| Objective Gouty Arthritis（GA）model rats were treated with the traditional Chinese medicine Huashi Tongluo Jiangzhuo Recipe,and then the general condition,joint swelling index,pathological sections of the rats were observed,combined with serum and joint synovial tissue protein and gene detection,to explore the anti-inflammatory effect of traditional Chinese medicine Huashi Tongluo Jiangzhuo Recipe on GA rats and its effect on TLRs/NF-kB pathway.Methods: In this experiment,60 male SD rats with SPF grade and body weight of200g（±20g）were selected and randomly divided into blank group,model group,colchicine group and Huashi Tongluo Jiangzhuo prescription high,medium and low A total of 6 groups,10 rats in each group,were administered adaptively for a total of7 days.After intragastric administration on the 5th day,the diameters of the ankle joints of all rats were measured and recorded with a digital caliper.The GA model was established by injecting MSU suspension into the ankle of the rats in the high,medium and low groups of Tongluo Jiangzhuo Formula.After the model was successfully replicated,the rats were given intragastric administration for two days.Hour ankle diameter.On the 7th day after gavage,all rats were fasted for 24 hours,and abdominal aortic blood was collected to detect the protein levels of IL-1β,IL-6,and TNF-α in serum by ELISA.TLR2,TLR4,my D88,NF-κB m RNA expression in synovium was detected by Western-blot method,TLR2,TLR4,my D88,NF-κB protein expression in synovial tissue was detected by Western-blot method;ankle joint tissue was taken and HE staining was used to observe the disease.Physiological change.Methods In this experiment,60 male SD rats with SPF grade and body weight of 200g（±20g）were selected and randomly divided into blank group,model group,colchicine group,and Huashi Tongluo Jiangzhuo Fang（high,medium and low）.group,10 rats in each group,adaptive administration for a total of7 days,after gavage on the 5th day,the diameter of the ankle joints of all rats was measured and recorded with a digital tape caliper,model group,colchicine group,Huashitongluo Jiangzhuofang high,medium and low groups were injected with MSU suspension into the ankle joint to replicate the GA model.After the model was successfully replicated,the rats were given intragastric administration for two days.During this period,the ankles were recorded at 6,12,24,48,and 72 hours after modeling.Joint diameter.On the 7th day after gavage,all rats were fasted for 24 hours,and abdominal aortic blood was collected to detect the protein levels of IL-1β,IL-6,and TNF-α in serum by ELISA.TLR2,TLR4,my D88,NF-κB m RNA expression in synovium was detected by Western-blot method,TLR2,TLR4,my D88,NF-κB protein expression in synovial tissue was detected by Western-blot method;ankle joint tissue was taken and HE staining was used to observe the disease.Physiological change.Results（1）General conditions of rats: Compared with the normal group,the model group,colchicine group,and high-medium-low Chinese medicine group had lower scores of hair,nails,food intake,and spirit,among which the model group and the high-medium group were the most obvious.（2）HE staining of rat ankle joint: the results are shown.In the normal group,the ankle joint surface was continuous and smooth,the structure was complete,the synovial cells were arranged in a columnar and orderly manner,and there was no synovial hyperplasia and inflammatory cell infiltration;in the model group,the synovial surface was significantly proliferated,the synovial cells were arranged disorderly,and the articular surface was no longer smooth.,the structure of the joint cavity was destroyed,accompanied by inflammatory exudates;no obvious lesions were found in the colchicine group and the middle and low dose groups of Huashi Tongluo Jiangzhuo Recipe,indicating that the inflammation was controlled;the synovial membrane of the high dose Huashi Tongluo group was Mild hyperplasia and a little exudation of inflammatory substances appeared,and the articular surface was slightly rough,which may be related to the excessive concentration of the drug,which affected the absorption.（3）ELISA detection of rat serum: The results showed that IL-1β,IL-6 and TNF-α proteins in each group were higher than those in the blank group,while the increase in the colchicine group and the middle and low dose groups of traditional Chinese medicine was relatively small.significant difference.（4）TLR2,TLR4,my D88,NF-κB gene detection in rats,the results showed that compared with the normal group,the model group had the strongest expression,which was significant.The colchicine group and the low-dose Chinese medicine group were compared with the model group The difference was the largest,and there was no significant difference between groups.（5）WB detection of rat knee joint synovium: The results showed that the protein expression levels of TLR2,TLR4,My D88 and NF-κB in the model group,colchicine group and Chinese medicine high,medium and low dose groups were significantly higher than those in the blank group,indicating that TLR2,TLR4 And My D88,NF-κB showed high expression in the pathogenesis of GA.Conclusion 1.Huashi Tongluo Jiangzhuo Recipe can significantly reduce the inflammatory response in GA rats;2.The mechanism of Huashi Tongluo Jiangzhuo Recipe reducing inflammatory response may be related to the regulation of TLRs/NF-kB signaling pathway. |
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