虫草素调节Wnt/β-catenin信号通路对肩袖撕裂损伤后脂肪浸润的影响  

Cordycepin Inhibits Fat Infiltration after Rotator Cuff Tear Injury by Regulating Wnt/β-catenin Signaling Pathway

作　　者：谢求恩[1,2] 梁邓文 吴韶 郝旭辉 梁利光 简帮祥 董俊宏 杨雷 XIE Qiu'en;LIANG Dengwen;WU Shao;HAO Xuhui;LIANG Liguang;JIAN Bangxiang;DONG Junhong;YANG Lei(The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;The First Clinical College of Traditional Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410007,China)

机构地区：[1]湖南中医药大学第一附属医院,长沙410007 [2]湖南中医药大学第一中医临床学院,长沙410007

出　　处：《中国实验方剂学杂志》2025年第3期98-106,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：湖南省教育厅科学研究项目(22A0244);湖南省卫生健康委科研课题(202204074105);国家重点实验室培育基地开放基金项目(24PTKF1011)。

摘　　要：目的:研究虫草素通过调控Wnt/β-连环蛋白(β-catenin)信号通路抑制大鼠肩袖损伤后脂肪浸润的作用及其机制,为临床治疗肩袖损伤提供理论依据。方法:SPF级雌性SD大鼠50只,随机选取10只作为正常组,采用冈上肌肌腱和肩胛上神经压迫建立肩袖损伤修复模型,将手术成功的大鼠按照随机区组设计分为模型组、虫草素低、中、高剂量组(20、40、80 mg·kg^(-1))。治疗6周后,采用步态分析评估大鼠肢体功能;油红O染色和马松(Masson)染色观察肌肉组织病理变化情况;酶联免疫吸附测定法(ELISA)检测血清中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量;免疫组化(IHC)检测脂肪形成标记物氧化物酶体增殖物激活受体γ(PPARγ)和CCAAT增强子结合蛋白α(C/EBPα)蛋白的表达,实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白免疫印迹法(Western blot)检测Wnt3a、Wnt10b、β-catenin mRNA和蛋白表达情况。结果:与正常组比较,模型组大鼠步幅长和爪印面积显著下降(P<0.01),湿肌质量减少比率明显升高、肌纤维横截面积明显下降(P<0.05),脂肪浸润面积比率下降、胶原纤维面积比率显著下降(P<0.01),IL-1β、IL-6和TNF-α含量明显上升(P<0.05),PPARγ、C/EBPα蛋白表达均显著升高(P<0.01),Wnt3a、Wnt10b、β-catenin mRNA和蛋白表达均明显降低(P<0.05,P<0.01);与模型组比较,虫草素低、中、高剂量组大鼠步幅长和爪印面积均显著升高(P<0.01),湿肌质量减少比率明显降低、肌纤维横截面积明显升高(P<0.05),脂肪浸润面积比率和胶原纤维面积比率明显升高(P<0.05,P<0.01),IL-1β、IL-6、TNF-α含量明显下降(P<0.05,P<0.01),PPARγ、C/EBPα蛋白表达均显著降低(P<0.01),Wnt3a、Wnt10b、β-catenin mRNA和蛋白表达均明显升高(P<0.05,P<0.01)。结论:虫草素能显著改善大鼠肢体功能,减轻大鼠肩袖肌肉萎缩、脂肪浸润及纤维化程度,并能抑制炎症反应,其�Objective:To investigate the effect and mechanism of cordycepin in inhibiting fat infiltration after rotator cuff injuries in rats by regulating the Wnt/β-catenin signaling pathway,providing a theoretical basis for clinical treatment of rotator cuff injuries.Methods:Fifty SPF-grade female SD rats were used in this study,with 10 randomly selected as the blank group.A rotator cuff injury repair model was established by supraspinatus tendon and suprascapular nerve compression.The successfully modeled rats were randomized into model and low-dose(20 mg·kg^(-1)),medium-dose(40 mg·kg^(-1)),and high-dose(80 mg·kg^(-1))cordycepin groups.After 6 weeks of treatment,the gait analysis was performed to assess the limb function in rats.Oil red O staining and Masson staining were employed to observe pathological changes in the muscle tissue.Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in the serum.Immunohistochemistry(IHC)was employed to detect the expression of peroxisome proliferator-activated receptorγ(PPARγ)and CCAAT/enhancer-binding proteinα(C/EBPα),which are markers of adipogenesis.Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)and Western blot were employed to determine the mRNA and protein levels,respectively,of Wnt3a,Wnt10b,andβ-catenin.Results:Compared with the blank group,the model group showed decreases in stride length and paw print area(P<0.01),an increase in ratio of wet muscle mass reduction and a decrease in muscle fiber cross-sectional area(P<0.05),and decreased ratios of fat infiltration area and collagen fiber area(P<0.01).Additionally,the model group showed elevated levels of IL-1β,IL-6,and TNF-α(P<0.05),up-regulated protein levels of PPARγand C/EBPα(P<0.01),and down-regulated mRNA and protein levels of Wnt3a,Wnt10b,andβ-catenin(P<0.05,P<0.01).Compared with the model group,the low-,medium-,and high-dose cordycepin groups showed increases in stride length and

关 键 词：虫草素 肩袖损伤 Wnt/β-连环蛋白(β-catenin) 脂肪浸润 

分 类 号：R285[医药卫生—中药学] R243[医药卫生—中医学] R264