摘要
目的探讨葛根舒筋方通过调控自噬治疗胶原诱导关节炎(CIA)大鼠的药效作用机制。方法将30只雄性Wistar大鼠随机分为正常组、模型组、甲氨蝶呤组(7.6 mg·kg^(-1))和葛根舒筋方低、中、高剂量组(4、8、16g·kg^(-1)),每组5只。除正常组外,各组大鼠均采用二次免疫法建立CIA大鼠模型。葛根舒筋方各剂量组和甲氨蝶呤组按相应剂量进行灌胃给药,其中葛根舒筋方各剂量组1次/d,甲氨蝶呤组1次/周,连续给药28d。实验结束后,测量大鼠体质量、足垫厚度以评估大鼠关节炎病情;采用HE及番红固绿染色法观察各组关节炎大鼠膝关节病理学变化;Elisa法检测各组大鼠血清中TNF-α、IL-6、IL-1β及IgG的表达情况;qPCR法检测各组关节炎大鼠踝关节中自噬相关基因Beclin-1、ATG5、ATG12、LC3B的表达水平。结果与正常组相比,模型组大鼠出现体质量下降、足爪红肿等症状,膝关节病理染色可见明显的滑膜增生、炎性浸润以及软骨损伤现象,血清炎症因子TNF-α、IL-6、IL-1β、IgG及踝关节组织中自噬相关基因Beclin-1、ATG5、ATG12、LC3B的表达水平较正常组显著升高(P<0.05);经药物干预后,各给药组大鼠体重较模型组均有升高,足爪红肿症状和膝关节中炎性浸润、滑膜增生现象显著缓解,骨与软骨组织结构更为完整;与模型组大鼠相比,血清炎症因子TNF-α、IL-6、IL-1β、IgG及踝关节中自噬相关基因Beclin-1、ATG5、ATG12、LC3B的表达水平显著降低,其中以葛根舒筋方高剂量组和甲氨蝶呤组的效果最为明显(P<0.05或P<0.01)。结论葛根舒筋方可通过调控自噬抑制CIA大鼠炎症反应,改善CIA大鼠病情。
Objective To investigate the pharmacological mechanism of Gegen(Radix Puerariae Lobatae)Shujin Formula(葛根舒筋方,GGSJF)in treating collagen-induced arthritis(CIA)rats by regulating autophagy.Methods Thirty male Wistar rats were randomly divided into the normal group,model group,methotrexate group(7.6 mg·kg^(-1)),and low/medium/high-dose GGSJF groups(4,8,16 g·kg^(-1)),with 5 rats in each group.Except for the normal group,CIA models were established in all other groups by a secondary immunization method.The GGSJF groups and the methotrexate group were administered via intragastric gavage at their respective doses.Specifically,each GGSJF group received daily administration,while the methotrexate group was administered weekly,with continuous administration maintained for 28 consecutive days.After the experiment,body weight and paw thickness of rats were measured to evaluate the arthritis severity.Hematoxylin-eosin(HE)and safranin O-fast green staining were used to observe the pathological changes in knee joints.ELISA was used to detect the expressions of TNF-α,IL-6,IL-1β,and IgG in serum.The expression levels of autophagy-related genes of Beclin-1,ATG5,ATG12,LC3B in ankle joints were measured by qPCR.Results Compared with the normal group,the model group showed decreased body weight,and red and swollen paws.Obvious synovial hyperplasia,inflammatory infiltration,and cartilage damage could be observed through pathological staining of the knee joint.The expression levels of serum inflammatory factors TNF-α,IL-6,IL-1β,IgG,and autophagy-related genes Beclin-1,ATG5,ATG12,and LC3B in the ankle joint tissues were significantly increased compared with the normal group(P<0.05).After drug intervention,all treatment groups exhibited symptoms such as decreased body weight,and red and swollen paws,alleviated inflammatory infiltration in the knee joint and synovial hyperplasia,and improved bone and cartilage structure.The expression levels of serum inflammatory factors TNF-α,IL-6,IL-1β,IgG,and autophagy-related gene Beclin-1,ATG5,ATG12,and LC3B in the ankle joint were significantly reduced compared to the model group,with the high-dose GGSJF and methotrexate groups showing the most pronounced effects(P<0.05 or P<0.01).Conclusion GGSJF may inhibit inflammatory responses and ameliorate CIA symptoms in rats by regulating autophagy.
作者
李宜桓
庞彩霞
谭淑慧
阙嘉露
柳璇
周鹏
梁奇
刘翠玲
LI Yihuan;PANG Caixia;TAN Shuhui;QUE Jialu;LIU Xuan;ZHOU Peng;LIANG Qi;LIU Cuiling(School of the Seventh Clinical Medicine of Guangzhou University of Chinese Medicine,Shenzhen Bao’an Traditional Chinese Medicine Hospital,Shenzhen 518133,China)
出处
《时珍国医国药》
北大核心
2025年第15期2847-2853,共7页
JOURNAL OF LI-SHIZHEN TRADITIONAL CHINESE MEDICINE
基金
广东省基础与应用基础研究基金项目(2019A1515111192)
深圳市宝安区科技创新局项目(2019JD316)
深圳市“医疗卫生三名工程”项目(SZZYSM202206005)。
关键词
葛根舒筋方
胶原诱导关节炎
类风湿关节炎
自噬
Gegen Shujin Formula(葛根舒筋方,GGSJF)
Collagen-induced arthritis(CIA)
Rheumatoid arthritis
Autophagy
