| Gout is an inflammatory disease caused by the deposition of uric acid crystals in the joints and surrounding connective tissue.The pathogenesis of gout is hyperuricemia(Hyperuricemia,HUA)caused by long term purine metabolic disorder and/or reduced excretion of uric acid(UA).The disease was characterized by recurrent hyperuricemia and acute characteristic arthritis at the beginning of the disease,and the deposition of sodium urate(monosodium utate calcium,MSU)was gradually developed.The disease can develop into chronic gouty arthritis,and may cause joint deformities,renal parenchymal lesions and urate formation.In China,the prevalence of acute gouty arthritis(AGA)has gradually increased and is becoming younger.With the development of immunology and molecular biology,it is found that the immune inflammatory injury induced by MSU is an important reason for the development of AGA,and the inhibition of joint immunological injury is a new target for the treatment of this disease.Our research team focused on the mechanism of HUA and gouty arthritis injury,and found that the NLRP3/NFkappa B signaling pathway mediated the inflammatory response in AGA.The AMPK/PPARgamma signaling pathway has the biological function of NLRP3/NF-kappa B.Enhancing the activity of AMPK/PPAR-gamma can effectively inhibit the immune inflammatory injury of joint tissues.Chinese herbal compound Tutengcao decoction is a prescription,which is created by my tutor,for the treatment of AGA.This paper will discuss my work in two parts:(1)Experimental Researches:In this study,with the starting of AMPK/PPAR-gamma and NLRP3/NF-kappa B signaling pathways,We observe the effects of Tutengcao on rat models of AGA and explore the the molecular mechanism of suppressing the immune inflammatory damage of AGA model rats.(2)Clinical researches:Analysising the effectiveness and safety of Tutengcao by observing the effeccts of Tutengcao on clinic symptom,syndrome and laboratpry tests of AGA patiets.1 Experimental ResearchObjective:Exploring the Tutengcao’s mechanism of inhibiting the immune inflammatory injuries in AGA by observing the degree of joint swelling,inflammation index,dysfunction score,serum biochemical index,CRP and AMPK/PPAR-gamma,NLRP3/NF-kappa B signaling pathway genes and protein expression in serum and synovium tissue of rat models with AGA.Methods:Healthy male SD rats were gavaged with adenine and fed with yeast for 21 days and the sUA were detected.Models were regarded as successful hyperuricemia models if the sUA>110mmol/L.The rats were injected with 0.2 ml and 0.1ml of MSU solution in the knee and ankle of the right hind limb respectively.Then the joints were swollen,which was considered as successful AGA model.Thirty male rats were randomly divided into model group,colchicine group(0.3mg·kg-1·d-1),large and medium-dose group of Tutengcao granuless(1、0.5、0.25g·kg-1·d-1),6 rats in each group,and 6 rats in normal group.The ankle joint thickness was measured and the sacroiliac joint inflammation index and dysfunction were evaluated daily.One week after drug intervention,collect the rats’ 24 urin and detect 24-hour urine protein.After that,the rats were sacrificed and serum,synovial tissues were taken.Detect the serum levels of UA,Hcy,CRP,BUN,Cr.The mRNA levels of AMPKα2,PGC-1α,PPAR-γ,NLRP3 and NF-κB P65 in synovial tissues were detected by RT-PCR.Western blot was used to detect the expression of AMPKα2,pAMPKα2,PGC-1α,PPAR-γ,NLRP3 and NF-κB P65 in synovial tissues.Results:①Joint symptoms:Right ankle joint width:Compared with the model group,the thickness of the sacroiliac joint of positive drug group and the large,medium and low doses of Tutengcao(TTC)granules decreased in the 2nd d and 3thd(P<0.05,P<0.01).Inflammatory index of joint:The inflammatory index of the sacroiliac joints of the rats in the positive drug group,the large,medium and low doses of TTC granules decreased in the 1st and 2nd d(P<0.01 or P<0.05).The score of joint dysfunction:The scores of rats’ ankle joint dysfunction in the large,medium,and low doses of TTC granules decreased(P<0.01 or P<0.05).E② Biochemical results:Compared with the normal group,the sUA of the model group increased before and after treatment(P<0.01).After 1 week treatment,compared with the normal group,CR,BUN and uPRO/24h increased in the model group(P<0.01).Compared with the model group,the CR and uPRO/24h of the positive drug group and all groups of Chinese medicine decreased(P<0.01 or P<0.05),and the positive drug group,the BUN of the high dose group of TTC granules decreased(P<0.05).Compared with the normal group,TG and Hey increased in the model group(P<0.05).Compared with the model group,TG in the positive drug group decreased(P<0.05),and the Hey in the low dose group decreased(P<0.05).③CRP:After 1 week treatment,compared with the normal group,the serum CRP of the model group showed an upward trend.④ RT-PCR results:Compared with the normal group,AMPK alpha 2,PGC-1 alpha and PPAR-gamma mRNA in synovial membrane of model group were down regulated and NLRP3、P65 mRNA were up regulated(P<0.01,P<0.05).Compared with the model group,the AMPK alpha 2,PGC-1 alpha and PPAR-gamma mRNA of synovial membrane in the positive drug group were up regulated(P<0.05),the PGC-1 alpha mRNA in the high-dose TTC granules group was upregulated(P<0.01).Compared with the model group,the NLRP3 mRNA was down regulated in the positive drug group and the medium,low-dose TTC granules groups(P<0.05),while the P65 mRNA in the positive drug group,the high,low-dose TTC granules groups were down regulated(P<0.05).⑤Western blot results:Compared with the normal group,the expression of AMPK alpha 2,pAMPK alpha 2,PGC-1 alpha and PPAR-gamma in the synovial membrane of the model group was down regulated,and the expression of NLRP3 and P65 protein was up regulated(P<0.01)(P<0.01).Compared with the model group,the expression of AMPK alpha 2 and pAMPK alpha 2 protein in each treatment group was up regulated,and the expression of P65 was down(P<0.01,P<0.05).The expression of PGC-1 alpha protein in the positive drug group and the medium dose TTC granules group was up regulated(P<0.05),and the expression of PPAR-gamma protein in the medium dose TTC granules group was up regulated(P<0.05).Compared with the model group,the expression of NLRP3 protein in the positive group,the medium,low dose TTC granules groups were down regulated(P<0.05).Conclusion:TTC granules can effectively improve joint swelling,inflammation index and dysfunction score of AGA model rats,as well as reducing CRP,CR、BUN、uPRO/24h.The molecular mechanism of protecting rat synovial tissue may be related to the activation of AMP K/PPAR-gamma,thereby inhibiting the activity of NLRP3/NF-B and reducing the inflammatory reaction.2.Clinical ResearchObjectiveTo observe the curative effect and safety of Tutengcao Decoction(TTCD)in treating patients with acute gouty arthritis(AGA).Methods:We enrolled 121 acute gouty arthritis patients with hyperthermia,phlegm,and internal resistance from October 2016 to January 2018 in Dongfang Hospital,and patients were randomly assigned in control group and treatment group.The control group received the loxoprofen sodium tablets,while the treatment group was treated with TTCD.The treatment time was 1 week.The safety was evaluated during the treatment time,and the curative effect was also evaluate after 1week by sUA、ESR、CRP、CRE、BUN、Pro and clinical symptoms,the length of time required for the remission of joints.Results:We enrolled 121 cases in the early stage,and the19 of them were shedding.In the end,we actually enrolled 102 cases,of which 50 cases in the control group,52 cases in the treatment group.The graner,age,history of alcohol and smoking and disease condition of the two groups were comparable.After one week,all the tagets were examined.After 1 week treatment,compared with before treatment,CRP,ESR,sUA and Hcy decreased after treatment(P<0.01).Compared with the control group,the level of sUA in the treatment group decreased(P<0.05).Compared with the control group,the pain and swelling relief time in the treatment group showed a downward trend(P>0.05),and the time of joint dysfunction relief was shortened(P<0.05).Compared with those before treatment,the two groups of joint swelling and pain scores,VAS scores,TCM symptoms and signs scores were all decreased(P<0.01).Compared with the control group,those scores in the treatment group decreased(P<0.05).In the improvement of acute renal injury,the improvement rate of Pro in the treatment group was significantly higher than that in the control group(P<0.05).Conclusion:Treatment of acute gouty arthritis with Tutengcao Decoction played a better role in improving AGA clinical symptoms,acute kidney injuries,and reducing sUA than loxoprofen sodium. |
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