穿山龙薯蓣皂苷通过调控自噬治疗胶原诱导性关节炎小鼠模型的机制  

Mechanism research of pangolin diosgenin in treating collagen-induced arthritis mouse model by regulating autophagy

作　　者：江涛 由涌 夏东帅 薄思涵 王永为 高亚贤 宋鸿儒 JIANG Tao;YOU Yong;XIA Dong-shuai;BO Si-han;WANG Yong-wei;GAO Ya-xian;SONG Hong-ru(Dept of Immunology,Basic Medical Institute,Chengde Medical College,Chengde Hebei 067000,China;Dept of Anatomy,Basic Medical Institute,Chengde Medical College,Chengde Hebei 067000,China;Heibei North University,Zhangjiakou Hebei 075000,China)

机构地区：[1]承德医学院基础医学院免疫学教研室,河北承德067000 [2]承德医学院基础医学院解剖学教研室,河北承德067000 [3]河北北方学院医学检验学院免疫学教研室,河北张家口075000

出　　处：《中国药理学通报》2024年第4期645-651,共7页Chinese Pharmacological Bulletin

基　　金：河北省自然科学基金中医药联合基金培育项目(No H2023406057);河北省自然科学基金精准医学联合基金细化项目(No H2021406066);河北省高等学校科学技术研究项目(No ZD2021003)。

摘　　要：目的探讨穿山龙薯蓣皂苷通过自噬治疗胶原诱导性关节炎(collagen-induced arthritis,CIA)模型小鼠的作用机制。方法通过鸡Ⅱ型胶原乳剂建立CIA模型小鼠,初次免疫DBA1小鼠(0.1 mL),第21天后再次给予相同剂量刺激。加强免疫后将胶原免疫的50只小鼠随机分组为CIA模型组、雷公藤组和穿山龙薯蓣皂苷低、中、高剂量组,另有10只小鼠注射等量生理盐水作为对照组。在对小鼠进行连续14天灌胃治疗并观察各组小鼠足爪的炎症程度,取下各组小鼠脾、腹股沟淋巴结和踝关节、膝关节,对踝关节、膝关节进行HE染色,检测各组淋巴结、脾组织中mRNA和蛋白质水平的相关自噬指标。结果与CIA模型组相比,穿山龙薯蓣皂苷高剂量组关节炎指数(AI)评分降低(P<0.05);雷公藤组及穿山龙薯蓣皂苷组足爪厚度差值减小(P<0.01)。与对照组相比,CIA模型组体质量明显下降(P<0.01);小鼠膝关节滑膜和足爪踝关节HE染色结果显示CIA模型组对比对照组出现免疫细胞浸润、骨质侵蚀、软骨破坏和滑膜增生的症状;雷公藤组与穿山龙薯蓣皂苷中、高剂量组对CIA模型有不同程度的治疗作用,可以减轻和预防症状。与CIA模型组相比,雷公藤组和穿山龙薯蓣皂苷治疗组中LC3、Beclin-1的mRNA水平和蛋白水平的表达升高,p62的mRNA水平和蛋白水平的表达降低。结论穿山龙薯蓣皂苷可能通过抑制自噬对CIA模型小鼠起到治疗作用。Aim To explore the mechanism of action of pangolin diosgenin in treating collagen-induced arthritis(CIA)model mice through autophagy.Methods CIA mouse model was established using chicken type II collagen emulsion.DBA1 mice were initially immunized(0.1 mL)and stimulated again with the same dose after 21 days.After boosting the immunization,50 mice immunized with collagen were randomly divided into the CIA model group,tripterygium wilfordii group and pangolin diosgenin low,medium and high dose groups,and another 10 mice were injected with the same amount of normal saline as the control group.After 14 consecutive days of intragastric treatment,the spleen,inguinal lymph nodes and knee joints of CIA mice were removed.The degree of inflammation in the paws of mice in each group was observed.HE staining was performed on ankle joints and knee joints,and related autophagy indicators at mRNA and protein levels in lymph nodes and spleen tissue of each group were detected.Results Compared with the CIA model group,the arthritis index(AI)score of the high-dose pangolin diosgenin group was reduced(P<0.05);the difference in paw thickness between the tripterygium wilfordii group and the pangolin diosgenin group was reduced(P<0.01).Compared with the control group,the weight of the CIA model group decreased significantly(P<0.01).HE staining results of mouse knee joints synovium and paw ankle joints showed that the CIA model group had symptoms of immune cell infiltration,bone erosion,cartilage destruction,and synovial hyperplasia compared with the control group.The tripterygium wilfordii group and the medium-and high-dose groups of pangolin diosgenin had different degrees of therapeutic effect on the CIA model,and could reduce and prevent symptoms.Compared with the CIA model group,the mRNA and protein levels of LC3 and Beclin-1 increased in the tripterygium wilfordii group and the pangolin diosgenin-treated group,while the mRNA and protein levels of p62 decreased.Conclusion Pangolin diosgenin may have a therapeutic effect on CIA mod

关 键 词：穿山龙薯蓣皂苷 自噬 类风湿性关节炎 胶原诱导性关节炎 P62 LC3 BECLIN-1 

分 类 号：R-332[医药卫生] R284.1R341.26R392R593.22R977.6