二甲双胍通过AMPK信号通路对小鼠骨关节炎保护作用的研究
发布时间：2018-08-10 22:11
【摘要】：目的研究二甲双胍对小鼠骨关节炎及软骨细胞分化的影响。方法切除小鼠右膝关节的内侧半月板和前交叉韧带造模骨关节炎,造模成功后,随机分为2组,实验组给予二甲双胍(2mg/kg/d)而对照组予等量的生理盐水灌胃干预治疗8周,通过HE、番红O-快速绿切片染色和Micro-CT观察关节结构变化,进行Mankin评分评估关节炎严重程度,通过Western Blot检测各组关节软骨AMPK、mTOR的活性及自噬程度;取新生小鼠肋软骨和膝关节软骨进行细胞培养,随机分为实验组(2mmol/L的二甲双胍)和空白对照组,观察软骨细胞形态、数量,MTT法检测细胞活性,Western Blot检测软骨细胞AMPK、mTOR的活性及自噬程度。结果 (1)成功构建了小鼠骨关节炎模型,HE、番红O-快速绿染色、MicroCT结果及Mankin评分证明二甲双胍能保护骨关节炎;(2)关节软骨Western Blot结果显示二甲双胍能激活AMPK,增强自噬保护骨关节炎;(3)MTT法显示二甲双胍抑制软骨细胞增殖分化,Western Blot结果显示二甲双胍促进软骨细胞自噬。结论二甲双胍通过激活AMPK信号通路,抑制mTOR信号通路,促进软骨细胞自噬,发挥保护骨关节炎作用。
[Abstract]:Objective to study the effect of metformin on osteoarthritis and chondrocyte differentiation in mice. Methods the medial meniscus and anterior cruciate ligament (ACL) were excised from the right knee joint of mice. After successful modeling, the mice were randomly divided into two groups. The experimental group was treated with metformin (2mg/kg/d) and the control group was treated with the same amount of normal saline for 8 weeks. The changes of articular structure were observed by Micro-CT, and the severity of arthritis was evaluated by Mankin score. The activity of AMPK mTOR and the degree of autophagy of articular cartilage were detected by Western Blot. The chondrocytes were cultured in the costal cartilage and knee cartilage of newborn mice and were randomly divided into two groups: the experimental group (metformin of 2mmol/L) and the blank control group. The morphology of chondrocytes was observed. The activity of AMPK mTOR and the degree of autophagy in chondrocytes were detected by quantitative MTT assay and Western Blot. Results (1) A mouse model of osteoarthritis was successfully constructed. The results of MicroCT and Mankin score showed that metformin could protect osteoarthritis, and (2) Western Blot of articular cartilage showed that metformin could activate AMPK and enhance autophagy. (3) MTT assay showed that metformin inhibited the proliferation and differentiation of chondrocytes. Western Blot showed that metformin promoted autophagy of chondrocytes. Conclusion Metformin activates AMPK signaling pathway, inhibits mTOR signaling pathway, promotes chondrocyte autophagy and protects osteoarthritis.
【作者单位】： 广州医科大学附属第三医院荔湾医院;深圳市福田区第二人民医院;南方医科大学第三附属医院创伤骨科;中山大学附属第八医院(深圳福田)骨科;
【基金】：深圳市福田区卫生公益性科研项目(Ftws2017011)
【分类号】：R684.3
本文编号：2176357
[Abstract]:Objective to study the effect of metformin on osteoarthritis and chondrocyte differentiation in mice. Methods the medial meniscus and anterior cruciate ligament (ACL) were excised from the right knee joint of mice. After successful modeling, the mice were randomly divided into two groups. The experimental group was treated with metformin (2mg/kg/d) and the control group was treated with the same amount of normal saline for 8 weeks. The changes of articular structure were observed by Micro-CT, and the severity of arthritis was evaluated by Mankin score. The activity of AMPK mTOR and the degree of autophagy of articular cartilage were detected by Western Blot. The chondrocytes were cultured in the costal cartilage and knee cartilage of newborn mice and were randomly divided into two groups: the experimental group (metformin of 2mmol/L) and the blank control group. The morphology of chondrocytes was observed. The activity of AMPK mTOR and the degree of autophagy in chondrocytes were detected by quantitative MTT assay and Western Blot. Results (1) A mouse model of osteoarthritis was successfully constructed. The results of MicroCT and Mankin score showed that metformin could protect osteoarthritis, and (2) Western Blot of articular cartilage showed that metformin could activate AMPK and enhance autophagy. (3) MTT assay showed that metformin inhibited the proliferation and differentiation of chondrocytes. Western Blot showed that metformin promoted autophagy of chondrocytes. Conclusion Metformin activates AMPK signaling pathway, inhibits mTOR signaling pathway, promotes chondrocyte autophagy and protects osteoarthritis.
【作者单位】： 广州医科大学附属第三医院荔湾医院;深圳市福田区第二人民医院;南方医科大学第三附属医院创伤骨科;中山大学附属第八医院(深圳福田)骨科;
【基金】：深圳市福田区卫生公益性科研项目(Ftws2017011)
【分类号】：R684.3
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