Mechanisms Of Zhengqing Fengtongning And Methotrexate Therapy On Bone Destruction Of Rheumatoid Arthritis

Objective:1 To investigate the effect of Zhengqingfengtongning(ZQ) and methotrexate (MTX), used alone or in combination, on bone erosions of RA;2 To approach the possible mechanisms of ZQ and MTX on bone erosions from the aspect of inflammation and RANKL system.Methods:1 Synovial cell was incubated in vitro. These cells were divided into 3 experimental groups (MTX group (0.1mg/ml), ZQ group (0.1mg/ml), ZQ+MTX group (0.1mg/ml ZQ+ 0.1mg/ml MTX) and 1 control group. The cultured cells were observed by light microscope and fluorescence microscope, the flow cytometer were used to detect apoptosis of Synovial cell, and the expression of RANKL and OPG mRNA were detected by RT-PCR.2 The rats in which CIA were divided into 4 groups:the control group, the model group, the MTX group (0.1mg/ml), ZQ+MTX group (0.1mg/ml ZQ+0.1mg/ml MTX). Arthritis were evaluated by arthritis index and the expressions of OPG,RANKL,IL-1α,IL-6,MMP-3 and MMP-13 in serum were detected by ELISA.Results:1 HE staining, light microscopy:apoptotic cells were round, deeply stained nuclei, cytoplasm condensed, and chromatin into a mass; fluorescence microscopy:nuclear morphological changes, with formation of apoptosis bodies.2 By means of the flow cytometry to test the apoptosis rate:Control group25.3%, MTX group28.4%, ZQ group32.3%, Joint group34.7%. It showed that the apoptosis rate were increased in experimental groups, compare with control group (P<0.05).3 It was found that the expression of RANKL mRNA in experimental groups can be decreased, while OPG mRNA be increased, compare with control group (P<0.05).4 It was found that ZQ and MTX can suppress significantly the secondary paw swelling and pain response, as well as down-regulated the index of poly arthritis in CIA rats(P≤0.05).5 It was found that the expression levels of IL-lα, IL-6, RANKL, MMP-3 and MMP-13 in Joint group and MTX group were significantly lower than model group (P≤0.05).6 It was found that Joint group can reduce the expression levels of RANKL in serum, enhance the expression levels of OPG, increase OPG-RANKL ratio (P≤0.05).Conclusion: 1 ZQ and MTX can inhibit the proliferation of synovial cells, and affect the balance of OPG-RANKL system regulation.2 ZQ and MTX can control the joint swelling, and increase OPG-RANKL ratio.3 ZQ and MTX can protect the decreased bone density. This protective effect would be related to the inhibiting synovial cells growth and modifying the balance of OPG-RANKL.