基于PI3K/AKT/FOXO1信号通路探究三强方对膝骨关节炎大鼠股四头肌萎缩的影响OA
Exploring the effect of Sanqiang formula on quadriceps muscle atrophy in knee osteoarthritis in rats through PI3K/AKT/FOXO1 signaling pathway
目的 通过三强方干预膝骨关节炎(KOA)大鼠,观察该方对KOA大鼠股四头肌的影响,进一步探究PI3K/AKT/FOXO1信号通路中增殖凋亡相关蛋白及mRNA的变化,初步探索三强方对失稳性膝骨关节炎股四头肌萎缩的作用机制,为该方临床应用和开发提供实验依据.方法 采用大鼠右后膝关节前交叉韧带离断法构建KOA大鼠模型,假手术组仅切开皮肤,完整保留前交叉韧带,将造模成功的大鼠分为模型组、塞来昔布组、三强低、中、高剂量组,每组10只.假手术组和模型组给予0.9％氯化钠溶液灌胃,其余各组给予相应剂量的药物灌胃,给药前后分别检测大鼠行为学.各组大鼠给药结束后,检测大鼠膝关节被动活动度;测定大鼠股四头肌重量、大鼠血清白细胞介素-1β(IL-1β)及肿瘤坏死因子-α(TNF-α)水平;采用HE染色观察各组SD大鼠股四头肌组织损伤情况;采用Masson染色观察股四头肌纤维化改变;采用HE染色观察软骨组织变化;RT-qPCR检测各组骨骼肌组织Atrogin-1和MuRF1mRNA的表达;Western blot法检测各组骨骼肌组织PI3K、p-PI3K、AKT、p-AKT、FOXO1、p-FOXO1、Atrogin-1和MuRF1蛋白的表达.结果 三强方可改善大鼠行为学变化(P＜0.05,P＜0.001);使被动活动度显著提升(P＜0.001);提升股四头肌重量(P＜0.05);降低血清中IL-1β及TNF-α含量(P＜0.01,P＜0.001);中、高剂量组提升肌细胞面积(P＜0.001);显著降低各组间质结缔组织的含量,降低胶原容积分数(P＜0.001);改善关节软骨的病理形态学改变;升高股四头肌中p-PI3K/PI3K、p-AKT/AKT、p-FOXO1/FOXO1 的含量(P＜0.05,P＜0.01,P＜0.001);抑制股四头肌中 Atrogin-1、MuRF1 蛋白和mRNA的表达(P＜0.01,P＜0.001).结论 三强方可促进KOA大鼠股四头肌PI3K/AKT/FOXO1信号通路活化并抑制Atrogin-1和MuRF1的表达,改善股四头肌的萎缩.
Objective This study aimed to preliminarily investigate the mechanism by which Sanqiang formula acts on quadriceps muscle atrophy in unstable knee osteoarthritis(KOA),and to provide experimental evidence for its clinical application and development.Methods The KOA rat model was established by transection of the anterior cruciate ligament(ACL)of the right posterior knee joint.In the Sham operation group,only the skin was incised,and the ACL was completely preserved.The successfully modeled rats were divided into the Model group,celecoxib group,and low-,medium-,and high-dose Sanqiang formula groups,with ten rats in each group.The Sham operation and model groups were administered normal saline by gavage;the other groups were administered the corresponding doses of the drug by gavage.Behavioral tests were conducted before and after drug administration.After the end of administration,the passive range of motion of the rat knee joint was measured;the quadriceps femoris muscle weight,and serum interleukin(IL)-1β and tumor necrosis factor(TNF)-α levels were determined.Hematoxylin-eosin staining was used to observe quadriceps muscle tissue damage in SD rats of each group,and Masson staining was used to observe quadriceps muscle fibrosis changes.RT-qPCR was used to detect the expression of Atrogin-1 and MuRF1 mRNA in skeletal muscle tissues of each group;Western blot was used to detect the expression of PI3K,p-PI3K,AKT,p-AKT,FOXO1,p-FOXO1,Atrogin-1,and MuRF1 proteins in skeletal muscle tissues of each group.Results Sanqiang formula improved behavioral changes in rats(P＜0.05,P＜0.001),significantly increased passive range of motion(P＜0.001),and increased quadriceps muscle weight(P＜0.05).It decreased serum IL-1β and TNF-α(P＜0.01,P＜0.001),increased muscle cell area in the medium-and high-dose groups(P＜0.001),significantly reduced interstitial connective tissue among groups,and reduced collagen volume fraction(P＜0.001).Treatment improved pathological morphological changes in articular cartilage,increased p-PI3K/PI3K,p-AKT/AKT,and p-FOXO1/FOXO1 values in quadriceps muscle(P＜0.05,P＜0.01,P＜0.001),and inhibited Atrogin-1 and MuRF1 protein and mRNA expression in quadriceps muscle(P＜0.01,P＜0.001).Conclusions Sanqiang formula promoted the activation of PI3K/AKT/FOXO1 signaling in the quadriceps muscle of KOA rats,inhibited the expression of Atrogin-1 and MuRF1,and improved quadriceps muscle atrophy.
王翀;王静雅;闫康;虞跃跃;李继安
华北理工大学中医学院,河北唐山 063210华北理工大学中医学院,河北唐山 063210||广安门医院保定医院,河北保定 071000河北大学中医学院,河北保定 071000华北理工大学中医学院,河北唐山 063210华北理工大学中医学院,河北唐山 063210||河北省国际联合研究中心,河北唐山 063210
医药卫生
三强方膝骨关节炎股四头肌PI3K/AKT/FOXO1信号通路
Sanqiang formulaknee osteoarthritisquadriceps musclePI3K/AKT/FOXO1 signaling pathway
《中国比较医学杂志》 2026 (7)
40-52,13
河北省国际合作专项(19392507D)科技部对发展中国家常规性科技援助项目(KY201904005).
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