中国药理学通报2017，Vol.33Issue(6)：781-787,7.DOI:10.3969/j.issn.1001-1978.2017.06.009
橙皮素衍生物-XIV调控PTCH1基因甲基化对佐剂性关节炎大鼠FLS炎症的影响
Effect of Hesperidin derivative-XIVregulating methylation of PTCH1 gene on FLSinflammation of adjuvant arthritis rats
摘要
Abstract
Aim To explore the effect of 5-(4-Chlorophenethyl) imino-7-O-acetyl-4-chlorophenethylamine hesperetin(HDND-XIV) on fibroblast-like synoviocytes(FLS) inflammation in adjuvant arthritis(AA) rats and the underlying molecular mechanism.Methods AA rats were induced by Freund′s complete adjuvant(FCA,0.01 ml·kg-1),after 24 d,the rats were killed and the synovial tissue was used to extract primary cells;MTT was used to detect the half maximal inhibitory concentration(IC50) of HDND-XIV;Enzyme linked immunosorbent assay(ELISA) was used to detect the level of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β) in cell culture.Lipo-fectamine2000 transfection kit was used to display the overexpression of MeCP2 in AA FLS.Western blot was used to detect the expression of MeCP2,PTCH1,Gli1 and Shh.Pyrosequencing was used to detect the methylation level of the PTCH1 gene quantificationally.Results The IC50 of HDND-XIV on FLS was 161 μmol·L-1 through MTT.The most significant anti-inflammatory concentration was 10 μmol·L-1 through ELISA.In AA FLS,the expression of PTCH1 was increased and MeCP2,Gli1,Shh were reduced in response to HDND-XIV.According to pyrosequencing,the methylation level of the PTCH1 gene was significantly decreased in response to HDND-XIV.Overexpression of MeCP2 in AA FLS followed by treating with HDND-14,the expression of PTCH1 was reduced,the expression of Gli1,Shh was increased and the level of IL-6, IL-1β, TNF-α rose.Conclusion HDND-XIV has prominent anti-inflammatory activity in FLS,the mechanism of which may be associated with inhibiting the expression of MeCP2 to reduce the methylation status of PTCH1 gene.关键词
HDND-XIV/SD大鼠/佐剂性关节炎/甲基化/Hedgehog/成纤维滑膜细胞/过表达/炎症Key words
HDND-XIV/Sprague Dawley rats/adjuvant arthritis/methylation/Hedgehog/fibroblast-like synoviocytes/overexpression/inflammation分类
医药卫生引用本文复制引用
孙正浩,刘彦会,刘骏达,徐丹丹,李小枫,张义龙,孟晓明,黄成,李俊..橙皮素衍生物-XIV调控PTCH1基因甲基化对佐剂性关节炎大鼠FLS炎症的影响[J].中国药理学通报,2017,33(6):781-787,7.基金项目
国家自然科学基金资助项目(No 81273526,81473268) （No 81273526,81473268）
安徽省教育厅重点基金项目(No KJ2016A364,KJ2016A365) （No KJ2016A364,KJ2016A365）
安徽省自然科学基金面上项目(No 21408085MKL31) （No 21408085MKL31）
