摘要
目的:基于AMP活化蛋白激酶(AMPK)/过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC1α)信号通路探讨膝痹宁II(XBN II)调控糖酵解并减轻膝骨关节炎(KOA)大鼠软骨损伤的作用机制。方法:将32只SD大鼠随机分为假手术(sham)组、KOA组、XBN II组和二甲双胍(Met)组,每组8只。通过前交叉韧带切断术建立KOA模型。造模后第14天开始,XBN II组大鼠每日予XBN II灌胃1次,Met组隔天予Met腹腔注射1次,共给药4周。采用HE和番红固绿染色评估软骨组织病理并进行Mankin和OARSI评分;乳酸检测试剂盒检测各组大鼠软骨组织和血清中乳酸含量;免疫组化检测软骨组织中磷酸化AMPK和PGC1α蛋白水平;RT-qPCR和Western blot检测糖酵解相关因子(葡萄糖转运蛋白1、己糖激酶2和乳酸脱氢酶A)、软骨合成和分解代谢相关因子(II型胶原、聚集蛋白聚糖、基质金属蛋白酶13和含血小板应答蛋白基序的解整联蛋白及金属蛋白酶5)及AMPK/PGC1α信号通路相关因子的mRNA和蛋白表达水平。结果:与sham组比较,KOA组大鼠软骨组织和血清中乳酸水平显著升高(P<0.05),软骨组织表面磨损,软骨层结构紊乱,基质淡染,Mankin和OARSI评分显著升高(P<0.05);软骨合成代谢因子及AMPK/PGC1α信号通路相关因子的mRNA和蛋白表达水平显著降低(P<0.05),软骨分解代谢相关因子及糖酵解相关因子的mRNA和蛋白表达水平显著升高(P<0.05)。与KOA组比较,XBN II组和Met组大鼠软骨组织形态有明显改善,Mankin和OARSI评分显著降低,且上述因子的mRNA和蛋白表达水平均显著逆转(P<0.05)。结论:XBN II可通过抑制糖酵解有效减轻KOA大鼠软骨损伤,其作用机制可能与AMPK/PGC1α信号通路的激活有关。
AIM:To investigate whether Xibining II(XBN II)attenuates cartilage damage in rats with knee osteoarthritis(KOA)by modulating glycolysis via the AMP-activated protein kinase(AMPK)/peroxisome proliferator-acti‑vated receptorγcoactivator 1α(PGC1α)signaling pathway.METHODS:Thirty-two SD rats were randomly divided into sham group,KOA group,XBN II group and metformin(AMPK activator)group,with 8 rats in each group.The rats in KOA group were subjected to the anterior cruciate ligament transection procedure to establish the KOA model.Starting from the 14th day after modeling,the rats in XBN II group received a daily dose of XBN II via gavage once a day,and those in metformin group were administered metformin via intraperitoneal injection once a day for 4 weeks.Subsequently,the histopathological changes of the cartilage were examined by HE and safranin O-fast green staining with matching Mankin and OARSI scores.The protein levels of phosphorylated AMPK(p-AMPK)and PGC1αin cartilage were quanti‑fied through immunohistochemistry.In addition,RT-qPCR and Western blot were conducted to measure the mRNA and protein expression levels of glycolysis-related factors,including glucose transporter 1,hexokinase 2 and lactate dehydroge‑nase A,biomarkers related to cartilage synthesis and catabolism,such as collagen type II,aggrecan,matrix metallopro‑teinase 13 and a disintegrin and metalloproteinase with thrombospondin motifs 5,and AMPK/PGC1αsignaling pathway-re‑lated indicators.RESULTS:Lactate levels in cartilage and serum were higher in KOA group compared with sham group(P<0.05).Similarly,the cartilage in KOA group exhibited significant surface abrasion and structural damage,with faintstained matrix and significantly higher Mankin and OARSI scores compared with sham group(P<0.05).Further analysis revealed significant decreases in the mRNA and protein expression levels of factors related to cartilage anabolism and AMPK/PGC1αsignaling pathway in KOA group compared with sham group(P<0.05).In contrast,there were marked in‑creases in the mRNA and protein expression levels of factors related to cartilage catabolism and glycolysis(P<0.05).No‑tably,XBN II and metformin treatments significantly improved the cartilage morphology,reduced Mankin and OARSI scores,and reversed the changes in mRNA and protein levels of the aforementioned indexes(P<0.05).CONCLU⁃SION:Treatment with XBN II can alleviate cartilage damage in KOA rats by inhibiting glycolysis,through a mechanism involving activation of the AMPK/PGC1αsignaling pathway.
作者
魏义保
马振源
刘德仁
胡恩睿
李晓辰
王培民
廖太阳
梅伟
WEI Yibao;MA Zhenyuan;LIU Deren;HU Enrui;LI Xiaochen;WANG Peimin;LIAO Taiyang;MEI Wei(Department of Orthopedics and Traumatology,Affiliated Hospital of Nanjing University of Chinese Medicine&Jiangsu Province Hospital of Chinese Medicine,Nanjing 210029,China;Department of Orthopedics and Traumatology,The Second Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210017,China)
出处
《中国病理生理杂志》
北大核心
2025年第8期1569-1577,共9页
Chinese Journal of Pathophysiology
基金
国家自然科学基金青年项目(No.82305276)
江苏省医学重点学科/实验室建设单位(No.JSDW202252)
江苏省中医院中医膝骨关节炎临床医学创新中心(No.Y2023zx05)
江苏省中医药科技发展计划青年人才项目(No.QN202105)。
作者简介
通讯作者:梅伟,E-mail:meiwei@njucm.edu.cn;通讯作者:廖太阳,E-mail:drtaiyang@126.com。
