The Cellular And Molecular Immunoregulatory Mechanisms Of Acupuncture On Collagen-induced Arthritis In Cx43 Knockout Mice

PartⅠEstablishment of Collagen-induced Arthritis model in Cx43 knockout MiceObjectives To establish the animal model with collagen-induced arthritis(CIA) using Cx43 knockout mice.Methods The mice were immunized by intradermal injection of bovine collagen typeⅡ(BCⅡ) emulsified with complete Freund's adjuvant(CFA). The swelling of feet were observed. The pathological characteristics of the animal model were assayed by photos of X-ray, and histopathological techniques. The level of anti-CⅡantibody in serum was analyzed by enzyme linked immunosorbent assay(ELISA).Results Obvious swelling of the feet was found in the model group compared with control group(P<0.05). Anti-CⅡantibody level was higher in model group than that in control. Besides, typical arthritis pathology was showed by roentgenography and histopathology.Conclusion The model of CIA in Cx43+/+ and Cx43+/- mice is established successfully, which is benifical to investigate pathogenesis and the therapy of human RA and Cx43-related research. The mice aged 9 weeks in Cx43+/+ and Cx43+/- are much easier to development CIA, and the former is much easier to development CIA.PartⅡEffect of Acupuncture on Th1/Th2 balance in Cx43 knockout mice with CIAObjectives To explore the correlation of Cx43 and efficacy of acupuncture, by observing the efficacy of acupuncture on CIA mice genotyped Cx43 +/+ and Cx43+/- , and observing the splenic Th1/Th2 cell subsets.Methods Acupuncture treatment,begun on four weeks after the initial injection of collagen, was given once a day for 3 weeks continuously. The arthritic index was observed. The intracellular cytokine detection was performed with three-color flow cytometry to quantitate the splenic Th1/Th2 cell subsets in mice with CIA induced by BCⅡand the sham-immunized controls.Results The arthritic index decreased significantly in Cx43+/+ mice with CIA compared with the Cx43+/- ones after acupuncture treatment. Acupuncture suppresses the Th1 subset and Th1/Th2 in Cx43+/+ mice. Whereas it has no obvious effect on Th2. In contrast, It has no effect on Th1 and Th2 in Cx43+/- mice with CIA.Conclusion Acupuncture has better efficacy on CIA mice genotyped Cx43+/+ than ones genotyped Cx43+/-. It showed the efficacy of acupuncture is associated with Cx43. Its active mechanism might decrease Th1 and regulate the proportion of Th1/Th2. There is a positive correlation among the effect of acupuncture and the regulation. PartⅢEffect of Acupuncture on IL-1βand TNF-αin Cx43 knockout mice with Collagen-induced ArthritisObjectives To explore the correlation of Cx43 and efficacy of acupuncture by observing the efficacy of acupuncture on CIA mice genotyped Cx43 +/+ and Cx43+/-, and observing the level of IL-1βand TNF-αin serum.Methods Acupuncture treatment,begun on four weeks after the initial injection of collagen, was given once a day for 3 weeks continuously. The arthritic index was observed. ELISA method was used to examine the levels of serum IL-1βand TNF-α.Results The arthritic index decreased significantly in Cx43+/+ mice with CIA compared with the Cx43+/- ones after acupuncture treatment. The level of IL-1βand TNF-αin serum decreased significantly in Cx43+/+ mice with CIA. Whereas there is no obvious change in Cx43+/- mice with CIA.Conclusion Acupuncture has better efficacy on CIA mice genotyped Cx43+/+ than ones genotyped Cx43+/-. It showed the efficacy of acupuncture is associated with Cx43. To decrease level of IL-1βand TNF-αis the possible mechanism of acupuncture to treat RA. There is the positive correlation among the effect of acupuncture and the regulation.