| Objective: To observe the effect of Zhuang medicine Longzuantongbi granules on cytokines MEK1/2 and ERK1/2 in serum,synovium and cartilage tissue of rats with rheumatoid arthritis,as well as the effect on pathological changes of synovium tissue of joints,discusses Zhuang medicine Longzuantongbi granules through the side of the mechanism of action of rheumatoid arthritis.For the clinical application of Zhuang medicine Longzuantongbi granules and effective prevention and treatment of rheumatoid arthritis provide reliable evidence.Methods: A total of 96 SD rats were selected and 24 were randomly selected as the normal control group(the normal group).The remaining 72 were copied by the combined modeling method of wind,cold and wet environmental factors combined with biological factors.After successful replication of the RA model,they were randomly divided into three groups:model group,western medicine group and zhuang medicine group,with 24 in each group.The total daily gavage volume of each rat was set at 1ml/100 g.Normal group and model group were given normal saline gavage once a day.The western medicine group was given methotrexate solution 0.9mg/kg once a week,and normal saline was given for the remaining six days.Zhuang medicine group was given 0.27g/kg of Zhuang medicine Longzuantongbi granules solution every day,and was administered in two times.Specimenswere collected at the 7th,14 th,21st day after administration and at the 1st week after withdrawal(the 28 th day).According to the sampling time,each group was divided into four subgroups: 7d,14 d,21d and 28 d.The whole experimental rats were divided into 16 groups.Bone tissues of the ankle joint and below were stained with HE,and then pathological sections were made.After the whole blood serum was collected from the rats,the protein contents of MEK1/2 and ERK1/2 in the serum of each group were detected by enzyme-linked immunoassay(ELISA),and the protein and gene expression levels of MEK1/2 and ERK1/2 in the synovial tissues were detected by Western Blot and RT-PCR.SPSS 25.0 statistical software was used for data analysis.Result: 1.Results of HE: according to the observation of the pathological sections of the ankle joint in each group on the 21 st day after the administration of the drug,the synovium of the ankle joint in the normal group was smooth without hyperplasia,the distribution of chondrocytes was regular and clear,and no inflammatory infiltration was observed in the connective tissue around the joint.Compared with the normal group,the synovium surface of the model group was less smooth,with obvious hyperplasia,narrowing of the articular cavity,and the distribution of a large number of lymphocytes.Compared with the model group,both the western medicine group and the zhuang medicine group showed less inflammatory infiltration after administration,with more uniform distribution and orderly arrangement of chondrocytes,less lymphocytes,less inflammatory exudation,less synovial membrane,and smooth surface.2.ELISA method results: the protein contents of MEK1/2,ERK1/2 and p-ERK in the serum of normal group,model group,western medicine group and zhuang medicine groupwere not statistically significant(P > 0.05).3.Western Blot test results:compared with the normal group,the expression levels of MEK1/2,ERK1/2and p-ERK in the synovial tissues of the model group were up-regulated,and the differences were statistically significant(P < 0.05).Compared with the model group,the expression of synovium protein was down-regulated in the zhuang medicine group and the western medicine group,and the difference was statistically significant(P < 0.05).There was no significant difference between the western medicine group and the zhuang medicine group(P >0.05).4.Real-time fluorescence PCR results: The m RNA expression of MEK1/2 and ERK1/2 was up-regulated in the model group compared with the normal group(P < 0.05).Compared with the model group,the m RNA expressions of MEK1/2 and ERK1/2 in the zhuang medicine group and the western medicine group were close to those in the normal group,and the difference was statistically significant(P < 0.05).There was no significant difference between the zhuang medicine group and the western medicine group(P > 0.05).Conclusions: By down-regulating the gene and protein expression of MEK1/2 and ERK1/2 in the joint tissues of rheumatoid arthritis model rats,Zhuang medicine Longzuantongbi granules may reduce synovial lesions,delay the process of cartilage and bone destruction,and achieve the therapeutic effect on rheumatoid arthritis. |
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