Skip to main content
SpringerLink
Log in

Overexpression of CTRP9 attenuates the development of atherosclerosis in apolipoprotein E-deficient mice

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

This study was aimed to explore the role of C1q/TNF-related protein 9 (CTRP9) on atherosclerotic lesion formation. A recombinant lentiviral vector carrying mouse CTRP9 (Lv-CTRP9) was injected intravenously into apolipoprotein E knockout (ApoE−/−) mice given a high-fat diet (HFD). CTRP9 overexpression substantially attenuated atherosclerotic lesion size of mice. The accumulation of macrophages and smooth muscle cells (SMCs) was significantly decreased in atherosclerotic regions with CTRP9 overexpression by immunohistochemical analysis. In addition, CTRP9 downregulated the expressions of monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor-alpha (TNF-α), two main proinflammatory cytokines in atherosclerosis. Furthermore, the autophagy level remarkably increased which was presented by microtubule-associated protein light chain 3B (LC3B) conversion and sequestosome 1 (SQSTM1/p62) degradation. Further study showed that CTRP9 increased adenosine monophosphate-activated protein kinase (AMPK) phosphorylation and decreased mammalian target of rapamycin (mTOR) phosphorylation in vivo. These observations reveal that CTRP9 exerts a protecting role in early atherosclerotic lesions and its anti-atherosclerotic effect is associated with autophagy induction through AMPK/mTOR signaling pathway.

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

Similar content being viewed by others

References

  1. Pagidipati NJ, Gaziano TA (2013) Estimating deaths from cardiovascular disease: a review of global methodologies of mortality measurement. Circulation 127:749–756

    Article  PubMed  PubMed Central  Google Scholar 

  2. Libby P (2002) Inflammation in atherosclerosis. Nature 420:868–874

    Article  CAS  PubMed  Google Scholar 

  3. Stoll G, Bendszus M (2006) Inflammation and atherosclerosis: novel insights into plaque formation and destabilization. Stroke 37:1923–1932

    Article  CAS  PubMed  Google Scholar 

  4. Moore KJ, Tabas I (2011) Macrophages in the pathogenesis of atherosclerosis. Cell 145:341–355

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Mudau M, Genis A, Lochner A, Strijdom H (2012) Endothelial dysfunction: the early predictor of atherosclerosis. Cardiovasc J Afr 23:222–231

    Article  PubMed  PubMed Central  Google Scholar 

  6. Bennett MR, Sinha S, Owens GK (2016) Vascular smooth muscle cells in atherosclerosis. Circ Res 118:692–702

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Jawien J, Nastalek P, Korbut R (2004) Mouse models of experimental atherosclerosis. J Physiol Pharmacol 55:503–517

    CAS  PubMed  Google Scholar 

  8. Meir KS, Leitersdorf E (2004) Atherosclerosis in the apolipoprotein-E-deficient mouse: a decade of progress. Arterioscler Thromb Vasc Biol 24:1006–1014

    Article  CAS  PubMed  Google Scholar 

  9. Nakashima Y, Plump AS, Raines EW, Breslow JL, Ross R (1994) ApoE-deficient mice develop lesions of all phases of atherosclerosis throughout the arterial tree. Arterioscler Thromb 14:133–140

    Article  CAS  PubMed  Google Scholar 

  10. Mizushima N, Komatsu M (2011) Autophagy: renovation of cells and tissues. Cell 147:728–741

    Article  CAS  PubMed  Google Scholar 

  11. Schrijvers DM, De Meyer GR, Martinet W (2011) Autophagy in atherosclerosis: a potential drug target for plaque stabilization. Arterioscler Thromb Vasc Biol 31:2787–2791

    Article  CAS  PubMed  Google Scholar 

  12. Mei Y, Thompson MD, Cohen RA, Tong X (2015) Autophagy and oxidative stress in cardiovascular diseases. Biochim Biophys Acta 1852:243–251

    Article  CAS  PubMed  Google Scholar 

  13. Muller C, Salvayre R, Negre-Salvayre A, Vindis C (2011) Oxidized LDLs trigger endoplasmic reticulum stress and autophagy: prevention by HDLs. Autophagy 7:541–543

    Article  CAS  PubMed  Google Scholar 

  14. De Meyer GR, Grootaert MO, Michiels CF, Kurdi A, Schrijvers DM, Martinet W (2015) Autophagy in vascular disease. Circ Res 116:468–479

    Article  CAS  PubMed  Google Scholar 

  15. Liao X, Sluimer JC, Wang Y, Subramanian M, Brown K, Pattison JS, Robbins J, Martinez J, Tabas I (2012) Macrophage autophagy plays a protective role in advanced atherosclerosis. Cell Metab 15:545–553

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Razani B, Feng C, Coleman T, Emanuel R, Wen H, Hwang S, Ting JP, Virgin HW, Kastan MB, Semenkovich CF (2012) Autophagy links inflammasomes to atherosclerotic progression. Cell Metab 15:534–544

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Le Guezennec X, Brichkina A, Huang YF, Kostromina E, Han W, Bulavin DV (2012) Wip1-dependent regulation of autophagy, obesity, and atherosclerosis. Cell Metab 16:68–80

    Article  CAS  PubMed  Google Scholar 

  18. Sengupta S, Peterson TR, Sabatini DM (2010) Regulation of the mTOR complex 1 pathway by nutrients, growth factors, and stress. Mol Cell 40:310–322

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Pakala R, Stabile E, Jang GJ, Clavijo L, Waksman R (2005) Rapamycin attenuates atherosclerotic plaque progression in apolipoprotein E knockout mice: inhibitory effect on monocyte chemotaxis. J Cardiovasc Pharmacol 46:481–486

    Article  CAS  PubMed  Google Scholar 

  20. Baetta R, Granata A, Canavesi M, Ferri N, Arnaboldi L, Bellosta S, Pfister P, Corsini A (2009) Everolimus inhibits monocyte/macrophage migration in vitro and their accumulation in carotid lesions of cholesterol-fed rabbits. J Pharmacol Exp Ther 328:419–425

    Article  CAS  PubMed  Google Scholar 

  21. Zhao L, Ding T, Cyrus T, Cheng Y, Tian H, Ma M, Falotico R, Pratico D (2009) Low-dose oral sirolimus reduces atherogenesis, vascular inflammation and modulates plaque composition in mice lacking the LDL receptor. Br J Pharmacol 156:774–785

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Wang X, Li L, Li M, Dang X, Wan L, Wang N, Bi X, Gu C, Qiu S, Niu X, Zhu X, Wang L (2013) Knockdown of mTOR by lentivirusmediated RNA interference suppresses atherosclerosis and stabilizes plaques via a decrease of macrophages by autophagy in apolipoprotein Edeficient mice. Int J Mol Med 32:1215–1221

    Article  CAS  PubMed  Google Scholar 

  23. Inoki K, Kim J, Guan KL (2012) AMPK and mTOR in cellular energy homeostasis and drug targets. Annu Rev Pharmacol Toxicol 52:381–400

    Article  CAS  PubMed  Google Scholar 

  24. Wong GW, Krawczyk SA, Kitidis-Mitrokostas C, Ge G, Spooner E, Hug C, Gimeno R, Lodish HF (2009) Identification and characterization of CTRP9, a novel secreted glycoprotein, from adipose tissue that reduces serum glucose in mice and forms heterotrimers with adiponectin. Faseb j 23:241–258

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Peterson JM, Wei Z, Seldin MM, Byerly MS, Aja S, Wong GW (2013) CTRP9 transgenic mice are protected from diet-induced obesity and metabolic dysfunction. Am J Physiol Regul Integr Comp Physiol 305:R522–R533

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Wei Z, Lei X, Petersen PS, Aja S, Wong GW (2014) Targeted deletion of C1q/TNF-related protein 9 increases food intake, decreases insulin sensitivity, and promotes hepatic steatosis in mice. Am J Physiol Endocrinol Metab 306:E779–E790

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Shibata R, Ouchi N, Ohashi K, Murohara T (2017) The role of adipokines in cardiovascular disease. J Cardiol 70:329–334

    Article  PubMed  Google Scholar 

  28. Wang J, Hang T, Cheng XM, Li DM, Zhang QG, Wang LJ, Peng YP, Gong JB (2015) Associations of C1q/TNF-related protein-9 levels in serum and epicardial adipose tissue with coronary atherosclerosis in humans. Biomed Res Int 2015:971683

  29. Li J, Zhang P, Li T, Liu Y, Zhu Q, Chen T, Liu T, Huang C, Zhang J, Zhang Y, Guo Y (2015) CTRP9 enhances carotid plaque stability by reducing pro-inflammatory cytokines in macrophages. Biochem Biophys Res Commun 458:890–895

    Article  CAS  PubMed  Google Scholar 

  30. Jung TW, Hong HC, Hwang HJ, Yoo HJ, Baik SH, Choi KM (2015) C1q/TNF-Related Protein 9 (CTRP9) attenuates hepatic steatosis via the autophagy-mediated inhibition of endoplasmic reticulum stress. Mol Cell Endocrinol 417:131–140

    Article  CAS  PubMed  Google Scholar 

  31. Lin Y, Bai L, Chen Y, Zhu N, Bai Y, Li Q, Zhao S, Fan J, Liu E (2015) Practical assessment of the quantification of atherosclerotic lesions in apoE(-)/(-) mice. Mol Med Rep 12:5298–5306

    Article  CAS  PubMed  Google Scholar 

  32. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  33. Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc 3:1101–1108

    Article  CAS  PubMed  Google Scholar 

  34. Uemura Y, Shibata R, Ohashi K, Enomoto T, Kambara T, Yamamoto T, Ogura Y, Yuasa D, Joki Y, Matsuo K, Miyabe M, Kataoka Y, Murohara T, Ouchi N (2013) Adipose-derived factor CTRP9 attenuates vascular smooth muscle cell proliferation and neointimal formation. Faseb J 27:25–33

    Article  CAS  PubMed  Google Scholar 

  35. Kambara T, Shibata R, Ohashi K, Matsuo K, Hiramatsu-Ito M, Enomoto T, Yuasa D, Ito M, Hayakawa S, Ogawa H, Aprahamian T, Walsh K, Murohara T, Ouchi N (2015) C1q/Tumor necrosis factor-related protein 9 protects against acute myocardial injury through an adiponectin receptor I-AMPK-dependent mechanism. Mol Cell Biol 35:2173–2185

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Legein B, Temmerman L, Biessen EA, Lutgens E (2013) Inflammation and immune system interactions in atherosclerosis. Cell Mol Life Sci 70:3847–3869

    Article  CAS  PubMed  Google Scholar 

  37. Fatkhullina AR, Peshkova IO, Koltsova EK (2016) The role of cytokines in the development of atherosclerosis. Biochemistry 81:1358–1370

    CAS  PubMed  Google Scholar 

  38. Li Y, Geng X, Wang H, Cheng G, Xu S (2016) CTRP9 ameliorates pulmonary-arterial hypertension through attenuating inflammation and improving endothelial cell survival and function. J Cardiovasc Pharmacol 67:394–401

    Article  CAS  PubMed  Google Scholar 

  39. Jung CH, Lee MJ, Kang YM, Lee Y, Seol SM, Yoon HK, Kang SW, Lee WJ, Park JY (2015) C1q/TNF-related protein-9 inhibits cytokine-induced vascular inflammation and leukocyte adhesiveness via AMP-activated protein kinase activation in endothelial cells. Mol Cell Endocrinol 419:235–243

    Article  CAS  PubMed  Google Scholar 

  40. Zhang P, Huang C, Li J, Li T, Guo H, Liu T, Li N, Zhu Q, Guo Y (2016) Globular CTRP9 inhibits oxLDL-induced inflammatory response in RAW 264.7 macrophages via AMPK activation. Mol Cell Biochem 417:67–74

    Article  CAS  PubMed  Google Scholar 

  41. Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T (2000) LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. Embo J 19:5720–5728

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Evans TD, Sergin I, Zhang X, Razani B (2017) Target acquired: Selective autophagy in cardiometabolic disease. Sci Signal 10:eaag2298

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. He C, Klionsky DJ (2009) Regulation mechanisms and signaling pathways of autophagy. Annu Rev Genet 43:67–93

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Levine B, Mizushima N, Virgin HW (2011) Autophagy in immunity and inflammation. Nature 469:323–335

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Lapaquette P, Guzzo J, Bretillon L, Bringer MA (2015) Cellular and molecular connections between autophagy and inflammation. Mediat Inflamm 2015:398483

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant No. 81671950) and Department of Science and Technology of Shandong Province (2014GSF118020).

Author information

Authors and Affiliations

  1. Department of Cardiology, Qilu Hospital of Shandong University, Jinan, 250012, Shandong, People’s Republic of China

    Chengmin Huang, Peng Zhang, Tingting Li, Jun Li, Jiying Chen & Yuan Guo

  2. The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Qilu Hospital of Shandong University, Jinan, 250012, Shandong, People’s Republic of China

    Chengmin Huang, Peng Zhang, Tingting Li, Jun Li, Tianjiao Liu, Anju Zuo, Jiying Chen & Yuan Guo

  3. Department of General Medicine, Qilu Hospital of Shandong University, Jinan, 250012, Shandong, People’s Republic of China

    Tingting Li, Anju Zuo, Jiying Chen & Yuan Guo

  4. Department of Cardiology, The Affiliated Cardiovascular Hospital of Xiamen University, Xiamen, 361004, Fujian, People’s Republic of China

    Chengmin Huang

  5. Department of Geriatrics, Peking University First Hospital, Beijing, 100034, People’s Republic of China

    Peng Zhang

Authors
  1. Chengmin Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peng Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tingting Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jun Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Tianjiao Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Anju Zuo
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jiying Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yuan Guo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yuan Guo.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Huang, C., Zhang, P., Li, T. et al. Overexpression of CTRP9 attenuates the development of atherosclerosis in apolipoprotein E-deficient mice. Mol Cell Biochem 455, 99–108 (2019). https://doi.org/10.1007/s11010-018-3473-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-018-3473-y

Keywords

Search

Navigation