Multicenter Clinical Observation And Experimental Study On The Treatment Of RA With Yunpi Jiedu Tongluo Qushi Granules

Objective:A prospective cohort study was used to include 312 patients with rheumatoid arthritis with spleen deficiency and dampness,toxicity and stasis paralysis evidence in 16 centers,with the Yunpi Jiedu Tongluo Qushi Granules with identification and addition of flavor as an exposure factor.To observe the clinical efficacy and safety of spleen deficiency and dampness,toxicity and stagnation paralysis in rheumatoid arthritis by combining methotrexate with Yunpi Jiedu Tongluo Qushi Granules in a clinical trial;to investigate the mechanism of Yunpi Jiedu Tongluo Qushi Granules in CIA model rats through animal experiments.Methods1.Clinical observation:a prospective cohort study was used to include 312 patients with rheumatoid arthritis with spleen deficiency and dampness,toxicity and stasis paralysis evidence in 16 centers,with the application of Yunpi Jiedu Tongluo Qushi Granules as an exposure factor.Among them,166 cases in the exposed group were treated with the Yunpi Jiedu Tongluo Qushi Granules and Western medicine;146cases in the non-exposed group were treated with Western medicine alone(with methotrexate as the basic drug or in combination with/without other traditional anti-rheumatic drugs to improve the disease)for a total of 24 weeks of intervention.Visits were conducted at three time points:week 0,12 and 24 weeks.Observed indicators:primary outcome indicators were ACR response criteria and secondary outcome indicators were 28 joint mobility(DAS28)score,DAS28 remission rate,clinical disease activity index,visual analogue score of pain,TCM symptom score,health assessment questionnaire,FACIT-Fatigue scale,dynamic erythrocyte sedimentation rate,C-reactive protein,rheumatoid factor,antiexchanging citrullinated peptide antibody and adverse reactions.2.Animal experiments:25 male SD rats were selected and divided into 5 groups by random number table method,5 rats in each group,divided into blank group,model group,MTX group,spleen detoxification,Tongluo dispelling dampness pellet group,and spleen detoxification,Tongluo dispelling dampness pellet+MTX group.Type Ⅱcollagen emulsifier was used for modeling,and 0.2ml of 1 mg/ml immunogen was subcutaneously injected into one side of the tail root of selected SD rats,and 0.15ml of 1mg/ml immunogen was injected into the other side of the tail root of the rats again one week later to strengthen immunization and establish the CIA rat model.The rats were administered by gavage on the day of the first immunization injection until the material was taken,while the blank group and the CIA model group were given only equal amounts of saline.The rats were continuously monitored for weight change and joint swelling during feeding.The rats were anesthetized with 10%chloral hydrate intraperitoneally and then blood was removed from the abdominal aorta.The case pictures of the knee joints of the hind limbs of rats were examined after sampling to observe the local inflammation,inflammatory cell infiltration and bone erosion;Elisa was used to detect the CRP and anti-CCP antibodies in the serum of rats;the expression of m6A reading protein YTHDF2 in the replacement joints of rats in each group was detected by Western Blot method,and the preliminary exploration of the effects of spleen transporting and detoxifying Tongluo dispelling particles on the inflammatory indexes of CIA rats and YTHDF2 protein in CIA rats.Results1.Clinical study Clinical efficacy evaluation:the results of the pilot study showed that the exposed group had a higher percentage of meeting the improvement criteria of ACR20,ACR50 and ACR70 at 12 weeks,a higher remission rate of DAS28 than the non-exposed group,a lower CDAI,number of joint swelling,number of joint pressure pain,and Chinese medicine evidence score than the non-exposed group,and a higher FACIT-F score than the non-exposed group(P<0.05).In the exposed group,the CDAI score was lower than that of the non-exposed group and the FACIT-F score was higher than that of the exposed group at 24 weeks(P<0.05).In the subgroup analysis using MTX only,at week 12,the exposed group had lower DAS28,joint pressure pain,and Chinese medical evidence scores and higher FACIT-F scores than the nonexposed group(P<0.05);at week 24,the exposed group had lower DAS28,CDIA,joint pressure pain,and Chinese medical evidence scores and lower DAS28 remission rate,DAS28 hypermobility rate,and FACIT-F scores were higher than those of the non-exposed group,with statistically significant differences(P<0.05).Combined medication analysis:the exposed group had less combined medication than the non-exposed group at 24 weeks(P<0.05),and the combined medication of DMARDs and NSAIDs was improved in the non-exposed group(P<0.05)Analysis of adverse reactions and safety indicators:the incidence of gastrointestinal adverse reactions,bone marrow suppression,abnormal liver and kidney function and other adverse reactions were lower in the exposed group than in the non-exposed group during treatment(P<0.05),and the HGB was elevated at 12 and 24 weeks compared with the nonexposed group(P<0.05).2.Animal experiments1 The spleen detoxification and dampness dispelling granules can reduce joint symptoms and promote weight gain in CIA model rats(P<0.05),protect synovial cell morphology and reduce inflammatory cell infiltration.The spleen detoxification,Tongluo dispel dampness granules can reduce the serum content of CRP and anti-CCP antibodies in CIA model rats(P<0.05).2 Yun spleen detoxification,Tongluo dispel dampness granules could increase the expression of m6A methylation recognition protein YTHDF2 protein in the ankle joint of CIA rats(P<0.05).Conclusions1 Conclusions of clinical study The combination of spleen detoxification and Tongluo dispelling dampness granules with Western medicine treatment can improve the remission rate of ACR20,50,70 and DAS28 at 12 weeks,reduce the CDAI score,the number of joint swelling,the number of joint pressure pain in subjects,ESR and improve the overall disease status;reduce the Chinese medicine evidence score in subjects,improve the FACIT-F score in subjects,and improve the patients’ living status.The study was carried out on the basis of the same ACR improvement rate and DAS28 score,to reduce the incidence of combined medication and adverse reactions.2 Conclusions of animal experimentThe spleen-relief and detoxification Tongluo dispel dampness particles can reduce joint symptoms,promote weight gain,protect synovial cell morphology,reduce inflammatory cell infiltration,reduce CRP and anti-CCP antibodies in the serum of CIA model rats,and increase the expression of m6A methylation recognition protein YTHDF2 protein in the ankle joint of CIA rats.