The Study Of Acupuncture On Th1/Th2 Related Cellular Molecules And Foxp3 Expression In RA Mice

Objective:By observing the improvement of arthritis symptoms in RA mice by acupuncture,the expression of Treg cells in peripheral blood and the differentiation status of Th1 and Th2 cytokines in RA mice after acupuncture were detected.To explore the effect of Treg cells on the differentiation pattern of Th1/Th2 cells after acupuncture,in order to provide a basis for acupuncture to improve the mechanism of RA arthritis.Methods:Twenty-four healthy male Wistar mice were randomly divided into a blank group,a model group,and a needle group.In the model group and the acupuncture group,the rheumatoid arthritis model was prepared by using bovine type II collagen and Freund's adjuvant,and the blank group was routinely raised.In the acupuncture group,the "Jinshu" and "Zusanli" points on both sides of the mice were treated with acupuncture.Acupuncture needles for 30 minutes every day,continuous acupuncture for 6 days for a course of treatment,a total of 3 courses of acupuncture,1 day after each treatment.The remaining groups were bound to the acupuncture group but were not treated.The body mass and the foot volume of the mice were measured before and after the modeling,before and after the treatment,ie(days 1,14,21,42),and the joint inflammation index(AI)score of the mice was observed.The knee joint tissue was taken and the morphology of the ankle joint was observed by HE staining.Detection of IL-2,IL-4,IFN-? and TNF-? in peripheral blood of mice by enzyme-linked immunosorbent assay(ELISA).The expression levels of IL-2,IFN-? and IL-4 in cells were detected by polymerase chain reaction(PCR).Flow cytometry(FCM)was used to detect the effect ofacupuncture on the expression of Foxp3 on the surface of Treg cells.Result:1.Body mass,foot volume,AI score and histopathological changes of joint synovium: compared with the blank group,the body weight of the model group was reduced(P<0.01);the volume of the foot was significantly increased(P<0.01);the AI score was also significantly improved(P<0.01);the articular cartilage injury,synovial cell invasion,and fibrous tissue proliferation were significantly increased(P<0.01),synovial cell proliferation,and macrophages were proliferated(P<0.05).Compared with the model group,the body weight of the acupuncture group increased significantly(P<0.05);the paw volume and AI score decreased(P<0.05?P<0.01);the inflammatory cell invasion decreased significantly(P<0.05),fibrous tissue proliferation was significantly reduced(P <0.01).2.The concentration of Th1 and Th2 related cytokines in peripheral blood of RA mice and the results of gene expression: Compared with the blank group,the concentrations of IL-2,TNF-? and IFN-? in the peripheral blood of the model group increased,and the concentration of IL-4 decreased.(P<0.01);IL-2 and IFN-?gene expression in cells increased(P<0.05),IL-4 gene expression decreased significantly(P<0.01);Compared with the model group,the concentrations of IL-2,TNF-? and IFN-?in the peripheral blood of the acupuncture group were lower(P<0.05),and the concentration of IL-4 was increased(P<0.05).IL-2 in the cells.The expression of IFN-? gene was decreased(P<0.05),and there was no significant difference in IL-4gene expression(P>0.05).3.The expression of Treg cell marker FoxP3 in peripheral blood of RA mice:Compared with the model group,the positive expression of CD4+CD25+Foxp3+was increased in the acupuncture group(P<0.01).Conclusions:1.Acupuncture can improve the arthritic symptoms of RA mice,increase the body mass of the mice,resolve the redness and swelling of the knee joint,reduce the AI score,and reduce the infiltration of inflammatory cells and the proliferation of fibrous tissue in the knee joint tissues of mice,and improve the damage of synovial membrane and the destruction of bone and joint.2.Acupuncture can reduce Th1 inflammatory cytokine content and gene expression,increase Th2 marker cytokine content and Treg cell marker molecule FoxP3 expression in mouse serum.It is speculated that acupuncture may improve the differentiation of Th1/Th2 cells and stimulate the inflammation of the knee joint by stimulating the expression of Treg cells.This may be part of the mechanism of acupuncture treatment of this disease.