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

doi:10.13418/j.issn.1001-165x.2017.04.012

中国临床解剖学杂志 ›› 2017, Vol. 35 ›› Issue (4): 413-418.doi: 10.13418/j.issn.1001-165x.2017.04.012

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

谭清梅^1,2，杨诚³ ，廖坚文³ ，邹庆宝⁴，黄爱军⁴，林莉¹ ，张从新¹

1.广州医科大学附属第三医院荔湾医院, 广州 510145; 2.深圳市福田区第二人民医院, 深圳 518049; 3.南方医科大学第三附属医院创伤骨科, 广州510630; 4.中山大学附属第八医院（深圳福田）骨科, 深圳 518033

收稿日期:2017-03-22 出版日期:2017-07-25 发布日期:2017-08-30
通讯作者: 张从新，教授，硕士研究生导师，E-mail: zhang-c-xhappy@163.com
作者简介:共同第一作者：谭清梅（1982-），女，广西桂平人，在读研究生，主要从事全科医学及慢性病的防治研究，Tel:13510457165, E-mail: 87810465@qq.com；杨诚（1981-），男，广东湛江人，博士，主要从事创伤修复及骨退行性病变，E-mail: 292990433@qq.com
基金资助:
深圳市福田区卫生公益性科研项目（Ftws2017011)

Metformin protects osteoarthritis in mice by regulating the AMPK signaling pathways

TAN Qing-mei ^1,2, YANG Cheng³, LIAO Jian-wen³, Zou Qingbao⁴， Huang Aijun⁴， LIN Li¹, ZHANG Cong-xin¹

1.The Liwan Hospital of the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510145, China; 2.The Second People's Hospital of Futian District Shenzhen, Shenzhen 518049, China; 3.Department of Traumatic Surgery, The Third affiliated Hospital of Southern Medical University ,Guangzhou, 510630,China; 4.Department of Orthopedics, The Eighth affiliated Hospital of Sun Yat-Sen University ,Shenzhen 518033, China

Received:2017-03-22 Online:2017-07-25 Published:2017-08-30

摘要/Abstract

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、Micro-CT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。

关键词: 二甲双胍, , , 骨关节炎, , AMPK, , , mTOR, , 自噬

Abstract:

Objective　Study the effect of metformin on osteoarthritis and chondrocyte differentiation in mice.　Methods　Mice were divided into a control group and an experimental group whose anterior cruciate ligament and medial meniscus excision were done in the right knee of both groups for arthritis model; Aftersuccess of model establishment, the experimental group was given metformin (2mg/kg/d) and the control group was given saline as intervention treatment for 8 weeks; Then through the HE, sarranine O-fast green slice staining and Micro-CT structural change was observed and grading was performed according to Mankin ratings; The AMPK, mTOR activity and degree of autophagy of articular cartilage in two groups were tested by Western Blot; Chondrocytes were cultivated in vitro and stimulated by metformin to observe the cell morphology, and amount. The cell activity was determined by MTT method .The cell activity of AMPK, mTOR and autophagy were detected by Western Blot.　Results　(1)We succeeded in establishing the model of osteoarthritis in mice and found metformin could protect the osteoarthritis by Micro - CT, HE and sarranine O-fast green slice staining and Mankin ratings. (2) Western Blot results showed metformin activated AMPK and enhanced autophagy to protect osteoarthritis. (3) MTT method showed metformin inhibited Chondrocytes differentiation,and Western Blot results showed metformin promoted chondrocytes autophagy.　Conclusions　Metformin activates AMPK signaling pathways, inhibits mTOR signaling pathways, protecting chondrocyte autophagy，thus protects osteoarthritis in mice.

Key words: Metformin; , Osteoarthritis; , AMPK; , mTOR; , Autophagy

谭清梅，杨诚，廖坚文，邹庆宝，黄爱军，林莉，张从新. 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[J]. 中国临床解剖学杂志, 2017, 35(4): 413-418.

TAN Qing-mei, YANG Cheng, LIAO Jian-wen, Zou Qingbao， Huang Aijun， LIN Li, ZHANG Cong-xin. Metformin protects osteoarthritis in mice by regulating the AMPK signaling pathways[J]. Chinese Journal Of Clinical Anatomy, 2017, 35(4): 413-418.

参考文献

[1] Takayama K, Ishida K, Matsushita T, et al. SIRT1 regulation of apoptosis of human chondrocytes[J]. Arthritis Rheum, 2009, 60(9):2731-2740.
[2] Hardie D G. AMP-activated/SNF1 protein kinases: conserved guardians of cellular energy[J]. Nat Rev Mol Cell Biol, 2007, 8(10):774-785.
[3] Shah M, Kola B, Bataveljic A, et al. AMP-activated protein kinase (AMPK) activation regulates in vitro bone formation and bone mass[J]. Bone, 2010, 47(2):309-319.
[4] Mai Q G, Zhang Z M, Xu S, et al. Metformin stimulates osteoprotegerin and reduces RANKL expression in osteoblasts and ovariectomized rats[J]. J Cell Biochem, 2011, 112(10):2902-2909.
[5] Jeyabalan J, Shah M, Viollet B, et al. Mice lacking AMP-activated protein kinase alpha1 catalytic subunit have increased bone remodelling and modified skeletal responses to hormonal challenges induced by ovariectomy and intermittent PTH treatment[J]. J Endocrinol, 2012, 214(3):349-358.
[6] Quinn J M, Tam S, Sims N A, et al. Germline deletion of AMP-activated protein kinase beta subunits reduces bone mass without altering osteoclast differentiation or function[J]. FASEB J, 2010, 24(1):275-285.
[7] Terkeltaub R, Yang B, Lotz M, et al. Chondrocyte AMP-activated protein kinase activity suppresses matrix degradation responses to proinflammatory cytokines interleukin-1beta and tumor necrosis factor alpha[J]. Arthritis Rheum, 2011, 63(7):1928-1937.
[8] Zhang Y, Vasheghani F, Li Y H, et al. Cartilage-specific deletion of mTOR upregulates autophagy and protects mice from osteoarthritis[J]. Ann Rheum Dis, 2015, 74(7):1432-1440.
[9] Yang C, Li Z, Lai P, et al. Chondrocyte-Specific Ablation of AMPKalpha1 Does Not Affect Bone Development or Pathogenesis of Osteoarthritis in Mice[J]. DNA Cell Biol, 2016, 35(3):156-162.
[10] Zhou S, Lu W, Chen L, et al. AMPK deficiency in chondrocytes accelerated the progression of instability-induced and ageing-associated osteoarthritis in adult mice[J]. Sci Rep, 2017, 7:43245.
[11] Gao Y, Li Y, Xue J, et al. Effect of the anti-diabetic drug metformin on bone mass in ovariectomized rats[J]. Eur J Pharmacol, 2010, 635(1-3):231-236.
[12] Mankin H J. Biochemical and metabolic aspects of osteoarthritis[J]. Orthop Clin North Am, 1971, 2(1):19-31.
[13] Matsuzaki T, Matsushita T, Tabata Y, et al. Intra-articular administration of gelatin hydrogels incorporating rapamycin-micelles reduces the development of experimental osteoarthritis in a murine model[J]. Biomaterials, 2014, 35(37):9904-9911.
[14] Takayama K, Kawakami Y, Kobayashi M, et al. Local intra-articular injection of rapamycin delays articular cartilage degeneration in a murine model of osteoarthritis[J]. Arthritis Res Ther, 2014, 16(6):482.
[15] Cravedi P, Ruggenenti P, Remuzzi G. Sirolimus to replace calcineurin inhibitors? Too early yet[J]. Lancet, 2009, 373(9671):1235-1236.
[16] Bandow K, Kusuyama J, Kakimoto K, et al. AMP-activated protein kinase (AMPK) activity negatively regulates chondrogenic differentiation[J]. Bone, 2015, 74:125-133.
[17] Mccarty M F. AMPK activation--protean potential for boosting healthspan[J]. Age (Dordr), 2014, 36(2):641-663.

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

Metformin protects osteoarthritis in mice by regulating the AMPK signaling pathways

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