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Metformin alleviates β‑glycerophosphate‑induced calcification of vascular smooth muscle cells via AMPK/mTOR‑activated autophagy

  • Authors:
    • Xiaobo Qiu
    • Qing Xu
    • Tianhua Xu
    • Pengzhi Wan
    • Zitong Sheng
    • Yiran Han
    • Li Yao
  • View Affiliations / Copyright

    Affiliations: Department of Nephrology, The First Hospital of China Medical University, Shenyang, Liaoning 110000, P.R. China, Department of Orthopaedics, Shengjing Hospital of China Medical University, Shenyang, Liaoning 110004, P.R. China
    Copyright: © Qiu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 58
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    Published online on: November 19, 2020
       https://doi.org/10.3892/etm.2020.9490
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Abstract

The aim of the present study was to investigate the effect of metformin on β‑glycerophosphate‑induced calcification of vascular smooth muscle cells (VSMCs) and the possible mechanisms underlying this. Using an established VSMC calcification model, VSMCs were first treated with β‑glycerophosphate, before metformin, 3‑methyladenine and compound C were added to the cell cultures in different combinations. Calcium deposition in the cells was examined by Alizarin Red S staining and using the O‑cresolphthalein complexone method. To assess the occurrence of autophagy, autophagosomes inside the cells were studied using a transmission electron microscope and green fluorescent microtubule‑associated protein 1 light chain 3 (LC3) puncta were examined using a fluorescent microscope. Additionally, protein expression levels of α‑smooth muscle actin (α‑SMA), runt‑related transcription factor 2 (RUNX2), LC3II/I, beclin 1 and 5' adenosine monophosphate‑activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway‑associated proteins were determined by western blot analysis. Metformin increased the number of autophagosomes, green fluorescent LC3 puncta and the levels of LC3II/I, beclin 1, α‑SMA and phosphorylated (p)‑AMPK in the VSMCs that were treated with β‑glycerophosphate when compared to controls; whereas, calcium deposition and the expression levels of RUNX2 and p‑mTOR were found to be decreased. Treating the VSMCs with 3‑methyladenine or compound C reversed the effects of metformin. The results of the present study suggested that metformin may alleviate β‑glycerophosphate‑induced calcification of VSMCs, which may be attributed to the activation of AMPK/mTOR signaling pathway‑dependent autophagy.
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1 

Coresh J: Update on the burden of CKD. J Am Soc Nephrol. 28:1020–1022. 2017.PubMed/NCBI View Article : Google Scholar

2 

Go AS, Chertow GM, Fan D, McCulloch CE and Hsu CY: Chronic kidney disease and the risks of death, cardiovascular events, and hospitalization. N Engl J Med. 351:1296–1305. 2004.PubMed/NCBI View Article : Google Scholar

3 

Bardeesi ASA, Gao J, Zhang K, Yu S, Wei M, Liu P and Huang H: A novel role of cellular interactions in vascular calcification. J Transl Med. 15(95)2017.PubMed/NCBI View Article : Google Scholar

4 

Cannata-Andia JB and Martin KJ: The challenge of controlling phosphorus in chronic kidney disease. Nephrol Dial Transplant. 31:541–547. 2016.PubMed/NCBI View Article : Google Scholar

5 

Durham AL, Speer MY, Scatena M, Giachelli CM and Shanahan CM: Role of smooth muscle cells in vascular calcification: Implications in atherosclerosis and arterial stiffness. Cardiovasc Res. 114:590–600. 2018.PubMed/NCBI View Article : Google Scholar

6 

Tai S, Hu XQ, Peng DQ, Zhou SH and Zheng XL: The roles of autophagy in vascular smooth muscle cells. Int J Cardiol. 211:1–6. 2016.PubMed/NCBI View Article : Google Scholar

7 

Salabei JK and Hill BG: Autophagic regulation of smooth muscle cell biology. Redox Biol. 4:97–103. 2015.PubMed/NCBI View Article : Google Scholar

8 

Nussenzweig SC, Verma S and Finkel T: The role of autophagy in vascular biology. Circ Res. 116:480–488. 2015.PubMed/NCBI View Article : Google Scholar

9 

Dai XY, Zhao MM, Cai Y, Guan QC, Zhao Y, Guan Y, Kong W, Zhu WG, Xu MJ and Wang X: Phosphate-induced autophagy counteracts vascular calcification by reducing matrix vesicle release. Kidney Int. 83:1042–1051. 2013.PubMed/NCBI View Article : Google Scholar

10 

Piwkowska A, Rogacka D, Jankowski M, Dominiczak MH, Stepinski JK and Angielski S: Metformin induces suppression of NAD(P)H oxidase activity in podocytes. Biochem Biophys Res Commun. 393:268–273. 2010.PubMed/NCBI View Article : Google Scholar

11 

Kim J, Kundu M, Viollet B and Guan KL: AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol. 13:132–141. 2011.PubMed/NCBI View Article : Google Scholar

12 

Egan D, Kim J, Shaw RJ and Guan KL: The autophagy initiating kinase ULK1 is regulated via opposing phosphorylation by AMPK and mTOR. Autophagy. 7:643–644. 2011.PubMed/NCBI View Article : Google Scholar

13 

Wang S, Song P and Zou MH: AMP-activated protein kinase, stress responses and cardiovascular diseases. Clin Sci (Lond). 122:555–573. 2012.PubMed/NCBI View Article : Google Scholar

14 

Shioi A, Nishizawa Y, Jono S, Koyama H, Hosoi M and Morii H: Beta-glycerophosphate accelerates calcification in cultured bovine vascular smooth muscle cells. Arterioscler Thromb Vasc Biol. 15:2003–2009. 1995.PubMed/NCBI View Article : Google Scholar

15 

Cao X, Li H, Tao H, Wu N, Yu L, Zhang D, Lu X, Zhu J, Lu Z and Zhu Q: Metformin inhibits vascular calcification in female rat aortic smooth muscle cells via the AMPK-eNOS-NO pathway. Endocrinology. 154:3680–3689. 2013.PubMed/NCBI View Article : Google Scholar

16 

She C, Zhu LQ, Zhen YF, Wang XD and Dong QR: Activation of AMPK protects against hydrogen peroxide-induced osteoblast apoptosis through autophagy induction and NADPH maintenance: New implications for osteonecrosis treatment? Cell Signal. 26:1–8. 2014.PubMed/NCBI View Article : Google Scholar

17 

Zhu D, Mackenzie NC, Shanahan CM, Shroff RC, Farquharson C and MacRae VE: BMP-9 regulates the osteoblastic differentiation and calcification of vascular smooth muscle cells through an ALK1 mediated pathway. J Cell Mol Med. 19:165–174. 2015.PubMed/NCBI View Article : Google Scholar

18 

Shroff R, Long DA and Shanahan C: Mechanistic insights into vascular calcification in CKD. J Am Soc Nephrol. 24:179–189. 2013.PubMed/NCBI View Article : Google Scholar

19 

Bailey CJ: Metformin: Historical overview. Diabetologia. 60:1566–1576. 2017.PubMed/NCBI View Article : Google Scholar

20 

Mary A, Hartemann A, Liabeuf S, Aubert CE, Kemel S, Salem JE, Cluzel P, Lenglet A, Massy ZA, Lalau JD, et al: Association between metformin use and below-the-knee arterial calcification score in type 2 diabetic patients. Cardiovasc Diabetol. 16(24)2017.PubMed/NCBI View Article : Google Scholar

21 

Goldberg RB, Aroda VR, Bluemke DA, Barrett-Connor E, Budoff M, Crandall JP, Dabelea D, Horton ES, Mather KJ, Orchard TJ, et al: Effect of long-term metformin and lifestyle in the diabetes prevention program and its outcome study on coronary artery calcium. Circulation. 136:52–64. 2017.PubMed/NCBI View Article : Google Scholar

22 

Zhang X, Xiao J, Li R, Qin X, Wang F, Mao Y, Liang W, Sheng X, Guo M, Song Y and Ji X: Metformin alleviates vascular calcification induced by vitamin D3 plus nicotine in rats via the AMPK pathway. Vascular pharmacol. 81:83–90. 2016.PubMed/NCBI View Article : Google Scholar

23 

Isoda K, Young JL, Zirlik A, MacFarlane LA, Tsuboi N, Gerdes N, Gerdes N, Schönbeck U and Libby P: Metformin inhibits proinflammatory responses and nuclear factor-kappaB in human vascular wall cells. Arterioscler Thromb Vasc Biol. 26:611–617. 2006.PubMed/NCBI View Article : Google Scholar

24 

Kim SA and Choi HC: Metformin inhibits inflammatory response via AMPK-PTEN pathway in vascular smooth muscle cells. Biochem Biophys Res Commun. 425:866–872. 2012.PubMed/NCBI View Article : Google Scholar

25 

Hao B, Xiao Y, Song F, Long X, Huang J, Tian M, Deng S and Wu Q: Metformin-induced activation of AMPK inhibits the proliferation and migration of human aortic smooth muscle cells through upregulation of p53 and IFI16. Int J Mol Med. 41:1365–1376. 2018.PubMed/NCBI View Article : Google Scholar

26 

Feng Y, He D, Yao Z and Klionsky DJ: The machinery of macroautophagy. Cell Res. 24:24–41. 2014.PubMed/NCBI View Article : Google Scholar

27 

Choi AM, Ryter SW and Levine B: Autophagy in human health and disease. N Engl J Med. 368:651–662. 2013.PubMed/NCBI View Article : Google Scholar

28 

Xie Z, Lau K, Eby B, Lozano P, He C, Pennington B, Li H, Rathi S, Dong Y, Tian R, et al: Improvement of cardiac functions by chronic metformin treatment is associated with enhanced cardiac autophagy in diabetic OVE26 mice. Diabetes. 60:1770–1778. 2011.PubMed/NCBI View Article : Google Scholar

29 

Xu M, Liu L, Song C, Chen W and Gui S: Ghrelin improves vascular autophagy in rats with vascular calcification. Life Sci. 179:23–29. 2017.PubMed/NCBI View Article : Google Scholar

30 

Klionsky DJ, Abdelmohsen K, Abe A, Abedin MJ, Abeliovich H, Acevedo Arozena A, Adachi H, Adams CM, Adams PD, Adeli K, et al: Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy. 12:1–222. 2016.PubMed/NCBI View Article : Google Scholar

31 

Mizushima N, Yoshimori T and Levine B: Methods in mammalian autophagy research. Cell. 140:313–326. 2010.PubMed/NCBI View Article : Google Scholar

32 

Egan DF, Shackelford DB, Mihaylova MM, Gelino S, Kohnz RA, Mair W, Vasquez DS, Joshi A, Gwinn DM, Taylor R, et al: Phosphorylation of ULK1 (hATG1) by AMP-activated protein kinase connects energy sensing to mitophagy. Science. 331:456–461. 2011.PubMed/NCBI View Article : Google Scholar

33 

Webster AC, Nagler EV, Morton RL and Masson P: Chronic kidney disease. Lancet. 389:1238–1252. 2017.PubMed/NCBI View Article : Google Scholar

34 

Neven E, Vervaet B, Brand K, Gottwald-Hostalek U, Opdebeeck B, De Mare A, Verhulst A, Lalau JD, Kamel S, De Broe ME and D'Haese PC: Metformin prevents the development of severe chronic kidney disease and its associated mineral and bone disorder. Kidney Int. 94:102–113. 2018.PubMed/NCBI View Article : Google Scholar

35 

Lipska KJ, Bailey CJ and Inzucchi SE: Use of metformin in the setting of mild-to-moderate renal insufficiency. Diabetes Care. 34:1431–1437. 2011.PubMed/NCBI View Article : Google Scholar

36 

Lalau JD, Kajbaf F, Bennis Y, Hurtel-Lemaire AS, Belpaire F and De Broe ME: Metformin treatment in patients with type 2 diabetes and chronic kidney disease stages 3A, 3B, or 4. Diabetes Care. 41:547–553. 2018.PubMed/NCBI View Article : Google Scholar

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Copy and paste a formatted citation
Spandidos Publications style
Qiu X, Xu Q, Xu T, Wan P, Sheng Z, Han Y and Yao L: Metformin alleviates β‑glycerophosphate‑induced calcification of vascular smooth muscle cells via AMPK/mTOR‑activated autophagy. Exp Ther Med 21: 58, 2021.
APA
Qiu, X., Xu, Q., Xu, T., Wan, P., Sheng, Z., Han, Y., & Yao, L. (2021). Metformin alleviates β‑glycerophosphate‑induced calcification of vascular smooth muscle cells via AMPK/mTOR‑activated autophagy. Experimental and Therapeutic Medicine, 21, 58. https://doi.org/10.3892/etm.2020.9490
MLA
Qiu, X., Xu, Q., Xu, T., Wan, P., Sheng, Z., Han, Y., Yao, L."Metformin alleviates β‑glycerophosphate‑induced calcification of vascular smooth muscle cells via AMPK/mTOR‑activated autophagy". Experimental and Therapeutic Medicine 21.1 (2021): 58.
Chicago
Qiu, X., Xu, Q., Xu, T., Wan, P., Sheng, Z., Han, Y., Yao, L."Metformin alleviates β‑glycerophosphate‑induced calcification of vascular smooth muscle cells via AMPK/mTOR‑activated autophagy". Experimental and Therapeutic Medicine 21, no. 1 (2021): 58. https://doi.org/10.3892/etm.2020.9490
Copy and paste a formatted citation
x
Spandidos Publications style
Qiu X, Xu Q, Xu T, Wan P, Sheng Z, Han Y and Yao L: Metformin alleviates β‑glycerophosphate‑induced calcification of vascular smooth muscle cells via AMPK/mTOR‑activated autophagy. Exp Ther Med 21: 58, 2021.
APA
Qiu, X., Xu, Q., Xu, T., Wan, P., Sheng, Z., Han, Y., & Yao, L. (2021). Metformin alleviates β‑glycerophosphate‑induced calcification of vascular smooth muscle cells via AMPK/mTOR‑activated autophagy. Experimental and Therapeutic Medicine, 21, 58. https://doi.org/10.3892/etm.2020.9490
MLA
Qiu, X., Xu, Q., Xu, T., Wan, P., Sheng, Z., Han, Y., Yao, L."Metformin alleviates β‑glycerophosphate‑induced calcification of vascular smooth muscle cells via AMPK/mTOR‑activated autophagy". Experimental and Therapeutic Medicine 21.1 (2021): 58.
Chicago
Qiu, X., Xu, Q., Xu, T., Wan, P., Sheng, Z., Han, Y., Yao, L."Metformin alleviates β‑glycerophosphate‑induced calcification of vascular smooth muscle cells via AMPK/mTOR‑activated autophagy". Experimental and Therapeutic Medicine 21, no. 1 (2021): 58. https://doi.org/10.3892/etm.2020.9490
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