利拉鲁肽对膝骨关节炎大鼠炎症反应的影响  

Effect of GLP-1 analogue liraglutide on inflammatory response in rat model of knee osteoarthritis

作　　者：史纪元[1] 武世勋[1] 王涛[2] 刘时璋[1] 易智[1] 傅蔷 SHI Jiyuan;WU Shixun;WANG Tao;LIU Shizhang;YI Zhi;FU Qiang(Department of Orthopedics,Shaanxi Provincial People's Hospital,Xi'an,Shaanxi 710068,China;Department of Imaging,Shaanxi Provincial People's Hospital,Xi'an,Shaanxi 710068,China;Department of Hematology and Oncology,The Children's Hospital of Xi'an,Xi'an,Shaanxi 710003,China)

机构地区：[1]陕西省人民医院骨科,陕西西安710068 [2]陕西省人民医院影像科,陕西西安710068 [3]西安市儿童医院血液肿瘤科,陕西西安710003

出　　处：《安徽医药》2022年第2期225-230,F0003,共7页Anhui Medical and Pharmaceutical Journal

基　　金：陕西省重点研发计划项目(2021SF-186、2018ZDXM-SF-057)。

摘　　要：目的探讨胰高血糖素样肽-1(GLP-1)类似物利拉鲁肽对膝骨关节炎大鼠模型炎症反应的改善作用。方法选取成年雄性SD大鼠60只,分为模型组、假手术组和利拉鲁肽组,每组20只。模型组和利拉鲁肽组大鼠通过关节腔内注射单碘乙酸盐(MIA,4 mg)诱导大鼠膝骨关节炎模型。假手术组大鼠关节腔内注射相同体积的生理盐水。拉鲁肽组大鼠皮下注射利拉鲁肽(50μg·kg^(-1)·d^(-1))用于上调GLP-1受体(GLP-1R)的表达,共处理4周。蛋白质印迹法(Westernblotting)检测GLP-1R、蛋白激酶A(PKA)、环状单磷酸腺苷反应元件结合蛋白(CREB)、聚集蛋白聚糖(aggrecan)、Ⅱ型胶原(collagenⅡ)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、基质金属蛋白酶-13(MMP-13)和含Ⅰ型血小板结合蛋白基序的解聚蛋白样金属蛋白酶4(ADAMTS-4)的表达。免疫沉淀技术检测GLP-1R与PKA/CREB信号通路之间的相互作用。结果与假手术组相比,模型组大鼠膝关节软骨组织中GLP-1R的蛋白相对表达量明显下调[(1.12±0.11)比(0.34±0.04)],而利拉鲁肽组(0.87±0.07)明显高于模型组(P<0.05)。与假手术组相比,模型组大鼠膝关节软骨组织中PKA和CREB的磷酸化情况均被显著抑制(P<0.05),利拉鲁肽处理可显著提高PKA和CREB的磷酸化水平(P<0.05)。模型组大鼠膝关节软骨组织中的aggrecan和collagenⅡ蛋白水平显著降低,利拉鲁肽组的两种蛋白的表达均显著升高(P<0.05)。与假手术组相比,模型组大鼠的TNF-α、IL-1β、IL-6、MMP-13和ADAMTS-4蛋白表达水平均显著上调,利拉鲁肽处理可显著抑制膝骨关节炎大鼠膝关节软骨组织中上述因子的上调(P<0.05)。结论利拉鲁肽在膝骨关节炎大鼠模型中具有较好抗炎活性,可显著改善膝关节软骨组织。Objective To investigate the effect of glucagon-like peptide-1(GLP-1)analogue liraglutide on the inflammatory re⁃sponse in rats with knee osteoarthritis.Methods Sixty adult male SD rats were used as the research objects.The rats were divided into a model group,a sham group and a liraglutide group,with 20 rats in each group.The rats in the model group and the liraglutide group were induced by intraperitoneal injection of monoiodoacetate(MIA,4 mg)into the rat knee osteoarthritis model.In the sham group,the same volume of saline was injected into the joint cavity of rats.Rats in the larutide group were injected subcutaneously with liraglutide(50μg·kg^(-1)·d^(-1))to upregulate the expression of GLP-1 receptor(GLP-1R)for 4 weeks.Western blot analysis was used to detect the ex⁃pression of GLP-1R,PKA,CREB,aggrecan,collagenⅡ,TNF-α,IL-1β,IL-6,MMP-13 and ADAMTS-4.Immunoprecipitation was used to detect the interaction between GLP-1R and the PKA/CREB signaling pathway.Results Compared with the sham group,the relative expression of GLP-1R protein in the knee joint cartilage of the model group was significantly down-regulated[(1.12±0.11)vs.(0.34±0.04)],while the liraglutide group(0.87±0.07)was significantly higher than the model group(P<0.05).Compared with the sham group,the phosphorylation of PKA and CREB in the knee joint cartilage of the model group was significantly inhibited(P<0.05),and the treatment with liraglutide significantly increased the phosphorylation level of PKA and CREB(P<0.05).The aggrecan and collagenⅡprotein levels in the knee joint cartilage of the model group were significantly decreased,and the expression of the two proteins in the liraglutide group was significantly increased(P<0.05).Compared with the sham group,the expression levels of TNF-α,IL-1β,IL-6,MMP-13 and ADAMTS-4 protein in the model group were significantly up-regulated,and liraglutide treatment significantly inhibited the up-regulation of the above factors in cartilage tissue(P<0.05).Conclusion Liraglutide has better anti-i

关 键 词：膝骨关节炎 胰高血糖素样肽-1 胰高血糖素样肽-1受体 利拉鲁肽 炎症 软骨 

分 类 号：R684.3[医药卫生—骨科学]