| Objective:Using bovine type II collagen and Freund's complete adjuvant to establish collagen induced arthritis?CIA?rat model,to observe the effect of Huangqi Guizhi Wuwu Tang?HGW?on the rat model of arthritis index,joint pathology,toe swelling,blood rheology,apoptosis of synovial cells,cytokines and JAK3/STAT3 signaling pathways,to elucidate the molecular mechanism of its function.Investigate the effect of Huangqi Guizhi Wuwu Tang?HGW?on the mechanism of collagen induced arthritis in rats.Methods:Using the method of table of random number divided 72 SPF rats of SD were randomly into group?N?,model group?CIA?,Tofacitinib group?TFN?,HGW low,medium and high dose group?HGW-L,M,H?.HGW-L,HGW-M,HGW-H group of drug doses were 16,32,64 g·ml-1,equivalent to 3,6 ordinary clinical dosage,12 times,TFN dose was 15 mg·kg-1,distilled water group N,CIA were given 10 mg·kg-1.Using bovine type II collagen and Freund's complete adjuvant to establish collagen induced arthritis?CIA?rat model,measurement of rat lower limb volume by water volume method;The rat arthritis index was scored;watching the ankle pathological morphology in rats under the treatment of HGW after 4 weeks;TUNEL staining was used to detect the apoptosis of synovial cells;By using automatic blood rheometer of whole blood high shear and low shear of rats was detected in peripheral blood viscosity;The content of bovine type II collagen antibody,TNF-?,IL-1?,IL-6,CRP,RF and other cytokines were detected in the peripheral blood by ELISA;The content of peripheral blood IL-1a,IL-2,IL-10,IL-4,IFN-r and other cytokines were detected by flow cytometry;The protein expression level of VEGF,Bcl-2,BAX,SOCS1,SOCS3,STAT1,STAT3,JAK3 in rat synovial tissue were detected by immunohistochemistry;From the perspective of gene and protein to observe the expression levels of STAT3,JAK3 and its negative feedback regulatory factors SOCS1and SOCS3 to use Western blot technology.Results:Rats in group N had bright white hair color,good gloss,good eating,free movement of limbs,sensitive reaction,no swelling of paw,weight gain fast and stable;The hair of rats in group CIA were lackluster,their food intake was significantly reduced,their hind feet swelling ulceration with claudication;There is often a whining,like each other,arched back and curled up,food intake reduced,stool thin,unresponsive,weight gain is not obvious;The hair of color gloss of the rats in each medication group was improved;Their hind feet swelling andlimping were relieve;Activity increased,weight gain stable,the improvement of HGW-H group was better than that of other treatment groups.After modeling;The acute immune response of the rats with double full plantar and ankle joints;From the second week,compared with N group,CIA group,HGW-L,M,H group and TFN group rats toes swelling degree increased significantly?P<0.01?second and sixth weeks,compared with N group,CIA group,TFN group and HGW-L,M,joint inflammation of rats in the H group was significant difference?P<integral 0.01?.Compared with N group,the serum RF,CRP,CollagenII antibody levels in the CIA group were significantly different?P<0.01?,the difference was statistically significant.Under the light microscope,it was found that the synovial cells of N group had no regular proliferation,scattered blood vessels,no effusion in articular cavity,smooth joint surface.Synovial cell in CIA rats were hyperplasia,pannus formation,visible inflammatory cells,articular surface roughness,articular cartilage was severely damaged.TFN group and HGW-L,M,H after the drug group intervention,varying degrees of inhibition of the proliferation of synovial tissue,so that monocytes and lymphocytes are reduced in varying degrees.Compared with N group,the expression levels of STAT3 and SOCS1 in the CIA group and HGW-H group were significantly higher?P<0.05,P<0.01?;Compared with N group,the number of synovial cells in CIA group,HGW-L group,M group,H group,TFN group increased,apoptosis rate decreased significantly?P<0.05?;Compared with N group,The plasma viscosity and erythrocyte aggregation index?P<0.01?were significantly higher in the whole blood viscosity of rats in group CIA at 30/s,1/s and 200/s shear rate;Compared with group CIA,the content of HGW-H group and TFN group rats IL-4and IL-10 increased significantly?P<0.05,P<0.01?;compared with group N,the content of CIA in group IL-1?increased significantly?P<0.05?,IL-3,GM-CSF content decreased significantly?P<0.05?.Conclusion:Huangqi Guizhi Wuwu Tang has obvious therapeutic effect on Toe joint inflammation in CIA rat model,the mechanism may be that HGW plays a role of anti inflammation and immunity by regulating the content of inflammatory cytokines in peripheral blood of CIA rats.Huangqi Guizhi Wuwu Tang can promote the apoptosis of synovial cells,which may be one of the mechanisms in the treatment of rheumatoid arthritis.Huangqi Guizhi Wuwu Tang has anti-inflammatory and immunomodulatory effects through accommodating the protein in the pathway of JAK3/STAT3. |
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