类叶牡丹3种皂苷元对L929细胞炎症模型中HDAC表达的影响

Effects of Three Sapogenins from Cauorlhyllum Robustum Maxim on HDAC Expression in L929 Cell Inflammation Model

目的:研究类叶牡丹3种皂苷元抑制HDAC3/8进而抗类风湿性关节炎的作用。方法:建立TNF-α诱导的小鼠成纤维L929细胞炎症模型,通过CCK-8方法检测3种皂苷元的最佳给药浓度和TNF-α的最佳造模浓度,以IL-6浓度为指标,运用ELISA法验证炎症模型是否建立成功以及各成分的抗炎能力。采用蛋白免疫印迹法(Western Blot)检测L929细胞中HDAC3、HDAC8蛋白表达的变化。结果:通过CCK-8法得到TNF-α的最佳浓度为10 ng/mL,常春藤皂苷元选用浓度为500μmol/L和250μmol/L,齐墩果酸选用浓度为50μmol/L和25μmol/L,刺囊酸选用浓度为25μmol/L和12.5μmol/L。TNF-α诱导后L929细胞模型组中的IL-6含量明显升高(P<0.01),不同浓度给药组对IL-6的含量呈现出不同的抑制效果(P<0.05,P<0.01)。3种皂苷元成分会明显降低TNF-α诱导的L929细胞炎症模型的HDAC3、HDAC8蛋白表达水平(P<0.05,P<0.01)。结论:在TNF-α诱导的L929细胞炎症模型中,类叶牡丹3种皂苷元呈现出良好的抗炎效果,其作用机制可能与降低HDAC过表达有关。

Objective:To investigate the inhibitory effects of three sapogenins from Cauorlhyllum Robustum Maxim on HDAC3/8 and their anti-rheumatoid arthritis(RA)activity.Methods:A TNF-α-induced inflammation model in L929 fibroblast cells was established in mice.The optimal drug concentration of the screened components and the optimal modeling concentration of TNF-α were determined using the CCK-8 method.The inflammation model was verified by ELISA using IL-6 concentration as an indicator,and the anti-inflammatory ability of the screened components was evaluated.Changes in HDAC3 and HDAC8 protein expression in L929 cells were detected using Western blot.Results:The optimal concentration of TNF-αwas determined to be 10 ng/mL by the CCK-8 method.Hederagenin was used at concentrations of 500μmol/L and 250μmol/L,oleanolic acid at concentrations of 50μmol/L and 25μmol/L,and Echinocystic acid at concentrations of 25μmol/L and 12.5μmol/L.The IL-6 content in the L929 cell model group induced by TNF-α was significantly increased(P<0.01),and the different concentrations of drug groups showed different inhibitory effects on IL-6(P<0.05,P<0.01).The three sapogenins components significantly reduced the protein expression levels of HDAC3 and HDAC8 in the TNF-α-induced L929 cell inflammation model(P<0.05,P<0.01).Conclusion:In the TNF-α-induced L929 cell inflammation model,the three sapogenins from Cauorlhyllum Robustum Maxim showed good anti-inflammatory effects by reducing HDAC overexpression.