基于GLP-1/GLP-1R轴探讨黄芪治疗膝骨关节炎的机制研究  

Mechanism of Astragali Radix in treatment of knee osteoarthritis based on GLP-1/GLP-1R axis

作　　者：陈佳 王建国 王桂宇 吴婧若 岳进茹 刘琪 刘静淑 CHEN Jia;WANG Jian-guo;WANG Gui-yu;WU Jing-ruo;YUE Jin-ru;LIU Qi;LIU Jing-shu(College of Basic Medical Sciences,Shanxi University of Chinese Medicine,Jinzhong 030600,China;Shanxi Provincial Integrated Traditional Chinese Medicine and Western Medicine Hospital,Taiyuan 030000,China)

机构地区：[1]山西中医药大学基础医学院,山西晋中030600 [2]山西省中西医结合医院,山西太原030000

出　　处：《中国中药杂志》2024年第22期6190-6197,共8页China Journal of Chinese Materia Medica

基　　金：山西省自然科学基金面上项目(202103021224298);山西省中医药科研课题(2023ZYYB049);山西省高等学校科技创新项目(2022山西教育厅项目)(2022L352);山西中医药大学创新能力培育项目(2023-PY-TH-01);研究生精品教材项目(2023JC20)。

摘　　要：观察不同剂量黄芪对膝骨关节炎(KOA)大鼠血清胰高血糖素(GLP-1)和软骨组织中胰高血糖素受体(GLP-1R)的表达,探究黄芪调控GLP-1/GLP-1R信号轴对膝骨关节炎炎症和凋亡的影响,及其缓解KOA的作用机制。48只雄性SD大鼠随机分6个组:空白组,模型组,黄芪低、中、高剂量组(3.125、6.25、12.5 g·kg^(-1)),硫酸氨基葡萄糖组(0.1 g·kg^(-1))。除空白组外,其余大鼠均采用碘乙酸钠(MIA)膝关节腔注射构建膝KOA模型。造模成功后,连续治疗5周,酶联免疫吸附法(ELISA)检测大鼠血清GLP-1、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)水平;病理学检测观察膝关节软骨病理变化;qRT-PCR检测大鼠膝关节软骨TNF-α、基质金属蛋白酶13(MMP13)mRNA水平,蛋白免疫印迹法(Western blot)检测大鼠膝关节GLP-1R、MMP13、半胱氨酸蛋白酶8(caspase-8)蛋白表达水平;免疫组化检测膝关节GLP-1R、MMP13表达;TUNEL染色观察膝关节软骨细胞凋亡情况。以上实验结果显示黄芪可能通过GLP-1/GLP-1R轴提高血清GLP-1水平,降低血清中的TNF-α、IL-1β水平,降低TNF-α、MMP13 mRNA的相对表达,激活GLP-1R,降低软骨中MMP13、caspace-8蛋白的表达,调控其相关信号通路改善炎症、调控凋亡来达到保护软骨,改善膝骨关节炎的作用。This study aims to observe the effects of different doses of Astragali Radix on the expression of glucagon(GLP-1)in se-rum and glucagon receptor(GLP-1R)in cartilage tissue in rats with knee osteoarthritis(KOA),explore the effect of Astragali Radix on the inflammation and apoptosis of KOA by regulating GLP-1/GLP-1R signaling axis,and investigate the mechanism of its action in alleviating KOA.Forty-eight male SD rats were randomly divided into six groups:blank group,model group,low-,medium-,and high-dose Astragali Radix groups(3.125,6.25,and 12.5 g·kg^(-1)),and glucosamine sulfate group(0.1 g·kg^(-1)).Except for the blank group,rats in other groups were injected with sodium iodoacetate(MIA)into the knee joint to establish KOA models.After successful modeling,the rats were continuously treated for five weeks.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of GLP-1,tumor necrosis factor-alpha(TNF-α),and interleukin-1β(IL-1β)in rat serum.Pathological examination was utilized to observe the pathological changes in knee joint cartilage.The mRNA levels of TNF-αand MMP13 in knee joint cartilage were detected by qRT-PCR,and the protein expression levels of GLP-1R,MMP13,and caspase-8 in knee joint cartilage were detected by Western blot.The expression of GLP-1R and MMP13 in the knee joint was detected by immunohistochemistry.Tunel staining was used to observe the apoptosis of chondrocytes in the knee joint.The above experimental results showed that Astragali Radix may raise the serum levels of GLP-1,reduce serum levels of TNF-αand IL-1,and decrease the relative mRNA expression of TNF-αand MMP13 through the GLP-1/GLP-1R axis.It thus activated GLP-1R,reduced the protein expression of MMP13 and caspase-8 in cartilage,and regulated their related signaling pathways to improve inflammation and apoptosis,so as to protect cartilage and improve KOA.

关 键 词：黄芪 膝骨关节炎 软骨保护 GLP-1/GLP-1R轴 

分 类 号：R285.5[医药卫生—中药学]