Moxibustion Regulates The PD-1/PD-L1 Mechanism Of T Cell Function In Experimental RA Models

Objective:Under the guidance of acupuncture and moxibustion theory and focusing on the key pathological character of synovitis, we selected PD-1,PD-L1 as the breakthrough point of the research and "PD-1/PD-L1 inhibitory pathway-function of T lymphocyte-anti-inflammatory effect"as the main line of the study. By using immunohistochemistry, Elisa as well as other techniques to explore the effects of PD-1/PD-L1 inhibitory pathway in treating RA with moxa, the research aims at a further study of principles of moxibustion treatment.Methods:30 Japanese white rabbits were divided at random into the following three groups: normal control, RA model as well as moxibustion treatment group (n=10).RA model was established by injection of Complete Freunds' Adjuvant (CFA) into the articular cavities of bilateral hind limbs of rabbits. Moxa-cone moxibustion treatment was given to "Shenshu" and" Zusanli" points from the 8th day on after modeling, once daily for 3 weeks except Sundays. RA rabbits' overall condition, body weight, knee joint perimeter, pathological morphology of synovial tissue were observed; levels of IL-2, IFN-? and IL-17 in synovial fluid (SF) as well as p56lck and p59fyn in serum smaples were detected by ELISA; expressions of mPD-1 and mPD-L1 of splenic tissues were examined by immunohistochemistry, and levels of sPD-1 and sPD-L1 in sera and SF were detected by ELISA; NF-?Bp65 and IKK? in synovial tissue were examined by immunohistochemistry.Results:1. Moxibustion could obviously improve the animals' overall condition, gain their weight, reduce their knee joint perimeters, and enhance the effects of improving pathological morphology of inflammative synovium.2. The levels of IL-2, IFN-? and IL-17 in SF samples from the RA model group were elevated significantly as compared with those from controls (P<0.01); the levels of IL-2, IFN-? and IL-17 from the moxibustion group were reduced when compared with those from the RA model group (P<0.05).3. Compared with the normal control group, the levels of p56lck and p59fyn were elevated in sera of the RA model group (P<0.05); the levels of p56lck and p59fyn of moxibustion group were reduced when being compared with those of the RA model group (P<0.05).4. The expressions of mPD-1 and mPD-L1 in splenic tissues from the RA model group were elvevated in comparison with those from controls (.P<0.05);the expressions of mPD-1 from the moxibustion group were upregulated as compared with those from the RA model group(P>0.05), while the expressions of mPD-L1 were elvated(P<0.05).5. The levels of sPD-1 and sPD-L1 in sera and SF of the RA model were higher than those in the control group (P<0.05, P<0.01), and the levels of sPD-1 in the moxibustion group were lower than those in the RA model group(.P<0.05) while the levels of sPD-L1 were lower as compared with those in the RA model group(P>0.05).6. The expressions of NF-?Bp65 and IKK? in synovial tissue of RA models were significantly increased in comparison with those of controls (P<0.05); and the expressions of NF-KBp65 and IKK? in the moxibustion group were lower remarkably than in the RA model group (P<0.05).Conclusions:1. Moxibustion may have good therapeutic effects of anti-inflammatory, inhibition of synovial inflammation and cytokine secretion of T cells to experimental RA models.2. Moxibustion propably strengthen the signal transduction of PD-1/PD-L1 inhibitory pathway to regulate the overactivation of T cells.3. Moxibustion may regulate the overactivaton of NF-?B pathway in synovial cells which is associated clsosely with synovium inflammation by means of regulating T cell overactivation.