGCG10μmol/L,IS250μmol/L+EGCG20μmol/L)for 24 h.The cell viability was determined by cell counting kit-8(CCK-8).The production of reactive oxygen species(ROS)and the activity of NADPH oxidase were determined by biochemical kits.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and monocyte chemoattractant protein-1(MCP-1)were determined by enzyme-linked immunosorbent assay(ELISA).Results:After treatment for 24 h,there was no decrease in cell viability in different IS groups and EGCG groups compared with the control group.The levels of ROS and adipocytokines(TNF-α,IL-6,MCP-1)in the IS250 and 500μmol/L groups significantly increased(P<0.05).Compared with the IS250μmol/L group,the production of ROS and the levels of TNF-α,IL-6 and MCP-1 significantly reduced after adding EGCG10,20μmol/L and the NADPH oxidase inhibitor apocynin 100μmol/L(P<0.05).Compared with the IS250μmol/L group,the activity of NADPH oxidase significantly decreased after adding EGCG10,20μmol/L.Conclusion:EGCG inhibited IS-induced oxidative stress and inflammation in 3 T3-L1 adipocytes by the suppression of the activity of NADPH oxidase.