The Research Of Zhuang Medicine Longzuantongbi Decoction On Regulation Effect Of Rheumatoid Arthritis Rat Models RANK/RANKL/NF-?B Signal Pathway

Objective: To observe the impact of Zhuang medicine Longzuantongbi Decoction on rheumatoid arthritis rat models serum Cytokine interleukin 1 ?(IL-1?)?tumor necrosis factor(TNF-?)?RANKL?RANK and NF-?B in synovium,and cells imbalance of Th17/Treg,try to discuss the mechanisms of Zhuang medicine Longzuantongbi Decoction regulating Rheumatoid Arthritis correlation factor in RANK/RANKL/NF-?B signal pathway and cellular immunity.Methods: Randomly selected 12 from 72 Male SD rats weight(120±30)g to normal control group(blank group),the rest of 60 were15 °slantways pricked into the toe from right hind limbs using NO.6needle,0.1ml complete Freund's adjuvant was intradermal injected to cause inflammation,at the seventh day did one more time in the way above,to establish models of RA.After that we randomly divided the rats into five groups:12 in model group,12 in methotrexate group,12 in each high,intermediate and low Zhuang medicine Longzuantongbi Decoction groups.The blank group and model group were given normal saline intragastric administration according to the dose of 2m L/ 100 g,once a day;methotrexate group were given methotrexate intragastric administration according to the dose of2.7mg/kg,once a week;high,intermediate and low Zhuang medicine Longzuantongbi Decoction groups had accepted intragastric administration of high,intermediate and low Zhuang medicine Longzuantongbi Decoction according to the dose of 6g/100g/d ?3g/100g/d ? 1.5g/100g/d respectively,twice a day,treated for 21 consecutive days in all.On the 21 th day,we collected blood from aortaventralis of the rats,collected the synovium from their keen joints.We use Enzyme linked immunosorbent assay to detect the level of serum IL-1? ? TNF-? in each group of rat models;use RT-PCR ?immunohistochemical technique to detect the expression of RANK?RANKL and NF-?B in synovium of RA rats,use fluid cytology test to detect the level of serum Th17 and Treg;use SPSS 19.0 statistical software for data processing.Results: 1.the expression level of serum IL-1??TNF-?:the level of serum IL-1??TNF-? in each treatment group were all lower than model group,the difference had statistical significance(P<0.05);compare with methotrexate group,the level of serum IL-1? ? TNF-? in high Zhuang medicine Longzuantongbi Decoction groups obviously decreased,the difference had statistical significance(P<0.05);compare with low Zhuang medicine Longzuantongbi Decoction group,the level of serum IL-1? ? TNF-? in high and intermediate Zhuang medicine Longzuantongbi Decoction groups obviously decreased,the difference had statistical significance(P<0.05).2.the expression level of RANK?RANKL?NF-?B gene in synovium:the expression level of RANK?RANKL?NF-?B gene in synovium in each treatment group were all lower than model group,the difference had statistical significance(P<0.05);compare with low Zhuang medicine Longzuantongbi Decoction group,the level of RANK ? RANKL ? NF-?B gene in high and intermediate Zhuang medicine Longzuantongbi Decoction groups obviously decreased,the difference had statistical significance(P<0.05).3.the expression level of RANK ? RANKL ? NF-?B protein in synovium:the expression level of RANK?RANKL?NF-?B protein in synovium in each treatment group were all lower than model group,the difference had statistical significance(P<0.05);compare with low Zhuang medicine Longzuantongbi Decoction group,the level of RANK?RANKL?NF-?B protein in high and intermediate Zhuang medicine Longzuantongbi Decoction groups obviously decreased,the difference had statistical significance(P<0.05).4.the expression level of serum Th17?Treg:compare with blank group,the level of serum Th17 in model group obviously increased,the level of serum Treg obviously decreased,the difference had statistical significance(P<0.01);compare with model group,the level of serum Th17 in each treatment group obviously decreased,the level of serum Treg obviously increased,the difference had statistical significance(P<0.05);compare with low Zhuang medicine Longzuantongbi Decoction group,the level of serum Th17 in high and intermediate Zhuang medicine Longzuantongbi Decoction groups obviously decreased,the level of serum Treg obviously increased,the difference has tatistical significanceConclusion(s):1.Zhuang medicine Longzuantongbi Decoction can decrease rheumatoid arthritis rats serum IL-1??TNF-? levels;2.Zhuang medicine Longzuantongbi Decoction can decrease rheumatoid arthritis rats level of RANK ? RANKL and NF-?B in synovium,improve the inflammation symptoms of join;3 ? Zhuang medicine Longzuantongbi Decoction can decrease Th17 and increase Treg,to promote cell immune regulation.