Study On The Mechanism Of MiRNAs Regulating Activation Of RASFs And Anti-inflammatory Effect Of Resveratrol

Background:Rheumatoid arthritis(RA)is a chronic,inflammatory,autoimmune disease that primarily affects the joints.As the disease progresses,the disease can eventually lead to joint destruction or even disability.Synovial fibroblasts play an important role in the the formation of joint destruction by activation of inflammation.Synovial fibroblasts in established rheumatoid arthritis usually express fixed-pattern microRNAs.This kind of non-encoded single-stranded RNA structure,which is about 22 nucleotides in length and conserved in morphology and diverse in function,can directly or indirectly bind different genes in cells to interfere with gene expression,affect protein synthesis and regulate the function of cells.MiRNA-126 has been reported to regulate the expression of PI3K-AKT signaling pathway by targeting PIK3R2 gene in a variety of tumor-related diseases,and to promote cell proliferation and reduce apoptosis.Furthermore,the natural compound resveratrol has been proved to have the function of antagonizing phosphorylated AKT expression in the PI3K/AKT signaling pathway in rheumatoid arthritis synovial fibroblasts,and has multiple functions such as anti-inflammatory,immune regulation,and anti-osteoporosis,suggesting that it may be developed into an effective drug for controlling the rheumatoid arthritis.Purpose:To investigate the effects of modify the microRNAs expression levels on the proliferation and apoptosis of rheumatoid arthritis synovial fibroblasts and the specific molecular mechanisms that produce this effect;Also,to demonstrate that resveratrol inhibits inflammatory cytokines,immune regulation and anti-inflammatory effects in the treatment of rheumatoid arthritis.Method:In this paper,we use lentiviral expression vectors to over express the level of miRNAs in RASFS,then real-time polymerase chain reaction and western blotting were used to dect the gene and the protein levels,respectively.MTT assay and flow cytometry were used to mesure the differences of cell proliferation and cell cycle,respectively.In the second part experiment of this paper,the molecular docking was designed to detect the affinity between resveratrol and TNF-α as well as IL-1.In addition,Immunomodulator activity of resveratrol was evaluated using oxidative burst assay.Result:Our data indicate that miRNA-126 overexpression in RASFs inhibits PIK3R2 expression and promotes proliferation while inhibiting apoptosis through regulating PI3K/AKT signal pathway.The molecular docking showed mechanistic inhibition of TNF-α and IL-1 by resveratrol and showed us that TNF-a is a better suited target for reseveratrol inhibition for RA treatment.Moreover,the neutrophil respiratory burst test confirmed the cytoprotective effect of resveratrol.Conclusion:The experimental results predict that the inhibition of miRNA126 expression in fibroblast-like synoviocytes can be developed as a treatment for the control of rheumatoid arthritis joint destruction.The natural compound Resveratrol,which is extracted from Polygonum cuspidatum,has the efficacy of inhibiting the progression of rheumatoid arthritis by inhibiting inflammatory factors,protecting cells,and resisting bone destruction,and may continue to be researched and developed to an effective drug in controlling rheumatoid arthritis.