王枣子总黄酮影响佐剂性关节炎大鼠病理的分子机制研究  

Molecular mechanism of total flavonoids in Isodon amethystoides on adjuvant arthritis in rats

作　　者：缪成贵[1] 时维静[1] 魏伟 秦梅颂[1] 陈浩 张兵[2] 

机构地区：[1]安徽科技学院食品药品学院,安徽凤阳233100 [2]安徽医科大学第一附属医院,安徽合肥230032

出　　处：《中国中药杂志》2017年第17期3411-3416,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81302783);安徽科技学院科研项目(ZRC2014473)

摘　　要：课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miR-152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Real-time 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miR-152、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miR-152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miR-152,DNMT1和MeCP2构成的负调控环路,向治疗组FLS转染miR-152 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β-catenin,C-myc和ccnd1表达,显著抑制AA病理基因fibronectin表达,向治疗组FLS转染miR-152 inhibitors后,逆转了TFIA灌胃治疗对SFRP4,β-catenin,C-myc,ccnd1和fibronectin表达的影响。TFIA可能通过miR-152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β-catenin,C-myc,ccnd1表达,抑制RA基因fibronectin表达。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 up-regulation of miR-152 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. Real-time q PCR was used to detect the effects of TFIA on the negative regulatory loop of miR-152,methylase DNMT1 and methyl-Cp G 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. Intragastric administration of TFIA significantly inhibited the expression of DNMT1 and reversed the negative regulatory loop composed of miR-152,DNMT1 and MeCP2 in the pathology of AA rats. After transfection of miR-152 inhibitors into the FLS in treatment group,DNMT1 expression was significantly restored. TFIA significantly up-regulated the expression of SFRP4 and inhibited the expression of β-catenin,C-myc and ccnd1,the key genes of canonical Wnt signaling pathway.TFIA also significantly inhibited the expression of fibronectin,an AA gene. The effect of TFIA on the expression of SFRP4,β-catenin,C-myc,ccnd1 and fibronectin was reversed after transfection with miR-152 inhibitors in the treatment group FLS. TFIA may inhibit the DNMT1 expression,up-regulate the SFRP4 expression,inhibit the expression of classical Wnt signaling genes β-catenin,C-myc,and ccnd1 as well as the RA gene fibronectin expression through the up-regulation of miR-152 expression.

关 键 词：王枣子总黄酮 佐剂性性关节炎 miR-152 成纤维样滑膜细胞 经典Wnt信号通路 

分 类 号：R285.5[医药卫生—中药学]