摘要
目的 探讨桂枝芍药知母汤(GSZD)的抗类风湿关节炎(RA)作用,并初步探讨其可能的作用机制。方法 72只雌性Wistar大鼠随机分为对照组,模型组,阳性药组,GSZD低、中、高剂量组,除对照组外,其余各组均采用胶原诱导法建立胶原诱导性关节炎(CIA)大鼠模型。建模成功后给予相应治疗方案,GSZD低、中、高剂量组[GSZD 0.8、1.6、3.2 g/(kg·d)]分别每日灌胃1次;阳性药组[甲氨蝶呤(0.75 mg/(kg·d)]每3日灌胃1次;对照组和模型组大鼠灌胃等体积生理盐水,每日1次,治疗周期为14 d。记录大鼠关节炎指数(AI)评分、足肿胀度和免疫器官指数的变化情况;通过Micro-CT和HE染色评估踝关节组织滑膜增生和关节软骨损伤情况;酶联免疫吸附实验(ELISA)检测大鼠血清中炎症因子水平变化;免疫组化和Western blot检测软骨中自噬相关蛋白的表达情况。结果 与对照组相比,模型组大鼠出现滑膜增生、血管翳形成、软骨侵蚀和骨破坏等病理性改变,AI评分、足肿胀度、免疫器官指数,血清肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6水平,踝关节组织和软骨中p62、p-mTOR蛋白表达显著升高;血清IL-10、踝关节组织和软骨中LC3BⅡ/LC3BⅠ、p-AMPK、p-ULK-1蛋白表达水平显著降低。与模型组相比,GSZD各剂量组和阳性药组大鼠病理性关节损伤明显改善;AI评分,足肿胀度,免疫器官指数,血清中TNF-α、IL-1β、IL-6水平,踝关节组织和软骨中p62、p-mTOR蛋白表达显著降低;血清IL-10,踝关节组织和软骨中LC3BⅡ/LC3BⅠ、p-AMPK、p-ULK-1蛋白表达水平显著升高。结论 GSZD能够抑制CIA大鼠的炎症症状和骨破坏,其机制可能与调节AMPK/mTOR/ULK-1信号通路促进自噬有关。
Objective To determine the anti-rheumatoid arthritis(RA)effect of Guizhi Shaoyao Zhimu decoction(GSZD)and its potential mechanism.Methods Totally 72 female Wistar rats were randomly allocated into six groups:a control group,a model group,a positive(methotrexate)group,and GSZD low,medium,and high doses groups.Except for the control group,collagen-induced arthritis(CIA)model was induced in the remaining groups with collagen.Upon successful establishment of the model,the treatment protocols were implemented as follows:the GSZD low,medium,and high dose groups received oral administrations of 0.8,1.6,and 3.2 g/(kg·d),respectively,once daily while the positive group received an oral dose of 0.75 mg/(kg·d)methotrexate every three days.The control and model groups were given equivalent volumes of normal saline via gavage once daily over 14 days.We monitored changes in the arthritis index score,foot swelling,and immune organ index in the rats.Synovial hyperplasia and articular cartilage damage in the ankle joint tissue were assessed with Micro-CT and HE staining.ELISA was used to measure changes in inflammatory cytokine levels in the rat serum,while immunohistochemistry and Western blot were used to determine the expression of autophagy-related proteins in the cartilage.Results Compared with the control group,the model group displayed notable pathological alterations,including synovial hyperplasia,vascular opacity formation,cartilage erosion,and bone destruction.There was a significant increase in the arthritis index score,foot swelling degree,immune organ index,and serum inflammatory factors such as levels of tumor necrosis factor(TNF)-α,Interleukin(IL)-1β,and IL-6,as well as the expression of p62 and p-mTOR proteins in the ankle joint tissue and cartilage.Conversely,the expression levels of serum IL-10,LC3BⅡ/LC3BⅠ,p-AMPK,and p-ULK-1 proteins in the ankle joint tissue and cartilage were significantly decreased.In contrast,rats in the GSZD dose groups and the positive drug group demonstrated significant improvements in pathological joint injuries as compared with those of the model group.There was a marked reduction in arthritis index scores,foot swelling degree,immune organ index,serum levels of inflammatory factors TNF-α,IL-1β,and IL-6 levels,and the expression of p62 and p-mTOR proteins in the ankle joint tissue and cartilage.There was a significant increase in the expression levels of serum IL-10,LC3BⅡ/LC3BⅠ,p-AMPK,and p-ULK-1 proteins in the ankle joint tissue and cartilage.Conclusion GSZD can mitigate inflammatory symptoms and prevent bone destruction in collagen-induced arthritis rats,which may be potentially mediated through the modulation of the AMPK/mTOR/ULK-1 signaling pathway to promote autophagy.
作者
华重钧
周政署
梁霄
王锐
姜德友
HUA Chong-jun;ZHOU Zheng-shu;LIANG Xiao;WANG Rui;JIANG De-you(Basic Medical College,Heilongjiang University of Traditional Chinese Medicine,Harbin 150040;College of Pharmacy,Heilongjiang University of Traditional Chinese Medicine,Harbin 150040)
出处
《中南药学》
2025年第5期1221-1228,共8页
Central South Pharmacy
基金
国家自然科学基金资助项目(No.81603418)
国家中医药管理局龙江医学流派传承工作室建设项目(No.LPGZS2012-14)。
作者简介
华重钧,女,硕士研究生,主要从事经方治疗疑难病的临床与基础研究,email:3489975689@qq.com;通信作者:姜德友,男,教授,博士研究生导师,主要从事经方治疗疑难病的临床与基础研究,email:jiangdeyou@126.com。
