Skip to main content

Advertisement

SpringerLink
Log in

The protective effects of metformin in an in vitro model of aging 3T3 fibroblast under the high glucose conditions

  • Original Article
  • Published:
Journal of Physiology and Biochemistry Aims and scope Submit manuscript

Abstract

Metformin is the most widely used anti-diabetic drug in the world. It reduces advanced glycation end product (AGEs)-induced ROS generation in high glucose condition. Protein glycation contributes to skin aging as it deteriorates the existing collagen by crosslinking. The progressive increase of AGE during aging not only causes oxidative damage to cellular macromolecules but also modulates the activation of transcription factors nuclear factor kappa-B(NF-kB). However, it is still unclear whether metformin can change collagen production and NF-kB activity induced by high glucose conditions in 3T3 fibroblast. The effects of metformin on proliferation, apoptosis, and collagen levels and NF-kB activity of in vitro cell aging model of 3T3 fibroblast cells in high glucose conditions. At first, we investigated the effects of 50 mM high glucose concentration, with or without metformin, on 3T3 fibroblast proliferation, by BrdU immunostaining for cell proliferation. Apoptotic levels were analyzed by flow cytometric assay. NF-kB(p65) activity was measured by transcription factor assay kit and collagen I and III levels by Collagen Estimation Assay through ELISA. We observed that metformin exposure leads to decreased apoptosis levels and increased proliferation of 3T3 fibroblast in high glucose media. We also determined that metformin exposure leads to increased production of collagen I–III and decreased activation of NF-kB(p65) activity. The data are consistent with the observation that metformin has a protective effect in this in vitro model of aging 3T3 fibroblasts under high glucose conditions inducing cell proliferation, collagen I and III production, protection from apoptosis, and reducing NF-kB(p65) activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Beauchef G, Bigot N, Kypriotou M, Renard E, Porée B, Widom R, Dompmartin-Blanchere A, Oddos T, Maquart FX, Demoor M, Boumediene K, Galera P (2012) The p65 subunit of NF-κB inhibits COL1A1 gene transcription in human dermal and scleroderma fibroblasts through its recruitment on promoter by protein interaction with transcriptional activators (c-Krox, Sp1, and Sp3). J Biol Chem 287:3462–3478

    Article  CAS  Google Scholar 

  2. Bellin C, de Wiza DH, Wiernsperger NF et al (2006) Generation of reactive oxygen species by endothelial and smooth muscle cells: influence of hyperglycemia and metformin. Horm Metab Res 38:732–739

    Article  CAS  Google Scholar 

  3. Benazzoug Y, Borchiellini C, Labat-Robert J, Robert L, Kern P (1998) Effect of high-glucose concentrations on the expression of collagens and fibronectin by fibroblasts in culture. Exp Gerontol 33:445–455

    Article  CAS  Google Scholar 

  4. Bigot N, Beauchef G, Hervieu M, Oddos T, Demoor M, Boumediene K, Galéra P (2012) NF-κB accumulation associated with COL1A1 transactivators defects during chronological aging represses type I collagen expression through a −112/−61-bp region of the COL1A1 promoter in human skin fibroblasts. J Invest Dermatol 132:2360–2367

    Article  CAS  Google Scholar 

  5. Bond M, Baker AH, Newby AC (1999) Nuclear factor kappaB activity is essential for matrix metalloproteinase-1 and -3 upregulation in rabbit dermal fibroblasts. Biochem Biophys Res Commun 264:561–567

    Article  CAS  Google Scholar 

  6. Bucala R, Cerami A (1992) Advanced glycosylation: chemistry, biology, and implications for diabetes and aging. Adv Pharmacol 23:1–34

    Article  CAS  Google Scholar 

  7. Campbell KJ, Perkins ND (2006) Regulation of NF-kappaB function. Biochem Soc Symp:165–180

  8. Dasu MR, Barrow RE, Spies M, Herndon DN (2003) Matrix metalloproteinase expression in cytokine stimulated human dermal fibroblasts. Burns 29:527–531

    Article  Google Scholar 

  9. Donghu Z, Yirong C, Xulei T (2010) Metformin reverses the deleterious effects of high glucose on osteoblast function. J Diabetes Complicat 24:334–344

    Article  Google Scholar 

  10. Dyer DG, Dunn JA, Thorpe SR et al (1993) Accumulation of Maillard reaction products in skin collagen in diabetes and aging. J Clin Invest 91:2463–2469

    Article  CAS  Google Scholar 

  11. Gallo A, Ceolotto G, Pinton P, Iori E, Murphy E, Rutter GA, Rizzuto R, Semplicini A, Avogaro A (2005) Metformin prevents glucose-induced protein kinase C-beta2 activation in human umbilical vein endothelial cells through an antioxidant mechanism. Diabetes 54:1123–1131

    Article  CAS  Google Scholar 

  12. Gkogkolou P, Böhm M (2012) Advanced glycation end products: key players in skin aging? Dermatoendocrinology 4:259–270

    Article  CAS  Google Scholar 

  13. Hong MJ, Ko EB, Park SK, Chang MS (2013) Inhibitory effect of Astragalus membranaceus root on matrix metalloproteinase-1 collagenase expression and procollagen destruction in ultraviolet B-irradiated human dermal fibroblasts by suppressing nuclear factor kappa-B activity. J Pharm Pharmacol 65:142–148

    Article  CAS  Google Scholar 

  14. Jimenez F, Mitts TF, Liu K (2006) Ellagic and tannic acids protect newly synthesized elastic fibers from premature enzymatic degradation in dermal fibroblast cultures. J Invest Dermatol 126:1272–1280

    Article  CAS  Google Scholar 

  15. Kanigur-Sultuybek G, Güven M, Onaran I (1995) The effect of metformin on insulin receptors and lipid peroxidation in alloxan and streptozotocin induced diabetes. J Basic Clin Physiol Pharmacol 6:271–280

    Article  CAS  Google Scholar 

  16. Kanigür-Sultuybek G, Ozdas SB, Curgunlu A, Tezcan V, Onaran I (2007) Does metformin prevent shortterm oxidant-induced dna damage? In vitro study on lymphocytes from aged subjects. J Basic Clin Physiol Pharmacol 18:129–140

    Article  Google Scholar 

  17. Kido D, Mizutani K, Takeda K, Mikami R, Matsuura T, Iwasaki K, Izumi Y (2017) Impact of diabetes on gingival wound healing via oxidative stress. PLoS One 21:1–22

    Google Scholar 

  18. Lu P, Takai K, Weaver VM, Werb Z (2011) Extracellular matrix degradation and remodeling in development and disease. Cold Spring Harb Perspect Biol 1:3(12)

    Google Scholar 

  19. Monnier VM, Glomb M, Elgawish A, Sell DR (1996) The mechanism of collagen cross-linking in diabetes: a puzzle nearing resolution. Diabetes 45:67–72

    Article  Google Scholar 

  20. Oikarinen A (2004) Connective tissue and ageing. Int J Cosmet Sci 26:107–108

    Article  Google Scholar 

  21. Onaran I, Guven GS, Ozdaş SB, Kanigur G, Vehid S (2006) Metformin does not prevent DNA damage in lymphocytes despite its antioxidant properties against cumene hydroperoxide-induced oxidative stress. Mutat Res 611:1–8

    Article  CAS  Google Scholar 

  22. Rinnerthaler M, Bischof J, Karolin-Streubel M (2015) Oxidative stress in aging human skin. Biomol Ther 5:545–589

    CAS  Google Scholar 

  23. Rippe RA, Schrum LW, Stefanovic B, Solís-Herruzo JA, Brenner DA (1999) NF-kappaB inhibits expression of the alpha1(I) collagen gene. DNA Cell Biol 18:751–761

    Article  CAS  Google Scholar 

  24. Schurman L, McCarthy AD, Sedlinsky C, Gangoiti MV, Arnol V, Bruzzone L, Cortizo AM (2008) Metformin reverts deleterious effects of advanced glycation end-products (AGEs) on osteoblastic cells. Exp Clin Endocrinol Diabetes 116:333–340

    Article  CAS  Google Scholar 

  25. Shao X, Cao X, Song G, Zhao Y, Shi B (2014) Metformin rescues the MG63 osteoblasts against the effect of high glucose on proliferation. J Diabetes Res 2014:453940

    Article  Google Scholar 

  26. Singh R, Barden A, Mori T, Beilin L (2001) Advanced glycation end-products: a review. Diabetologia 44:129–146

    Article  CAS  Google Scholar 

  27. Sun LQ, Zhao J, Zhang TT (2012) Protective effects of Salvianolic acid B on Schwann cells apoptosis induced by high glucose. Neurochem Res 37:996–1010

    Article  CAS  Google Scholar 

  28. Trevisan R, Yip J, Sarika L, Li LK, Viberti G (1997) Enhanced collagen synthesis in cultured skin fibroblasts from insulin-dependent diabetic patients with nephropathy. J Am Soc Nephrol 8:1133–1139

    CAS  PubMed  Google Scholar 

  29. Uslu B, Biltekin B, Denir S (2015) Differences between solution and membrane forms of chitosan on the in vitro activity of fibroblasts Balkan. Med J 32:69–78

    CAS  Google Scholar 

  30. Varani J, Dame MK, Rittie L, Fligiel SE, Kang S, Fisher GJ, Voorhees JJ (2006) Decreased collagen production in chronologically aged skin: roles of age-dependent alteration in fibroblast function and defective mechanical stimulation. Am J Pathol 168:1861–1868

    Article  CAS  Google Scholar 

  31. Visse R, Nagase H (2003) Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function, and biochemistry. Circ Res 92:827–839

    Article  CAS  Google Scholar 

  32. Wiernsperger NF (2000) Metformin: intrinsic vasculoprotective properties. Diabetes Technol Ther 2:259–272

    Article  CAS  Google Scholar 

  33. Willershausen-Zönnchen B, Lemmen C, Hamm G (1991) Influence of high glucose concentrations on glycosaminoglycan and collagen synthesis in cultured human gingival fibroblasts. J Clin Periodontol 18:190–195

    Article  Google Scholar 

Download references

Acknowledgements

This work was supported by Scientific Research Project Coordination Unit of Istanbul University, Turkey [Project Number 21204]. The authors thank MSc. Burcu Biltekin, PhD student of cell culture laboratory in Istanbul University, Cerrahpasa Medical Faculty, Histology and Embryology Department.

Author information

Authors and Affiliations

  1. Department of Medical Biology, Cerrahpasa Faculty of Medicine, Istanbul University, Istanbul, Turkey

    Tugba Soydas & Matem Tuncdemir

  2. Department of Histology and Embryology, Istanbul University, Istanbul Faculty of Medicine, Istanbul, Turkey

    Elif Yaprak Sarac, Sibel Dogan & Seyhun Solakoglu

  3. Department of Immunology, Istanbul University, Institute of Experimental Medicine, Istanbul, Turkey

    Suzan Cinar

  4. Department of Medical Biology and Genetics, Istanbul Aydin University, Medical Faculty, 34295 Sefakoy-Kucukcekmece, Istanbul, Turkey

    Gonul Kanigur Sultuybek

Authors
  1. Tugba Soydas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Elif Yaprak Sarac
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Suzan Cinar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Sibel Dogan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Seyhun Solakoglu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Matem Tuncdemir
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Gonul Kanigur Sultuybek
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gonul Kanigur Sultuybek.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Human and animal rights

The article does not contain any studies with human participants or animals performed by any of the authors.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Soydas, T., Yaprak Sarac, E., Cinar, S. et al. The protective effects of metformin in an in vitro model of aging 3T3 fibroblast under the high glucose conditions. J Physiol Biochem 74, 273–281 (2018). https://doi.org/10.1007/s13105-018-0613-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13105-018-0613-5

Keywords

Search

Navigation