Therapeutic Effect of Total Triterpenoids of Chaenomeles speciosa Combined with Indomethacin on Rheumatoid Arthritis Induced by Freund's Complete Adjuvant in Rats
Objective:To observe the therapeutic effect of total triterpenoids of Chaenomeles speciosa(C.speciose)combined with indomethacin on adjuvant arthritis(AA)rats,and explore its possible mechanism.Methods;Rats were randomly divided into normal control group,model control group,indomethacin(5 mg/kg)group,total triterpenoids of C.speciose(100 mg/kg)group and combination group(total triterpe-noids of C.speciosa 100 mg/kg and indomethacin 5 mg/kg).Except for the normal control group,rats in each group were subcutaneously in-jected with Freund's complete adjuvant(CFA)on the right hind foot.After 14 days,corresponding medicine was administered by gavage once a day for 28 consecutive days,and foot swelling and arthritis index scores were calculated.The next day after the last administration,blood and synovial tissue of ankle joint were collected,and reactive oxygen species(ROS),lactate dehydrogenase(LDH)and endogenous antioxi-dant enzymes in serum,inflammatory factors in synovial tissues,the activities of cathepsins B and D,β-glucuronidase,β-galactosidase in syno-vial tissues and lysosome,and the histological changes of synovial tissues were determined and analyzed.Real-time PCR and Western blot were employed to detect the mRNA and protein expressions of related genes to TLR4/NF-κB/NLRP3 signaling pathway.Results:Compared with the normal control group,the model control group presented increased foot swelling and arthritis index scores(P＜0.01),cartilage de-struction and inflammatory infiltration,and up-regulated ROS,lipid peroxidation(MDA),myeloperoxidase(MPO),LDH in serum,elevated cyclooxygenases-2(COX-1),COX-2,prostaglandin E2(PGE2),IL-1β,TNF-α,IL-6,IL-18,cathepsins B and D,β-galactosidase,β-glucuroni-dase in synovial tissues,and enhanced activities of β-galactosidase and cathepsin D in the cytoplasm of lysosomal organelles(P＜0.01).Ad-ditionally,the model control group had a decrease in superoxide dismutase(SOD),catalase(CAT),and glutathione peroxidase(GSH-Px)in serum,IL-4 and IL-10 in ankle joint tissue and the activities of β-glucuronidase and cathepsin D in lysosomal organelles(P＜0.01).There was an up-regulation of mRNA in Tlr4,Myd88,Nlrp3,Asc,Caspase 1,Cox 1,Cox2,Pge2 and protain in p-IκBα,p-IKKβ,TXNIP,NEK7,NL-RP3,ASC,Pro-caspase-1,Caspase-1,GSDMD,GSDMD-N,Pro-IL-1β,and ProIL-18,nuclear NF-KBp65,p-IKBα/IKBα and p-IKKβ/IKKβ in synovial tissues(P＜0.01),while a down-regulation in cytosol NF-KBp65 in synovial tissues(P＜0.01).Compared with the model control group,the combination group had decreased foot swelling and arthritis index score(P＜0.01),reduced cartilage destruction and inflammatory infiltration,and increased COX-1 activity and mRNA expression(P＜0.01).The ROS,LDH,oxidative stress indicators in serum,cytokines in synovial tissues,the activity of lysosomal enzymes in synovial tissues,organelles and cytoplasm of lysosomal,the mRNA and protein expres-sions of related genes to TLR4/NF-κB and NLRP3 signaling pathways in synovial tissues were reversed(P＜0.01).Conclusion:The combi-nation of total triterpenoids of C.speciosa(100 mg/kg)and indomethacin(5 mg/kg)has significant effect on CFA-induced AA rats,and its mechanism may be related to inhibiting CFA-induced oxidative stress and the activation of TLR4/NF-κB/NLRP3 signaling pathway,allevia-ting inflammatory reaction and correcting lysosome dysfunction.
Total triterpenoids from Chaenomeles speciosaIndomethacinAdjuvant arthritisInflammatory reactionLysosome functionTLR4/NF-κB/NLRP3 signaling pathway
万方数据
维普
