| Objective:The research is based on the fact that "moxibustion has obvious anti-inflammatory effects of treating RA and cholinergic anti-inflammatory pathway can obviously inhibit systemic and local inflammation ". Guided by the theory of acupuncture and moxibustion, experimental RA rabbits were treated by moxibustion at "Shenshu"" Zusanli"points. Focusing on the key pathological character of synovitis, we selected cholinergic anti-inflammatory pathway as the breakthrough point of the research and "moxibustion stimulation-cholinergic anti-inflammatory pathway-anti-inflammatory effecf" as the main line of the research. By the use of nerve block, immunohistochemistry, RT-PCR and other methods and techniques, to explore cholinergic anti-inflammatory pathway’s mechanism of action in treating RA by moxibustion, the research is of important value for deepening the study of the principles of moxibustion treatment of RA and enriching scientific connotation of moxibustion.Methods:Japanese big-eared white rabbits were randomly divided into normal control group, RA model group, moxibustion treatment group, normal treatment of vagotomy group, moxibustion treatment of sham operation group. RA animal models were produced by Freunds1Complete Adjuvant (FCA) and left truncal vagotomy of the neck was as a means of intervention. RA rabbits were treated by moxibustion at "Shenshu"" Zusanli"points.1. RA rabbits’overall condition, body weight, knee joint perimeter, pathological morphology of synovial membrane tissue inflammation were observed.2. TNF-a, IL-1, IL-6, HMGB1in synovial fluid were detected by enzyme-linked immunosorbent assay (ELISA).3.Content of α7nAChR, ChAT in blood serum were detected by enzyme-linked immunosorbent assay (ELISA). Expression of a7nAChR, ChAT in synovial membrane tissue were detected by immunohistochemistry method and mRNA expression of a7nAChR, ChAT in synovial membrane tissue were detected by RT-PCR method.4. Expression of NF-KBp65, NF-κBp50, IκBα, p-IκBa, IKKβ in synovial membrane tissue were detected by immunohistochemistry method.Results:1. Moxibustion at "Shenshu" and "Zusanli" points of rabbits could obviously improve RA rabbits" overall condition, body weight, knee joint perimeter, pathological morphology of synovial membrane tissue inflammation, but its effect was decreased significantly after cutting off the vagus nerve.2. The expression results of cytokine in synovial fluid showed:Compared with the normal control group, RA model group’s expression of TNF a, IL-1, IL-6, HMGB1in synovial fluid were significantly increased (P<0.01); Compared with model group, moxibustion group and sham operation group were decreased significantly (P<0.01) and vagotomy group was not decreased significantly (P>0.05); Compared with moxibustion group, vagotomy group were significantly increased (P<0.01), but the sham operation group was not significantly increased (P>0.05).3. The expression results of a7nAChR and ChAT showed:Compared with the normal control group, RA model group’s expression of α7nAChR, ChAT in blood serum and proteinum expression of α7nAChR, ChAT in synovial membrane tissue mRNA expression of a7nAChR, ChAT in synovial membrane tissue were decreased significantly (P<0.01); Compared with model group, moxibustion group and sham operation group’s expression of a7nAChR, ChAT in blood serum and proteinum expression of a7nAChR, ChAT in synovial membrane tissue mRNA expression of a7nAChR, ChAT in synovial membrane tissue were significantly increased (P<0.01) and vagotomy group was not increased significantly (P>0.05); Compared with moxibustion group, vagotomy group were significantly decreased (P<0.01), but the sham operation group was not significantly decreased (P>0.05).4.①The expression results of NF-κBp65, NF-κBp50, p-IκBα, IKKα showed: Compared with the normal control group, RA model group’s expression of NF-κBp65, NF-κBp50, p-IκBα, IKKP in synovial membrane tissue were significantly increased (P<0.01); Compared with model group, moxibustion group and sham operation group’s expression of NF-κBp65, NF-KBp50, p-IκBα, IKKP in synovial membrane tissue were decreased significantly (P<0.01) and vagotomy group was not decreased significantly (P>0.05); Compared with moxibustion group, vagotomy group were significantly increased (P<0.01), but the sham operation group was not significantly increased (P>0.05).②The expression results of IκBα showed:Compared with the normal control group, RA model group’s expression of IκBαin synovial membrane tissue were significantly decreased (P<0.01); Compared with model group, moxibustion group and sham operation group’s expression of IκBα in synovial membrane tissue were increased significantly (P<0.01) and vagotomy group was not increased significantly (P>0.05); Compared with moxibustion group, vagotomy group were significantly decreased (P<0.01), but the sham operation group was not significantly decreased (P>0.05).Conclusions:1. Moxibustion at "Shenshu" and "Zusanli" points of rabbits can obviously improve RA rabbits’overall condition, knee joint perimeter,pathological morphology of synovial membrane tissue inflammation,which shows that moxibustion has a definite effect of anti-inflammatory, detumescence and inhibition of synovial inflammation. But its effect is decreased significantly after cutting off the vagus nerve. It prompts that anti-inflammatory effect of moxibustion for treatment of RA may be closely related to stimulating cholinergic anti-inflammatory pathway.2. Moxibustion at "Shenshu" and "Zusanli" points of rabbits can inhibit expression of TNF a, IL-1, IL-6, HMGB1in synovial fluid of RA rabbits, but its effect is decreased significantly after cutting off the vagus nerve. It prompts that stimulating cholinergic anti-inflammatory pathway may be one of the important mechanisms that moxibustion inhibit expression of proinflammatory cytokines in synovial fluid of experimental RA rabbits.3.Moxibustion at "Shenshu" and "Zusanli" points of rabbits can increase the expression of a7nAChR、ChAT in blood serum and synovial membrane tissue of RA rabbits, and activate the inherent cholinergic system in immune system, but its effect is decreased significantly after cutting off the vagus nerve. It prompts that moxibustion may promote the vagus nerve to release ACh and make ACh combine with a7nAChR of immune cells, thereby activating the inherent cholinergic system in immune cells and converting to diffuse, sustained immune regulation within the immune system.4. Moxibustion at "Shenshu" and "Zusanli" points of rabbits can inhibit the expression of NF-κBp65、NF-icBp50、p-IκBα、IKKP in synovial membrane tissue of RA rabbits,and increase the expression of IicBa, but its effect is decreased significantly after cutting off the vagus nerve. Therefore we speculate that by inhibiting the activation of IKK/IκB/NF-κB signal transduction pathway moxibustion may restrain high regulation of NF-κB in RA rabbits’synovium and inhibit the expression of Inflammatory cytokines in synovial cells, and then achieve its anti-inflammatory effect, in which cholinergic anti-inflammatory pathway plays an important role. |
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