摘要
旨在以Hippo-YAP/TAZ通路为切入点,揭示并验证白果内酯(bilobalide,BB)改善前十字韧带切断(anterior cruciate ligament transection,ACLT)构建的大鼠骨关节炎(Osteoarthritis,OA)模型软骨退变的作用机制。选取45只雄性Sprague-Dawley(SD)大鼠随机分为对照组、模型组(OA组)和药物干预组(BB组)。除对照组外,采用ACLT方法构建大鼠OA模型。BB组大鼠每天灌胃10 mg·kg^(-1)BB,连续给药6周后取膝关节和软骨组织样本。手术造模后每周对每组大鼠进行一次关节肿胀检测。通过大体观察及Pelletier评分和组织学观察及OARSI评分评估大鼠软骨退变程度,Micro-CT观测软骨下骨损伤程度,免疫印迹法检测Hippo-YAP/TAZ通路相关蛋白MST1、LATS1、YAP和TAZ水平,以及软骨细胞外基质(cartilage extracellular matrix,ECM)降解蛋白MMP-3变化。计算机分子对接预测BB与潜在靶点的结合力。结果显示,在ACLT诱导的大鼠OA模型中,BB能够抑制大鼠关节肿胀,抑制软骨病理特征并极显著降低OARSI评分(P<0.01),以及软骨下骨骨小梁结构的破坏,改善OA软骨退变。在机制上,BB极显著增加大鼠软骨Hippo-YAP/TAZ通路中MST1和LATS1水平(P<0.01),抑制核内YAP和TAZ表达(P<0.01),并且降低MMP-3表达(P<0.01),发挥抗ECM降解作用。此外,分子对接显示BB与YAP和TAZ均具有较好的亲和性(结合能<-20.92 kJ·mol^(-1))。结果提示,BB通过Hippo-YAP/TAZ通路抑制大鼠软骨ECM降解,缓解关节肿胀并抑制软骨退变,发挥改善OA的作用。
The aim of this study was to reveal and validate the improvement effect of bilobalide(BB)on cartilage degeneration in a rat osteoarthritis(OA)model induced by anterior cruciate ligament transection(ACLT)via the Hippo-YAP/TAZ pathway.Forty-five male Sprague-Dawley(SD)rats were randomly divided into the control group,model group(OA group)and drug intervention group(BB group).In addition to the control group,the rat OA model was constructed by ACLT method.The rats in the BB group were given 10 mg·kg^(-1)BB daily for 6 weeks,after that,knee joint and cartilage tissue samples were taken for consequtive examination.After the operation,the rats in each group were examined once a week for joint swelling.The degree of cartilage degeneration of rats was evaluated by gross observation,Pelletier score,histological observation and the OARSI score.The degree of subchondral bone injury was observed by Micro-CT,and the levels of Hippo-YAP/TAZ pathway related proteins MST1,LATS1,YAP and TAZ,as well as the cartilage extracellular matrix(ECM)degradation protein MMP-3 were detected by Western blot.Computer molecular docking was used to predict the binding affinity of BB to potential targets.The results showed that in an ACLT-induced OA model,BB could inhibit joint swelling,alleviate cartilage pathological features and highly significantly reduce OARSI score(P<0.01).It also mitigated damage to the subchondral trabecular structure and ameliorated OA-related cartilage degeneration.In terms of mechanism,BB highly significantly increased the levels of MST1 and LATS1 in Hippo-YAP/TAZ pathway in rat cartilage(P<0.01),inhibited the expression of YAP and TAZ in the nucleus(P<0.01),and decreased the expression of MMP-3(P<0.01),thus playing an anti-ECM degradation role.In addition,molecular docking showed that BB had good affinity with YAP and TAZ(binding energy<-20.92 kJ·mol-1).The results suggested that BB inhibited the degradation of cartilage ECM,alleviated joint swelling and inhibited cartilage degeneration through Hippo-YAP/TAZ pathway,and played a role in improving OA.
作者
裘宗盛
李淑欣
戚晶晶
郭晓艳
马玉辉
张志恒
王瑞龙
王海阳
李亚楠
马天文
QIU Zongsheng;LI Shuxin;QI Jingjing;GUO Xiaoyan;MA Yuhui;ZHANG Zhiheng;WANG Ruilong;WANG Haiyang;LI Yanan;MA Tianwen(Heilongjiang Key Laboratory for Laboratory Animals and Comparative Medicine,College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China;Xinjiang Zhaosu County Xiyu Horse Industry Co.,LTD,Zhaosu 835500,China;College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010000,China)
出处
《畜牧兽医学报》
北大核心
2025年第12期6422-6430,共9页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金项目(32402967)
中国博士后科学基金项目(2025M774143)
黑龙江省自然科学基金项目(YQ2023C015)
黑龙江省博士后资助项目(LBH-Z23084)。
