Expression And Clinical Significance Of Beclin1,LC3,Atg9a MRNA And Oxidative Stress Markers In Rheumatoid Arthritis

objective: Our study first prepare to detect the expression of autophagy related genes Beclin1,LC3,Atg9 a mRNA from peripheral blood mononuclear cells(PBMCs) separeted from RA patients and healthy normal people peripheral blood;Collecting the serum of RA patients, healthy normal people, other autoimmune diseases patients including osteoarthritis(OA) 、 ankylosing spondylitis(AS)、gout to detect the oxidative stress indicators;Gathering the synovial fluid of RA、to detect the oxidative stress indicators levels,and analyse the correlations between autophagy related genes and oxidative stress indicators.Aiming at discussing the role of autophagy and oxidative stress in RA and the relationship among three, in order to find an effective drug treatment target,new diagnostic marker and monitoring indicator.Methods: Choosing the 54 RA patients, 44 healthy normal people meeting diagnostic standard as research objects. Acquisition the RA patients and healthy normal people peripheral blood on an empty stomach,using differential density gradient centrifugation to sepatate PBMCs then to extract the total RNA, adopting the reverse transciption PCR(RT-PCR) and real- time fluorescent quantitave PCR to detect the relative expression of Beclin1,LC3,Atg9 a mRNA. And choosing 54 RA patients,44 healthy people,24 other autoimmune diseases patients’ serum,18 RA and other autoimmune diseases patients’ joint fluid separately by utilizing colorimetry to detect the objects’ oxidative stress indicators levels,including malondialdehyde(MDA), nitric oxide(NO)and superoxide dismutase(SOD).And anlyzing the correlation beweent the thirds and Beclin1,LC3,Atg9 a mRNA.Results: The real-time PCR results : 1. The relative expression of LC3 mRNA in RA group significantly incresed compared with healthy controls(P<0.05);But Beclin1 and Atg9 a have no statistical differences between the two groups(P>0.05). 2.Compared with the prior-treatment group in RA,the Beclin1,LC3 mRNA expression of the post-treatment group significantly reduced.(P<0.05).however,the Atg9 a mRNA relative expression declined slightly after treatment and have no statistical difference.(P>0.05). 3. LC3 mRNA in RA disease activity rise and relativeexpression show difference during the forth activity periods.The higher the activity, the expression of LC3 mRNA is higher(P>0.05).While Beclin1,Atg9 a mRNA in disease activity and disease remission have no obvious differences(P>0.05).Oxidative stress test results: 1. In serum, RA group’ s levels of MDA,NO significantly increased compared with healthy controls,the levels of SOD significantly dereased(P<0.05);2.In joint synovial fluid,the levels of MDA,NO all elevated in RA group and other autoimmune disease group,but there are no statistical difference between the two.SOD levels all decrese in the two groups,but also no obvious difference(P>0.05). 3. Compared with the prior-treatment group in RA,the levels of MDA,NO is significantly reduced in the post-treatment group(P<0.05),and the levels of SOD is increased after treatment(P<0.05). 4.SOD in RA disease activity decline obviously and show difference during the forth activity periods.The higher the activity, the levels of MDA is lower(P<0.05). 5. The levels of NO are associated with autophagy related genes(r=0.389,0.394;P<0.05).Conclusion: Both of the states of increased autophagy and oxidative stress are exited in RA patients. And autophagy has relationship with reactive oxygen.Autophagy may be used as therapeutic targets to improve the condition of patients with RA,the oxidative stress index may be regarded as the new index to monitor therapeutic effect.