Modified Cheng’s Juanbi Decoction Effect On Synovial Fibroblasts Of Adjuvant Arthritis Rats Beclin1、LC3B Expression

Objective:Explore Modified Cheng ’s Juanbi decoction(Modified Cheng ’s Juanbi decoction,CJBD)for Rheumatoid arthritis(Rheumatoid Rrthritis,RA)mechanism,observe CJBD high,medium and low doses of Adjuvant arthritis(Adjuvant arthritis,AA)rat’s general status,body weight,bilateral digit volume,polyarthritis index(arthritis index,AI)changes,as well as to the cytokines in serum Interleukin 4(Interleukin 4,IL-4),Interleukin 17(Interleukin 17,IL-17)in synovial fibroblasts Beclin1,LC3 B mRNA and protein expression related indicators of the level,research CJBD synovial fibroblasts of AA rats,the impact of autophagy for RA CJBD treatment curative effect to provide reliable experimental basis.to achieve the efficacy of CJBD treatment RA to provide reliable experimental basis.Methods:Rats were randomly divided into normal group,model group,CJBD low,medium and high dose groups,MTX group.In addition to the normal group,0.15 ml of Freund’s adjuvant complete(CFA)was injected into the right hind toe area of rats of other groups.On the 1st day after inflammation,adopt the way of "combined disease" progresses on the model of AA enhancement of cold dampness environment stimulate the formation of AA rat model.After the model was successfully replicated,the rats in the low,medium and high dose groups of CJBD were given the corresponding doses of CJBD solution;MTX group was given the corresponding dose MTX suspension mixed liquid lavage;the model group and the normal group were given the corresponding doses of distilled water;Each group continued to administer for 28 days.Assess CJBD experimental rats weight before and after therapy,bilateral digit volume,the influence of the AI;Enzyme-linked immunosorbent(Enzyme linked immunoadsorption,ELISA)to detect the rat serum interleukin 4(Interleukin 4,IL-4),interleukin 17(Interleukin 17,IL-17);Quantitative Real-time PCR detection in the synovial tissue of rats Beclin1,LC3 B mRNA expression level;Protein imprinting method(Western blotting,WB)detection in the synovial tissue of rats Beclin1,LC3 B protein expression level.Results:1.General condition,body weight is found that compared with normal group,AA rats depression treatment,food and drink less water,inflammatory,side and secondary side digits red swelling significantly,restricted movement,accompanied by rough hair,nasal congestion,and so on and so forth,showed a trend of progression and slow growth in weight;CJBD compared with model group,high,medium and low dose group and MTX group after treatment of AA rats general symptoms abated,weight growth improved(P<0.05);2.Bilateral digit volume is found that compared with normal group,model group,the MTX group,CJBD high,medium and low dose group of dosing the toes on both sides before the volume significantly increased(P<0.05);CJBD compared with model group,high,medium and low dose group and MTX group after treatment of AA rats toes volume decreases,and on both sides and CJBD dose groups,in high dose group is superior to the CJBD CJBD dose group is superior to the CJBD in low dose group(P<0.05);3.AI is found that,compared with model group,CJBD high,medium and low dose group and MTX group AA rats after treatment of AI score down(P<0.05),which the AI ratings downgrade CJBD high dose group is better than that of MTX group(P<0.05),with CJBD MTX group there was no statistically significant difference in the dose group(P>0.05),CJBD dose group and CJBD there are differences between the low dose group(P<0.05);4.The levels of IL-4 and IL-17 testing found that compared with normal group,model group rats serum IL-4 expression level decreased obviously,the expression of IL-17 level significantly increased(P<0.05);Compared with model group,CJBD low,medium and high dose group of rats IL-4 the expression level of progressive increase,IL-17 progressive lower(P<0.05);5.Beclin1,LC3 B mRNA and protein expression level detection showed that rats compared with normal group,model group rats synovial tissues Beclin1,LC3 B mRNA and protein expression levels were significantly decreased(P<0.05);Compared with model group,Beclin1,LC3 B mRNA and protein expression in CJBD high,medium and low dose group and MTX group obviously increased(P<0.05);Compared with CJBD medium and low dose group,high dose group of rats Beclin1,LC3 B gene and protein expression level was higher(P<0.05),and low dose group,expression level is higher than CJBD dose group(P<0.05).Conclusion:1.CJBD can obviously improve AA rat’s general symptoms and slow growth,weight of ratings downgrade AI,reduce the digit joint swelling;2.The mechanism of CJBD for Rheumatoid arthritis may be related to enhance the expression of IL-4,inhibit expression of IL-17,regulating cytokine imbalance;3.CJBD may also the mechanism of treatment of RA and improve Beclin1,LC3 B mRNA and protein expression level,improve the level of cell autophagy;4.CJBD dose and the therapeutic effect of AA rats in a dose dependent,with the increase of CJBD dose,to improve the symptoms of AA rats,reduce the destruction of the joints,relieve the illness has more advantages and effects.