木瓜总三萜和吲哚美辛联用对弗氏完全佐剂诱导大鼠类风湿性关节炎的治疗作用研究  

Therapeutic Effect of Total Triterpenoids of Chaenomeles speciosa Combined with Indomethacin on Rheumatoid Arthritis Induced by Freund's Complete Adjuvant in Rats

作　　者：贺君宇 覃诗颖 郑婧琪 罗识 熊致远 余清泉 HE Junyu;QIN Shiyin;ZHENG Jingqi;LUO Shi;XIONG Zhiyuan;YU Qingquan(Three Gorges University,College of Basic Medical Sciences,Yichang 443002;Nantong University Medical School,Nantong 226001;College of Biological and Pharmaceutical Sciences of China Three Gorges University,Yichang 443002)

机构地区：[1]三峡大学基础医学院,宜昌443002 [2]南通大学医学院,南通226001 [3]三峡大学生物与制药学院,宜昌443002

出　　处：《中药药理与临床》2024年第8期42-50,共9页Pharmacology and Clinics of Chinese Materia Medica

基　　金：湖北省宜昌市科学技术局自然科学基金项目(编号:A21-2-044);湖北省科技厅自然科学基金项目(编号:2022CFB357、2022CFB427);湖北省卫生健康委员会中医药重点项目(编号:ZY2023Z015)。

摘　　要：目的:观察木瓜总三萜和吲哚美辛联用对佐剂性关节炎大鼠的治疗作用,并探寻其可能作用机制。方法:将大鼠随机分为正常对照组、模型对照组、吲哚美辛5 mg/kg组、木瓜总三萜100 mg/kg组、木瓜总三萜100 mg/kg+吲哚美辛5 mg/kg联用组。除正常对照组外,各组大鼠右后脚底皮下注射弗氏完全佐剂(CFA)。14 d后,灌胃给予相应药物,1次/d,连续28 d,计算足肿胀度及关节炎指数评分。末次给药次日,取血和踝关节滑膜组织,检测血清中活性氧(ROS)、乳酸脱氢酶(LDH)和内源性抗氧化酶活性,踝关节中炎症因子含量、滑膜组织和溶酶体中组织蛋白酶B和D、β-半乳糖苷酶和β-葡萄糖醛酸酶活性及形态学分析;实时定量PCR和Western blot法检测踝关节组织中Toll样受体4/核因子κB/NOD样受体家族3(TLR4/NF-κB/NLRP3)通路相关的mRNA和蛋白表达。结果:与正常对照组比较,模型对照组大鼠足肿胀度、关节炎指数评分显著增加(P<0.01),软骨破坏和炎性浸润出现,血清中ROS、丙二醛(MDA)、髓过氧化物酶(MPO)、LDH和踝关节组织中环氧酶(COX-1)、COX-2、前列腺素E2(PGE2)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、IL-6、IL-18及踝关节组织蛋白酶B、D、β-半乳糖苷酶、β-葡萄糖醛酸酶和溶酶体细胞器胞质中β-葡萄糖醛酸酶、组织蛋白酶D含量显著升高(P<0.01),血清中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)和踝关节组织中IL-4、IL-10含量及细胞器溶酶体中组织蛋白酶D、β-葡萄糖醛酸酶活力或含量显著降低(P<0.01),踝关节组织中Tlr4、髓样分化因子(Myd88)、Nlrp3、凋亡相关的斑点样蛋白(Asc)、半胱氨酸天冬氨酸蛋白酶-1(Caspase1)、Cox1、Cox2、Pge2mRNA及磷酸化IκB激酶-α(p-IκBα)、磷酸化KB抑制蛋白激酶β(p-IKKβ)、硫氧还蛋白互作蛋白(TXNIP)、细胞有丝分裂相关酶(NEK7)、NLRP3、ASC、Pro-caspase-1、Caspase-1、�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 triterpenoids of C.speciosa 100 mg/kg and indomethacin 5 mg/kg).Except for the normal control group,rats in each group were subcutaneously injected 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 antioxidant enzymes in serum,inflammatory factors in synovial tissues,the activities of cathepsins B and D,β-glucuronidase,β-galactosidase in synovial 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 destruction and inflammatory infiltration,and up-regulated ROS,lipid peroxidation(MDA),myeloperoxidase(MPO),LDH in serum,elevated cyclooxygenases-2(COX-1),COX-2,prostaglandin E_2(PGE_2),IL-1β,TNF-α,IL-6,IL-18,cathepsins B and D,β-galactosidase,β-glucuronidase in synovial tissues,and enhanced activities ofβ-galactosidase and cathepsin D in the cytoplasm of lysosomal organelles(P<0.01).Additionally,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 activit

关 键 词：木瓜总三萜 吲哚美辛 佐剂性关节炎 炎症反应 溶酶体功能 Toll样受体4/核因子κB/NOD样受体家族3信号通路 

分 类 号：R285.5[医药卫生—中药学]