柚皮苷通过抑制ERK1/2活化和DRP1表达减轻棕榈酸处理后脂肪细胞炎症

Naringin Alleviates Adipocyte Inflammation after Palmitic Acid Treatment through Inhibiting ERK1/2 Activation and DRP1 Expression

目的探讨柚皮苷(Nar)对棕榈酸(PA)处理后成熟脂肪细胞炎症反应的影响及机制。方法3T3-L1前脂肪细胞经成脂化诱导为成熟脂肪细胞后采用不同浓度(0、25、50、100、200μmol·L^(-1))PA处理细胞48 h,CCK-8检测细胞活力,ELISA检测细胞上清IL-6水平,确定后续实验最佳PA浓度。不同浓度(0、12.5、25、50、100μmol·L^(-1))Nar处理成熟脂肪细胞,CCK-8检测细胞活力,确定后续实验最佳Nar浓度。单独PA处理实验分为2组:Ctrl组、PA组;联合Nar处理实验分为4组:Ctrl组、PA组、Nar组、PA+Nar组;western blotting法检测各组p-ERK1/2和DRP1蛋白表达水平。联合线粒体裂变抑制剂Mdivi-1处理实验分为7组:Ctrl组、PA组、Nar组、Mdivi-1组、PA+Nar组、PA+Mdivi-1组、PA+Nar+Mdivi-1组,ELISA和western blotting法分别检测7组细胞上清IL-6水平和p-ERK1/2、DRP1蛋白表达水平。结果后续实验PA和Nar分别选择50和25μmol·L^(-1)。与Ctrl组比较,PA组p-ERK1/2和DRP1表达水平均升高(P<0.001)。与PA组比较,PA+Nar组、PA+Mdivi-1组、PA+Nar+Mdivi-1组IL-6水平、p-ERK1/2和DRP1表达均更低(P<0.05);与PA+Nar组比较,PA+Nar+Mdivi-1组IL-6水平差异无统计学意义(P>0.05),但p-ERK1/2和DRP1表达水平均降低(P<0.05)。结论Nar可减轻PA处理后成熟脂肪细胞的炎症反应,其机制是通过抑制ERK1/2激活和DRP1表达实现的。

Objective To investigate the effects and mechanism of narinin(Nar)on inflammatory response of mature adipocytes after palmitic acid(PA)treatment.Methods After 3T3-L1 pre-adipocytes were induced into mature adipocytes by lipogenesis,the cells were treated with different concentrations(0,25,50,100,200μmol·L^(-1))of PA for 48 h.Cell viability was detected by CCK-8,and the level of IL-6 in the culture supernatant was detected by ELISA to determine the optimal PA concentration for subsequent experiments.Mature adipocytes were treated with different concentrations(0,12.5,25,50,100μmol·L^(-1))of Nar,and cell viability was detected by CCK-8 to determine the optimal Nar concentration for subsequent experiments.PA treatment experiments were divided into 2 groups:Ctrl group and PA group;Nar treatment experiments were divided into 4 groups:Ctrl group,PA group,Nar group,and PA+Nar group;western blotting was used to detect the protein expression levels of p-ERK1/2 and DRP1 in each group.Mitochondrial fission inhibitor Mdivi-1 treatment experiments were divided into 7 groups:Ctrl group,PA group,Nar group,Mdivi-1 group,PA+Nar group,PA+Mdivi-1 group,and PA+Nar+Mdivi-1 group;ELISA and western blotting were used to detect the levels of IL-6 in the supernatant of the 7 groups,and the levels of p-ERK1/2 and DRP1 protein expression,respectively.Results In follow-up experiments PA and Nar were selected at 50 and 25μmol·L^(-1),respectively.p-ERK1/2 and DRP1 expression levels were elevated in the PA group compared with the Ctrl group(P<0.001).Compared with the PA group,IL-6 level,p-ERK1/2 and DRP1 expression were lower in the PA+Nar,PA+Mdivi-1,and PA+Nar+Mdivi-1 groups(P<0.05);compared with the PA+Nar group,the difference in IL-6 level was not statistically significant in the PA+Nar+Mdivi-1 group(P>0.05),but p-ERK1/2 and DRP1 expression levels were both reduced(P<0.05).Conclusion The results suggest that Nar can alleviate the PA-induced adipocyte inflammation by inhibiting ERK1/2 activation and DRP1 expression.