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CTRP3 Alleviates Ox-LDL–Induced Inflammatory Response and Endothelial Dysfunction in Mouse Aortic Endothelial Cells by Activating the PI3K/Akt/eNOS Pathway

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Abstract

C1q/tumor necrosis factor-related protein-3 (CTRP3) is a novel, certified, adipokine that beneficially regulates metabolism and inflammation in the cardiovascular system. Atherosclerotic plaque rupturing and secondary thrombosis cause vascular disorders, such as myocardial infarction and unstable angina. However, the underlying role of CTRP3 in atherosclerosis remains unclear. In this study, we aimed to elucidate whether and how CTRP3 ameliorates inflammation and endothelial dysfunction caused by oxidized low-density lipoprotein (ox-LDL). We first confirmed that CTRP3 expression was inhibited in ApoE−/− mice, compared to normal mice. Then, pcDNA-CTRP3 and siCTRP3 were transfected into mouse aortic endothelial cells after ox-LDL stimulation, and we observed that enhanced CTRP3 remarkably downregulated CRP, TNF-α, IL-6, CD40, and CD40L. We also observed that overexpression of CTRP3 elevated cell activity and decreased lactated hydrogenase release, accompanied by a marked reduction in cell apoptosis induced by ox-LDL. Meanwhile, overexpressed CTRP3 caused a decrease in Ang II, ICAM-1, and VCAM-1 expression, and it restored the balance between ET-1 and NO. Mechanism analysis confirmed that incremental CTRP3 upregulated p-PI3K, p-Akt, and p-eNOS expression, indicating that CTRP3 facilitated activation of the PI3K/Akt/eNOS pathway. On the contrary, siCTRP3 exerted the opposite effect to this activation. Blocking these pathways using LY294002 or L-NAME attenuated the protective role of CTRP3. Overall, these results suggest that CTRP3 can efficiently inhibit the inflammatory response and endothelial dysfunction induced by ox-LDL in mouse aortic endothelial cells, perhaps by activating the PI3K/Akt/eNOS pathway, indicating a promising strategy against atherosclerosis.

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References

  1. Guo, Y., X.C. Liu, Y.J. Wang, Q. Li, Q. Yang, X.G. Weng, Y. Chen, W.Y. Cai, X.X. Kan, and X. Chen. 2016. Effects of Shenlian extract on experimental atherosclerosis in ApoE-deficient mice based on ultrasound biomicroscopy. Bmc Complementary & Alternative Medicine 16: 469.

    Article  CAS  Google Scholar 

  2. Badimon, L., and G. Vilahur. 2014. Thrombosis formation on atherosclerotic lesions and plaque rupture. Journal of Internal Medicine 276: 618–632.

    Article  CAS  PubMed  Google Scholar 

  3. Ragino, Y.I., A.M. Chernjavski, Y.V. Polonskaya, A.M. Volkov, E.V. Kashtanova, A.V. Tikhonov, and S.Y. Tcimbal. 2012. Oxidation and endothelial dysfunction biomarkers of atherosclerotic plaque instability. Studies of the vascular wall and blood. Bulletin of Experimental Biology & Medicine 153: 331–335.

    Article  CAS  Google Scholar 

  4. Chong, D.P.H., and B.S. Bachenheimer. 2000. Current, new and future treatments in Dyslipidaemia and atherosclerosis. Drugs 60: 55–93.

    Article  CAS  PubMed  Google Scholar 

  5. Yoo, H.J., S.Y. Hwang, H.C. Hong, H.Y. Choi, S.J. Yang, S.C. Dong, S.H. Baik, M. Blüher, B.S. Youn, and K.M. Choi. 2013. Implication of progranulin and C1q/TNF-related protein-3 (CTRP3) on inflammation and atherosclerosis in subjects with or without metabolic syndrome. PLoS One 8: e55744.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Kim, J.Y., J.Y. Min, J.M. Baek, S.J. Ahn, H.Y. Jun, K.H. Yoon, M.K. Choi, M.S. Lee, and J. Oh. 2015. CTRP3 acts as a negative regulator of osteoclastogenesis through AMPK-c-Fos-NFATc1 signaling in vitro and RANKL-induced calvarial bone destruction in vivo. Bone 79: 242–251.

    Article  CAS  PubMed  Google Scholar 

  7. Sun, H., X. Zhu, Y. Zhou, W. Cai, and L. Qiu. 2017. C1q/TNF-related Protein-9 ameliorates ox-LDL-induced endothelial dysfunction via PGC-1α/AMPK-mediated antioxidant enzyme induction. International Journal of Molecular Sciences 18: 1097.

    Article  CAS  PubMed Central  Google Scholar 

  8. Yi, W., Y. Sun, Y. Yuan, W.B. Lau, Q. Zheng, X. Wang, Y. Wang, X. Shang, E. Gao, W.J. Koch, and X.L. Ma. 2012. C1q/tumor necrosis factor-related protein-3, a newly identified adipokine, is a novel antiapoptotic, proangiogenic, and cardioprotective molecule in the ischemic mouse heart. Circulation 125: 3159–3169.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Hofmann, C., N. Chen, F. Obermeier, G. Paul, C. Büchler, A. Kopp, W. Falk, and A. Schäffler. 2011. C1q/TNF-related Protein-3 (CTRP-3) is secreted by visceral adipose tissue and exerts antiinflammatory and antifibrotic effects in primary human colonic fibroblasts. Inflammatory Bowel Diseases 17: 2462–2471.

    Article  PubMed  Google Scholar 

  10. Schmid, A., A. Kopp, F. Hanses, T. Karrasch, and A. Schäffler. 2014. C1q/TNF-related protein-3 (CTRP-3) attenuates lipopolysaccharide (LPS)-induced systemic inflammation and adipose tissue Erk-1/-2 phosphorylation in mice in vivo. Biochemical & Biophysical Research Communications 452: 8–13.

    Article  CAS  Google Scholar 

  11. Wang, Y., L. Tu, Y. Li, D. Chen, and S. Wang. 2016. Notoginsenoside R1 protects against neonatal cerebral hypoxic-ischemic injury through estrogen receptor-dependent activation of endoplasmic reticulum stress pathways. The Journal of Pharmacology and Experimental Therapeutics 357: 591–605.

    Article  CAS  PubMed  Google Scholar 

  12. Pende, A., A. Denegri. 2012. An anti-inflammatory approach in the therapeutic choices for the prevention of atherosclerotic events: InTech, 158–164.

  13. Chang, Y., Q. Wu, T. Tian, L. Li, X. Guo, Z. Feng, J. Zhou, L. Zhang, S. Zhou, and G. Feng. 2015. The influence of SRPK1 on glioma apoptosis, metastasis, and angiogenesis through the PI3K/Akt signaling pathway under normoxia. Tumour Biology the Journal of the International Society for Oncodevelopmental Biology & Medicine 36: 6083–6093.

    Article  CAS  Google Scholar 

  14. Ou, H.C., W.J. Lee, S.D. Lee, C.Y. Huang, T.H. Chiu, K.L. Tsai, W.C. Hsu, and W.H. Sheu. 2010. Ellagic acid protects endothelial cells from oxidized low-density lipoprotein-induced apoptosis by modulating the PI3K/Akt/eNOS pathway. Toxicology & Applied Pharmacology 248: 134–143.

    Article  CAS  Google Scholar 

  15. Wygrecka, M., D. Zakrzewicz, B. Taborski, M. Didiasova, G. Kwapiszewska, K.T. Preissner, and P. Markart. 2012. TGF-Î21 induces tissue factor expression in human lung fibroblasts in a PI3K/JNK/Akt-dependent and AP-1-dependent manner. American Journal of Respiratory Cell & Molecular Biology 47: 614–627.

    Article  CAS  Google Scholar 

  16. Xing, S.S., X.Y. Yang, T. Zheng, W.J. Li, D. Wu, J.Y. Chi, F. Bian, X.L. Bai, G.J. Wu, and Y.Z. Zhang. 2015. Salidroside improves endothelial function and alleviates atherosclerosis by activating a mitochondria-related AMPK/PI3K/Akt/eNOS pathway. 中国药理学通报 72: 141.

    CAS  Google Scholar 

  17. Li, T., D. Li, H. Xu, H. Zhang, D. Tang, and H. Cao. 2016. Wen-Xin decoction ameliorates vascular endothelium dysfunction via the PI3K/AKT/eNOS pathway in experimental atherosclerosis in rats. BMC Complementary and Alternative Medicine 16: 27.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Dong, R., W. Chen, W. Feng, C. Xia, D. Hu, Y. Zhang, Y. Yang, D.W. Wang, X. Xu, and L. Tu. 2015. Exogenous bradykinin inhibits tissue factor induction and deep vein thrombosis via activating the eNOS/phosphoinositide 3-kinase/Akt signaling pathway. Cellular Physiology and Biochemistry: International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology 37: 1592–1606.

    Article  CAS  Google Scholar 

  19. Deng, G., Y. Long, Y.R. Yu, and M.R. Li. 2010. Adiponectin directly improves endothelial dysfunction in obese rats through the AMPK-eNOS pathway. International Journal of Obesity 34: 165–171.

    Article  CAS  PubMed  Google Scholar 

  20. Feng, X., Y. Zhang, R. Xu, X. Xie, L. Tao, H. Gao, Y. Gao, Z. He, and H. Wang. 2010. Lipopolysaccharide up-regulates the expression of Fcalpha/mu receptor and promotes the binding of oxidized low-density lipoprotein and its IgM antibody complex to activated human macrophages. Atherosclerosis 208: 396–405.

    Article  CAS  PubMed  Google Scholar 

  21. Matsuura, E., F. Atzeni, P. Sarzi-Puttini, M. Turiel, L.R. Lopez, and M.T. Nurmohamed. 2014. Is atherosclerosis an autoimmune disease? BMC Medicine 12: 47.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Mitra, S., A. Deshmukh, R. Sachdeva, J. Lu, and J.L. Mehta. 2011. Oxidized low-density lipoprotein and atherosclerosis implications in antioxidant therapy. The American Journal of the Medical Sciences 342: 135–142.

    Article  PubMed  Google Scholar 

  23. Huang, Y., G. Wan, and J. Tao. 2017. C1q/TNF-related protein-3 exerts the chondroprotective effects in IL-1beta-treated SW1353 cells by regulating the FGFR1 signaling. Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie 85: 41–46.

    Article  CAS  Google Scholar 

  24. Otani, M., S. Furukawa, S. Wakisaka, and T. Maeda. 2015. A novel adipokine C1q/TNF-related protein 3 is expressed in developing skeletal muscle and controls myoblast proliferation and differentiation. Molecular and Cellular Biochemistry 409: 271–282.

    Article  CAS  PubMed  Google Scholar 

  25. Zhou, B.Y., Y.L. Guo, N.Q. Wu, C.G. Zhu, Y. Gao, P. Qing, X.L. Li, Y. Wang, Q. Dong, G. Liu, R.X. Xu, C.J. Cui, J. Sun, and J.J. Li. 2017. Plasma big endothelin-1 levels at admission and future cardiovascular outcomes: A cohort study in patients with stable coronary artery disease. International Journal of Cardiology 230: 76–79.

    Article  PubMed  Google Scholar 

  26. Jr, W.T. 2007. Coronary atherosclerosis, low-density lipoproteins and markers of thrombosis, inflammation and endothelial dysfunction. International Journal of Angiology 16: 12.

    Article  Google Scholar 

  27. Hua, S., C. Song, C.L. Geczy, S.B. Freedman, and P.K. Witting. 2009. A role for acute-phase serum amyloid a and high-density lipoprotein in oxidative stress, endothelial dysfunction and atherosclerosis. Redox Report Communications in Free Radical Research 14: 187–196.

    Article  CAS  PubMed  Google Scholar 

  28. Tedgui, A. 2011. The role of inflammation in atherothrombosis: Implications for clinical practice. Vascular Medicine 10: 45–53.

    Article  Google Scholar 

  29. Kopp, A., M. Bala, C. Buechler, W. Falk, P. Gross, M. Neumeier, J. Schölmerich, and A. Schäffler. 2010. C1q/TNF-related protein-3 represents a novel and endogenous lipopolysaccharide antagonist of the adipose tissue. Endocrinology 151: 5267–5278.

    Article  CAS  PubMed  Google Scholar 

  30. Akiyama, H., S. Furukawa, S. Wakisaka, and T. Maeda. 2007. CTRP3/cartducin promotes proliferation and migration of endothelial cells. Molecular and Cellular Biochemistry 304: 243–248.

    Article  CAS  PubMed  Google Scholar 

  31. Mitra, S., T. Goyal, and J.L. Mehta. 2011. Oxidized LDL, LOX-1 and atherosclerosis. Cardiovascular Drugs and Therapy 25: 419–429.

    Article  CAS  PubMed  Google Scholar 

  32. Qiu, M.K., S.C. Wang, Y. Tang, C. Pan, Y. Wang, S.Q. Wang, Z.W. Quan, and J.M. Ou. 2017. Tim-3 inhibits low-density lipoprotein-induced atherogenic responses in human umbilical vein endothelial cells. Oncotarget 8: 61001–61010.

    PubMed  PubMed Central  Google Scholar 

  33. Nakashima, Y., E.W. Raines, A.S. Plump, J.L. Breslow, and R. Ross. 1998. Upregulation of VCAM-1 and ICAM-1 at atherosclerosis-prone sites on the endothelium in the ApoE-deficient mouse. Arteriosclerosis, Thrombosis, and Vascular Biology 18: 842–851.

    Article  CAS  PubMed  Google Scholar 

  34. Zhao, W., C. Wu, and X. Chen. 2016. Cryptotanshinone inhibits oxidized LDL-induced adhesion molecule expression via ROS dependent NF-kappaB pathways. Cell Adhesion & Migration 10: 248–258.

    Article  CAS  Google Scholar 

  35. Kojda, G., and D. Harrison. 1999. Interactions between NO and reactive oxygen species: Pathophysiological importance in atherosclerosis, hypertension, diabetes and heart failure. Cardiovascular Research 43: 562–571.

    Article  CAS  PubMed  Google Scholar 

  36. MA, B., D. I, and L. JH. 2015. Salt, angiotensin II, superoxide, and Endothelial Function. Comprehensive Physiology 6: 215.

    Google Scholar 

  37. Daugherty, A., M.W. Manning, and L.A. Cassis. 2000. Angiotensin II promotes atherosclerotic lesions and aneurysms in apolipoprotein E-deficient mice. The Journal of Clinical Investigation 105: 1605–1612.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Suo, J., L. Zhao, J. Wang, Z. Zhu, H. Zhang, and R. Gao. 2015. Influenza virus aggravates the ox-LDL-induced apoptosis of human endothelial cells via promoting p53 signaling. Journal of Medical Virology 87: 1113–1123.

    Article  CAS  PubMed  Google Scholar 

  39. Ma, Z.G., Y.P. Yuan, S.C. Xu, W.Y. Wei, C.R. Xu, X. Zhang, Q.Q. Wu, H.H. Liao, J. Ni, and Q.Z. Tang. 2017. CTRP3 attenuates cardiac dysfunction, inflammation, oxidative stress and cell death in diabetic cardiomyopathy in rats. Diabetologia 60: 1126–1137.

    Article  CAS  PubMed  Google Scholar 

  40. Pant, S., A. Deshmukh, G.S. Gurumurthy, N.V. Pothineni, T.E. Watts, F. Romeo, and J.L. Mehta. 2014. Inflammation and atherosclerosis--revisited. Journal of Cardiovascular Pharmacology and Therapeutics 19: 170–178.

    Article  CAS  PubMed  Google Scholar 

  41. Farmer, J.A., and G. Torre-Amione. 2002. Atherosclerosis and inflammation. Current Atherosclerosis Reports 4: 92–98.

    Article  PubMed  Google Scholar 

  42. Steinberg, D. 1997. Lewis A. Conner Memorial Lecture. Oxidative modification of LDL and atherogenesis. Circulation 95: 1062–1071.

    Article  CAS  PubMed  Google Scholar 

  43. Trpkovic, A., I. Resanovic, J. Stanimirovic, D. Radak, S.A. Mousa, D. Cenic-Milosevic, D. Jevremovic, and E.R. Isenovic. 2015. Oxidized low-density lipoprotein as a biomarker of cardiovascular diseases. Critical Reviews in Clinical Laboratory Sciences 52: 70–85.

    Article  CAS  PubMed  Google Scholar 

  44. Wang, S.S., S.W. Hu, Q.H. Zhang, A.X. Xia, Z.X. Jiang, and X.M. Chen. 2015. Mesenchymal stem cells stabilize atherosclerotic vulnerable plaque by anti-inflammatory properties. PLoS One 10: e0136026.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Schönbeck, U., G.K. Sukhova, K. Shimizu, F. Mach, and P. Libby. 2000. Inhibition of CD40 signaling limits evolution of established atherosclerosis in mice. Proceedings of the National Academy of Sciences of the United States of America 97: 7458–7463.

    Article  PubMed  PubMed Central  Google Scholar 

  46. Forstermann, U., and H. Li. 2011. Therapeutic effect of enhancing endothelial nitric oxide synthase (eNOS) expression and preventing eNOS uncoupling. British Journal of Pharmacology 164: 213–223.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Ning, W.H., and K. Zhao. 2013. Propionyl-L-carnitine induces eNOS activation and nitric oxide synthesis in endothelial cells via PI3 and Akt kinases. Vascular Pharmacology 59: 76–82.

    Article  CAS  PubMed  Google Scholar 

  48. Yao, Y., Y. Wang, Y. Zhang, and C. Liu. 2017. Klotho ameliorates oxidized low density lipoprotein (ox-LDL)-induced oxidative stress via regulating LOX-1 and PI3K/Akt/eNOS pathways. Lipids in Health and Disease 16: 77.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Garcia-Prieto, C.F., F. Hernandez-Nuno, D.D. Rio, G. Ruiz-Hurtado, I. Aranguez, M. Ruiz-Gayo, B. Somoza, and M.S. Fernandez-Alfonso. 2015. High-fat diet induces endothelial dysfunction through a down-regulation of the endothelial AMPK-PI3K-Akt-eNOS pathway. Molecular Nutrition & Food Research 59: 520–532.

    Article  CAS  Google Scholar 

  50. Xing, S.S., X.Y. Yang, T. Zheng, W.J. Li, D. Wu, J.Y. Chi, F. Bian, X.L. Bai, G.J. Wu, Y.Z. Zhang, C.T. Zhang, Y.H. Zhang, Y.S. Li, and S. Jin. 2015. Salidroside improves endothelial function and alleviates atherosclerosis by activating a mitochondria-related AMPK/PI3K/Akt/eNOS pathway. Vascular Pharmacology 72: 141–152.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Critical Care Medicine, Gansu Provincial Hospital of TCM, No. 418, Guazhou Road, Qilihe District, Lanzhou City, 730050, Gansu, People’s Republic of China

    Lei Chen

  2. Department of Cardiology, Lanzhou University Second Hospital, Lanzhou, 730030, Gansu, People’s Republic of China

    Lijun Qin

  3. Department of Rheumatic Osteopathology, Gansu Provincial Hospital of TCM, Lanzhou, 730050, Gansu, People’s Republic of China

    Xin Liu

  4. Central Laboratory, Gansu Provincial Hospital of TCM, Lanzhou, 730050, Gansu, People’s Republic of China

    Xiangyun Meng

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  1. Lei Chen
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Chen, L., Qin, L., Liu, X. et al. CTRP3 Alleviates Ox-LDL–Induced Inflammatory Response and Endothelial Dysfunction in Mouse Aortic Endothelial Cells by Activating the PI3K/Akt/eNOS Pathway. Inflammation 42, 1350–1359 (2019). https://doi.org/10.1007/s10753-019-00996-1

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