王枣子总黄酮影响佐剂性关节炎大鼠病理分子机制研究.docVIP

王枣子总黄酮影响佐剂性关节炎大鼠病理分子机制研究 　　[摘要] ?n题组前期研究发现宿州地方特色药材王枣子中总黄酮（total flavonoids of Isodon amethystoides，TFIA）对佐剂性关节炎（adjuvant arthritis，AA）具有一定的治疗作用，并且这种治疗作用可能是通过对miR152表达的上调实现的，该文进一步研究TFIA对AA大鼠病理机制影响的分子机制。采用完全弗氏佐剂制备AA大鼠，TFIA灌胃治疗，采用Realtime qPCR检测TFIA灌胃治疗对AA大鼠关节滑膜成纤维样滑膜细胞（fibroblast like synoviocytes，FLS）中miR152、甲基化酶DNMT1及甲基化结合蛋白MeCP2构成的负调控环路的影响，对miR152下游经典Wnt信号通路的影响，对AA大鼠病理基因fibronectin表达的影响。TFIA灌胃治疗显著抑制DNMT1表达，逆转了AA大鼠病理中存在的由miR152，DNMT1和MeCP2构成的负调控环路，向治疗组FLS转染miR152 inhibitors后，DNMT1表达显著恢复。TFIA灌胃治疗显著上调SFRP4表达，抑制经典Wnt信号通路关键基因βcatenin，Cmyc和ccnd1表达，显著抑制AA病理基因fibronectin表达，向治疗组FLS转染miR152 inhibitors后，逆转了TFIA灌胃治疗对SFRP4，βcatenin，Cmyc，ccnd1和fibronectin表达的影响。TFIA可能通过miR152表达的上调抑制DNMT1表达，上调SFRP4表达，抑制经典Wnt信号关节基因βcatenin，Cmyc，ccnd1表达，抑制RA基因fibronectin表达。 　　[关键词] 王枣子总黄酮； 佐剂性性关节炎； miR152； 成纤维样滑膜细胞； 经典Wnt信号通路 　　Molecular mechanism of total flavonoids in Isodon amethystoides 　　on adjuvant arthritis in rats 　　MIAO Chenggui1*， SHI Weijing1， WEI Wei1， QIN Meisong1， CHEN Hao1， ZHANG Bing2 　　（1. Food and Drug College， Anhui Science and Technology University， Fengyang 233100， China； 　　2. The First Affiliated Hospital， Anhui Medical University， Heifei 230032， China） 　　[Abstract] Our preliminary study showed that the total flavonoids in Isodon amethystoides（TFIA）， a local medicinal herb in Suzhou， had a certain therapeutic effect on adjuvant arthritis， and this therapeutic effect may be achieved through the upregulation of miR152 expression. In this paper， the molecular mechanism of TFIA on the pathogenesis of adjuvant arthritis（AA） rats was further studied. AA rats were prepared with complete Freund′s adjuvant， and then treated with TFIA by intragastric administration. Realtime qPCR was used to detect the effects of TFIA on the negative regulatory loop of miR152， methylase DNMT1 and methylCpG binding protein MeCP2 in fibroblast like synoviocytes（FLS） of AA rats， as well as the effects of TFIA on the classic Wnt signaling pathway and the expression of fibronectin gene in AA rats. In