雷藤舒调控滑膜关节相关长链非编码RNA2在胶原诱导性关节炎小鼠中的作用机制研究OA

Objective This study investigates the effect of wide-interacting nude-associated matrix-associated region 2(WAKMAR2)on synovial inflammation and bone destruction in collagen-induced arthritis(CIA)mice,as well as the mechanism of intervention by 5R-5-hydroxytriptolide(LLDT-8,T8).Methods The mice were randomly divided into the normal group(WT group),the CIA model group(CIA group),the overexpression empty vector group(CIA-NC group),the overexpression lentivirus group(CIA-LV group),the T8 administration group(CIA+T8 group),the T8 administration group after overexpression empty vector(CIA-NC+T8 group),and the T8 administration group after overexpression lentivirus(CIA-LV+T8 groups),8 in each group.Except for the normal group,the CIA models of mice were constructed in the other groups,and WAKMAR2-overexpressing lentivirus was injected into the knee joint cavity of mice in CIA-LV group and CIA-LV+T8 group.The virus was injected continuously for 3 weeks,once a week.The CIA group only received intragastric administration of hydroxypropyl methylcellulose(HPMC).The CIA+T8 group,the CIA-NC+T8 group and the CIA-LV+T8 group all received T8 gavage.The gavage began on the 28th day and lasted for 4 weeks.In vivo imaging was used to determine viral transfection efficiency.The expression of tumor necrosis factor-α(TNF-α),interleukin-1(IL-1),and interleukin-6(IL-6)in mouse blood was detected by enzyme-linked immunosorbent assay(ELISA)assay.The degree of joint bone destruction in mouse paws was observed using Micro-CT.Real-time quantitative PCR was used to measure the expression of WAKMAR2 and microRNA-4478(miR-4478).Western blot was used to detect the expression of E2F1 and p53 proteins.Results ①In vivo imaging demonstrated high expression of WAKMAR2 after lentiviral injection in the knee joints of mice.②Compared to the CIA group and CIA-NC group,the CIA-LV group showed significant improvement in general condition and arthritis inflammation index,with decreased expression of TNF-α,IL-1,and IL-6 in serum,increased mRNA levels of WAKMAR2,decreased mRNA levels of miR-4478,and elevated protein expression of E2F transcription factor 1(E2F1)and p53.③Compared to CIA+T8 group and CIA-NC+T8 group,CIA-LV+T8 group showed significant improvement in general condition and arthritis inflammation index,with decreased expression of TNF-α,IL-1,and IL-6 in serum,increased mRNA levels of WAKMAR2,decreased mRNA levels of miR-4478,and elevated protein expression of E2F1 and p53.Conclusions Overexpression of WAKMAR2 can improve the synovial inflammation microenvironment in CIA mice and may serve as a potential target for the treatment of rheumatoid arthritis(RA).T8 can inhibit inflammation levels in CIA mice by regulating the WAKMAR2/miR-4478/E2F1/p53 axis.These findings provide new directions for RA treatment strategies and establish a theoretical foundation for future clinical applications.