title: miR-375 regulates the canonical Wnt pathway through FZD8 silencing in arthritis synovial fibroblasts.
abstract: Whether the rheumatoid arthritis (RA) pathogenesis is regulated by microRNA (miRNA) is not entirely clear. In this study, we found that miR-375 was down-regulated significantly in fibroblast-like synoviocytes (FLS) in adjuvant-induced arthritis (AIA) rat model compared with control. Because the web-based software TargetScan and PicTar predict Frizzled 8 (FZD8) as the target of miR-375, we investigated whether up-regulated miR-375 plays a role in the activation of the canonical Wnt signaling by targeting the FZD8. Furthermore, the purpose of the present experiments was also to determine the role of miR-375 in the pathogenesis of AIA rat model and to ascertain the effects of FZD8 in this process. Real time qPCR, Western blotting, ELISA and ChIP assay were used to assess the inhibited role of miR-375 in the pathogenesis of AIA rat model and the canonical Wnt signaling. RNA interference was also used to detect the role of knockdown of dephosphorylated β-catenin. Luciferase reporter gene and related methods were performed to determine the FZD8 as the target of miR-375. The increased miR-375 inhibited the pathogenesis of AIA rat model as indicated by decreases in the several disease markers, such as MMP3 and fibronectin. Interestingly, miR-375 also inhibited the canonical Wnt signaling, and the stabilized form of β-catenin blocked the miR-375 effects. FZD8 was identified as the target of miR-375 in AIA rat model by the firefly luciferase reporter gene. In summary, our results demonstrate that miR-375 regulates the pathogenesis of AIA rat model through the canonical Wnt signaling pathway. This discovery may provide new targets for therapeutic intervention to benefit RA patients.
authors.affiliation: School of Food and Drug, Anhui Key Laboratory of Poultry Epidemic Prevention and Surveillance, Anhui Science and Technology University, Fengyang 233100, China; School of Pharmacy, Anhui Medical University, Meishan Road, Hefei, Anhui 230032, China.
authors.forename: Cheng-gui
authors.initials: CG
authors.lastname: Miao
authors.affiliation: School of Food and Drug, Anhui Key Laboratory of Poultry Epidemic Prevention and Surveillance, Anhui Science and Technology University, Fengyang 233100, China.
authors.forename: Wei-jing
authors.initials: WJ
authors.lastname: Shi
authors.affiliation: School of Food and Drug, Anhui Key Laboratory of Poultry Epidemic Prevention and Surveillance, Anhui Science and Technology University, Fengyang 233100, China.
authors.forename: You-yi
authors.initials: YY
authors.lastname: Xiong
authors.affiliation: School of Food and Drug, Anhui Key Laboratory of Poultry Epidemic Prevention and Surveillance, Anhui Science and Technology University, Fengyang 233100, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Yu
authors.affiliation: School of Food and Drug, Anhui Key Laboratory of Poultry Epidemic Prevention and Surveillance, Anhui Science and Technology University, Fengyang 233100, China.
authors.forename: Xiao-lin
authors.initials: XL
authors.lastname: Zhang
authors.affiliation: School of Food and Drug, Anhui Key Laboratory of Poultry Epidemic Prevention and Surveillance, Anhui Science and Technology University, Fengyang 233100, China.
authors.forename: Mei-song
authors.initials: MS
authors.lastname: Qin
authors.affiliation: School of Food and Drug, Anhui Key Laboratory of Poultry Epidemic Prevention and Surveillance, Anhui Science and Technology University, Fengyang 233100, China.
authors.forename: Chuan-lai
authors.initials: CL
authors.lastname: Du
authors.affiliation: Affiliated Hospital, Bengbu Medical College, Bengbu 233000, China.
authors.forename: Tong-wen
authors.initials: TW
authors.lastname: Song
authors.affiliation: School of Pharmacy, Anhui Medical University, Meishan Road, Hefei, Anhui 230032, China. Electronic address: lijunamu@126.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Li
mesh_terms: D000818:Animals; D001169:Arthritis, Experimental / Q000235:genetics* / Q000378:metabolism*; D001172:Arthritis, Rheumatoid / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D005347:Fibroblasts / Q000378:metabolism*; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D034622:RNA Interference; D051381:Rats; D011956:Receptors, Cell Surface / Q000235:genetics*; D013583:Synovial Membrane / Q000378:metabolism*; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000235:genetics
chemical_list: C504124:FZD8 protein, rat; C574979:MIRN375 microRNA, rat; D035683:MicroRNAs; D011956:Receptors, Cell Surface; D051176:beta Catenin
keywords: Adjuvant-induced arthritis; Canonical Wnt signaling; Frizzled 8; Rheumatoid arthritis; miR-375