臺灣博碩士論文加值系統

背景: 骨髓中釋放的內皮前驅細胞在修復內皮的機制中扮演非常重要的角色，內皮前驅細胞可以移動到內皮受損的位置進行修復，以維持內皮功能的完整性。FGF21全名為fibroblast growth factor 21，主要由肝臟所分泌，具有調節能量平衡、葡萄糖與脂質代謝以及增加胰島素的敏感性等功能。近年來根據研究顯示FGF21具有對抗粥狀動脈硬化等心血管疾病的功能，但目前對於FGF21是否會影響內皮前驅細胞的功能以及內皮功能直到現在仍尚未清楚。
研究目的: 因此此篇研究主要探討FGF21是否有利於提升內皮前驅細胞的功能及釐清其機轉
結果: 根據細胞實驗結果顯示內皮前驅細胞確實有表現幫助FGF21發揮功能的β-klotho蛋白。當內皮前驅細胞受到氧化壓力的傷害下，預先給予FGF21顯著的提升內皮前驅細胞的細胞存活率且有助於恢復eNOS的活性，增加eNOS蛋白磷酸化，提升內皮前驅細胞產生一氧化氮的能力，進而改善氧化壓力抑制內皮前驅細胞功能。另一方面，預先以eNOS抑制劑L-NAME處理後， FGF21改善受氧化壓力傷害的內皮前驅細胞的好處即被抑制。在動物實驗部份我們發現持續給予ApoE-KO小鼠FGF21有助於降低血液中的膽固醇與血糖，且提升經由乙醯膽鹼刺激的血管舒張能力。另一方面給予ApoE-KO小鼠FGF21也有助於提高eNOS與SOD mRNA的表現量。相反的，持續給予健康野生型小鼠FGF21並不影響eNOS與SOD mRNA的表現量。
結論: 綜合以上結果，我們的研究初步顯示當內皮前驅細胞受氧化壓力傷害後會使eNOS表現量降低，減少一氧化氮的產生進而減弱內皮前驅細胞移動與生長的能力。而預先給予FGF21可以藉由活化Akt/eNOS/NO的訊息傳遞路徑，減少氧化壓力的傷害，有助於恢復內皮前驅細胞的功能。

Background: Circulating endothelial progenitor cells (EPCs), which function in vascular repair, represent a marker of endothelial dysfunction and vascular health. Fibroblast growth factor 21 (FGF21), a liver-secreted protein, which plays a crucial role in glucose homeostasis and lipid metabolism. Recently, FGF21 has been reported to attenuate the progression of atherosclerosis, but its impact on EPC and endothelial function remains unclear.
Aim: We hypothesized that FGF21 could ameliorate EPCs function and improve endothelial function.
Results: In the in vitro studies, the β-klotho protein was expressed in cultured EPCs. While exposed to H2O2, EPCs pre-treated with FGF21 reversed the cell viability, attenuated the H2O2-induced suppression of endothelial nitric oxide synthase (eNOS) phosphorylation, and were with higher eNOS protein levels. Additionally, FGF21 also improved the migration and tube formation functions of EPCs. However, administration of N(ω)-nitro-L-arginine methyl ester (L-NAME) inhibited FGF21’s effect of reversing oxidative injury in the EPCs. In the in vivo study, administration of FGF21 reduced the total cholesterol and blood glucose in ApoE deficient mice but no different in wild-type mice. Acetylcholine stimulated vascular relaxation were improved after treating FGF21 in ApoE deficient and wild-type mice. Moreover, FGF21 improved the mRNA expression of eNOS and antioxidant gene expression of SOD1 and SOD2 in ApoE-deficient mice. In contrast, administration of FGF21 did not impact on eNOS, SODs mRNA expression in wild-type mice.
Conclusion: The current findings suggested that FGF21 may ameliorate the H2O2-induced cellular injury and improve the EPCs functions via the Akt/eNOS/NO pathway.

誌謝 i
Contents iii
English Abstract v
中文摘要 vii
Abbreviations viii
1. Introduction 1
1.1. Cardiovascular Disease 1
1.2. Atherosclerosis 1
1.3. Endothelial progenitor cells 3
1.4. Fibroblast growth factor 21 4
1.5. Hypothesis and Study Aim 6
2. Materials and Methods 7
2.1. In vitro study
2.1.1. Human EPCs isolation and cultivation 7
2.1.2. Cell viability assay 7
2.1.3. Western bolt 8
2.1.4. Immunofluorescence staining 8
2.1.5. Measurement of Nitric Oxide production 9
2.1.6. EPCs Tube Formation Assay 9
2.1.7. EPCs Migration Assay 10
2.1.8. Measurement of ROS production 10
2.2. In vivo study
2.2.1. Animal model 11
2.2.2. Measurement of blood parameter 12
2.2.3. Measurement of vascular reactivity 12
2.2.4. Measurement of circulation EPCs levels 13
2.2.5. RNA extraction and Quantitative real-time reverse-transcription PCR 13
2.3. Statistical analysis 14
3. Results 15
3.1. Characterization of Human EPCs 15
3.2. β-klotho is expressed in EPCs 15
3.3. FGF21 ameliorates H2O2-induced cell damage in EPCs 15
3.4. FGF21 rescues H2O2-suppressed p-eNOS, p-Akt protein levels and NO production in EPCs 16
3.5. FGF21 attenuated H2O2-induced ROS production in EPCs 17
3.6. FGF21 prevents H2O2-induced inhibition of EPCs tube formation and migration 17
3.7. Effect of FGF21 on the serum lipid profiles and blood glucose levels in wild type and ApoE-KO mice 18
3.8. Effect of FGF21 on endothelial function in Wild-type and ApoE-KO mice 19
3.9. Effect of FGF21 on the mRNA expression of eNOS, SOD1 and SOD2 in wild type and ApoE-KO mice 19
3.10. Effect of FGF21 on endothelial progenitor cells in wild type and ApoE-KO mice 20
4. Discussion 21
5. Conclusion 25
6. References 26
7. Tables 34
8. Figures 36

1. Mensah GA, Brown DW. An overview of cardiovascular disease burden in the united states. Health Aff (Millwood). 2007;26:38-48
2. Knapton M, Lewin RJ, Doherty PJ. Long-term cardiovascular conditions: The role of primary care. Br J Gen Pract. 2011;61:659-660
3. Allender S, Scarborough P, Peto V, Rayner M, Leal J, Luengo-Fernandez R, Gray A. European cardiovascular disease statistics. 2008
4. Lopez AD, Mathers CD, Ezzati M, Jamison DT, Murray CJ. Global and regional burden of disease and risk factors, 2001: Systematic analysis of population health data. Lancet. 2006;367:1747-1757
5. Murray CJ, Lopez AD, Organization WH. The global burden of disease: A comprehensive assessment of mortality and disability from diseases, injuries, and risk factors in 1990 and projected to 2020: Summary. 1996
6. Ross R. Atherosclerosis--an inflammatory disease. N Engl J Med. 1999;340:115-126
7. Lusis AJ. Atherosclerosis. Nature. 2000;407:233-241
8. Davignon J, Ganz P. Role of endothelial dysfunction in atherosclerosis. Circulation. 2004;109:Iii27-32
9. Kinlay S, Libby P, Ganz P. Endothelial function and coronary artery disease. Curr Opin Lipidol. 2001;12:383-389
10. Amberger A, Maczek C, Jurgens G, Michaelis D, Schett G, Trieb K, Eberl T, Jindal S, Xu Q, Wick G. Co-expression of icam-1, vcam-1, elam-1 and hsp60 in human arterial and venous endothelial cells in response to cytokines and oxidized low-density lipoproteins. Cell Stress Chaperones. 1997;2:94-103
11. Gebuhrer V, Murphy JF, Bordet JC, Reck MP, McGregor JL. Oxidized low-density lipoprotein induces the expression of p-selectin (gmp140/padgem/cd62) on human endothelial cells. Biochem J. 1995;306 ( Pt 1):293-298
12. Sekiya M, Osuga J, Igarashi M, Okazaki H, Ishibashi S. The role of neutral cholesterol ester hydrolysis in macrophage foam cells. J Atheroscler Thromb. 2011;18:359-364
13. Moore KJ, Tabas I. Macrophages in the pathogenesis of atherosclerosis. Cell. 2011;145:341-355
14. Raines EW. The extracellular matrix can regulate vascular cell migration, proliferation, and survival: Relationships to vascular disease. Int J Exp Pathol. 2000;81:173-182
15. Newby AC, Zaltsman AB. Fibrous cap formation or destruction — the critical importance of vascular smooth muscle cell proliferation, migration and matrix formation. Cardiovasc Res. 1999;41:345-360
16. Badimon L, Vilahur G. Thrombosis formation on atherosclerotic lesions and plaque rupture. J Intern Med. 2014;276:618-632
17. Arroyo LH, Lee RT. Mechanisms of plaque rupturemechanical and biologic interactions. Cardiovasc Res. 1999;41:369-375
18. Asahara T, Masuda H, Takahashi T, Kalka C, Pastore C, Silver M, Kearne M, Magner M, Isner JM. Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization. Circul Res. 1999;85:221-228
19. Kawamoto A, Gwon HC, Iwaguro H, Yamaguchi JI, Uchida S, Masuda H, Silver M, Ma H, Kearney M, Isner JM, Asahara T. Therapeutic potential of ex vivo expanded endothelial progenitor cells for myocardial ischemia. Circulation. 2001;103:634-637
20. Asahara T, Murohara T, Sullivan A, Silver M, van der Zee R, Li T, Witzenbichler B, Schatteman G, Isner JM. Isolation of putative progenitor endothelial cells for angiogenesis. Science. 1997;275:964-967
21. Minhajat R, Nilasari D, Bakri S. The role of endothelial progenitor cell in cardiovascular disease risk factors. Acta Med Indones. 2015;47:340-347
22. Khan SS, Solomon MA, McCoy JP, Jr. Detection of circulating endothelial cells and endothelial progenitor cells by flow cytometry. Cytometry B Clin Cytom. 2005;64:1-8
23. Yoder MC. Defining human endothelial progenitor cells. J Thromb Haemost. 2009;7 Suppl 1:49-52
24. Mannarino E, Pirro M. Endothelial injury and repair: A novel theory for atherosclerosis. Angiology. 2008;59:69s-72s
25. Luo Y, Zhao Y, Li X, Zhao J, Zhang W. Znf580 mediates enos expression and endothelial cell migration/proliferation via the tgf-beta1/alk5/smad2 pathway. Mol Cell Biochem. 2014;393:199-207
26. Ozuyaman B, Ebner P, Niesler U, Ziemann J, Kleinbongard P, Jax T, Godecke A, Kelm M, Kalka C. Nitric oxide differentially regulates proliferation and mobilization of endothelial progenitor cells but not of hematopoietic stem cells. Thromb Haemost. 2005;94:770-772
27. Vasa M, Fichtlscherer S, Aicher A, Adler K, Urbich C, Martin H, Zeiher AM, Dimmeler S. Number and migratory activity of circulating endothelial progenitor cells inversely correlate with risk factors for coronary artery disease. Circ Res. 2001;89:E1-7
28. Tepper OM, Galiano RD, Capla JM, Kalka C, Gagne PJ, Jacobowitz GR, Levine JP, Gurtner GC. Human endothelial progenitor cells from type ii diabetics exhibit impaired proliferation, adhesion, and incorporation into vascular structures. Circulation. 2002;106:2781-2786
29. Giannotti G, Doerries C, Mocharla PS, Mueller MF, Bahlmann FH, Horvath T, Jiang H, Sorrentino SA, Steenken N, Manes C, Marzilli M, Rudolph KL, Luscher TF, Drexler H, Landmesser U. Impaired endothelial repair capacity of early endothelial progenitor cells in prehypertension: Relation to endothelial dysfunction. Hypertension. 2010;55:1389-1397
30. Choi JH, Kim KL, Huh W, Kim B, Byun J, Suh W, Sung J, Jeon ES, Oh HY, Kim DK. Decreased number and impaired angiogenic function of endothelial progenitor cells in patients with chronic renal failure. Arterioscler Thromb Vasc Biol. 2004;24:1246-1252
31. Lee SH, Kim JY, Yoo SY, Kwon SM. Cytoprotective effect of dieckol on human endothelial progenitor cells (hepcs) from oxidative stress-induced apoptosis. Free Radic Res. 2013;47:526-534
32. Imanishi T, Hano T, Sawamura T, Nishio I. Oxidized low-density lipoprotein induces endothelial progenitor cell senescence, leading to cellular dysfunction. Clin Exp Pharmacol Physiol. 2004;31:407-413
33. Dernbach E, Urbich C, Brandes RP, Hofmann WK, Zeiher AM, Dimmeler S. Antioxidative stress-associated genes in circulating progenitor cells: Evidence for enhanced resistance against oxidative stress. Blood. 2004;104:3591-3597
34. Cheng P, Zhang F, Yu L, Lin X, He L, Li X, Lu X, Yan X, Tan Y, Zhang C. Physiological and pharmacological roles of fgf21 in cardiovascular diseases. J Diabetes Res. 2016;2016:1540267
35. Itoh N, Ornitz DM. Evolution of the fgf and fgfr gene families. Trends Genet. 2004;20:563-569
36. Nishimura T, Nakatake Y, Konishi M, Itoh N. Identification of a novel fgf, fgf-21, preferentially expressed in the liver. Biochim Biophys Acta. 2000;1492:203-206
37. Suzuki M, Uehara Y, Motomura-Matsuzaka K, Oki J, Koyama Y, Kimura M, Asada M, Komi-Kuramochi A, Oka S, Imamura T. Betaklotho is required for fibroblast growth factor (fgf) 21 signaling through fgf receptor (fgfr) 1c and fgfr3c. Mol Endocrinol. 2008;22:1006-1014
38. Goetz R, Beenken A, Ibrahimi OA, Kalinina J, Olsen SK, Eliseenkova AV, Xu C, Neubert TA, Zhang F, Linhardt RJ, Yu X, White KE, Inagaki T, Kliewer SA, Yamamoto M, Kurosu H, Ogawa Y, Kuro-o M, Lanske B, Razzaque MS, Mohammadi M. Molecular insights into the klotho-dependent, endocrine mode of action of fibroblast growth factor 19 subfamily members. Mol Cell Biol. 2007;27:3417-3428
39. Hondares E, Iglesias R, Giralt A, Gonzalez FJ, Giralt M, Mampel T, Villarroya F. Thermogenic activation induces fgf21 expression and release in brown adipose tissue. J Biol Chem. 2011;286:12983-12990
40. Wente W, Efanov AM, Brenner M, Kharitonenkov A, Koster A, Sandusky GE, Sewing S, Treinies I, Zitzer H, Gromada J. Fibroblast growth factor-21 improves pancreatic beta-cell function and survival by activation of extracellular signal-regulated kinase 1/2 and akt signaling pathways. Diabetes. 2006;55:2470-2478
41. Izumiya Y, Bina HA, Ouchi N, Akasaki Y, Kharitonenkov A, Walsh K. Fgf21 is an akt-regulated myokine. FEBS Lett. 2008;582:3805-3810
42. Bookout AL, de Groot MHM, Owen BM, Lee S, Gautron L, Lawrence HL, Ding X, Elmquist JK, Takahashi JS, Mangelsdorf DJ, Kliewer SA. Fgf21 regulates metabolism and circadian behavior by acting on the nervous system. Nat Med. 2013;19:1147
43. Planavila A, Redondo-Angulo I, Ribas F, Garrabou G, Casademont J, Giralt M, Villarroya F. Fibroblast growth factor 21 protects the heart from oxidative stress. Cardiovasc Res. 2015;106:19-31
44. Kharitonenkov A, Shiyanova TL, Koester A, Ford AM, Micanovic R, Galbreath EJ, Sandusky GE, Hammond LJ, Moyers JS, Owens RA, Gromada J, Brozinick JT, Hawkins ED, Wroblewski VJ, Li DS, Mehrbod F, Jaskunas SR, Shanafelt AB. Fgf-21 as a novel metabolic regulator. J Clin Invest. 2005;115:1627-1635
45. Hondares E, Rosell M, Gonzalez FJ, Giralt M, Iglesias R, Villarroya F. Hepatic fgf21 expression is induced at birth via pparalpha in response to milk intake and contributes to thermogenic activation of neonatal brown fat. Cell Metab. 2010;11:206-212
46. Fisher FM, Kleiner S, Douris N, Fox EC, Mepani RJ, Verdeguer F, Wu J, Kharitonenkov A, Flier JS, Maratos-Flier E, Spiegelman BM. Fgf21 regulates pgc-1alpha and browning of white adipose tissues in adaptive thermogenesis. Genes Dev. 2012;26:271-281
47. Mraz M, Bartlova M, Lacinova Z, Michalsky D, Kasalicky M, Haluzikova D, Matoulek M, Dostalova I, Humenanska V, Haluzik M. Serum concentrations and tissue expression of a novel endocrine regulator fibroblast growth factor-21 in patients with type 2 diabetes and obesity. Clin Endocrinol (Oxf). 2009;71:369-375
48. Li H, Bao Y, Xu A, Pan X, Lu J, Wu H, Lu H, Xiang K, Jia W. Serum fibroblast growth factor 21 is associated with adverse lipid profiles and gamma-glutamyltransferase but not insulin sensitivity in chinese subjects. J Clin Endocrinol Metab. 2009;94:2151-2156
49. Bobbert T, Schwarz F, Fischer-Rosinsky A, Pfeiffer AF, Mohlig M, Mai K, Spranger J. Fibroblast growth factor 21 predicts the metabolic syndrome and type 2 diabetes in caucasians. Diabetes Care. 2013;36:145-149
50. Stein S, Bachmann A, Lossner U, Kratzsch J, Bluher M, Stumvoll M, Fasshauer M. Serum levels of the adipokine fgf21 depend on renal function. Diabetes Care. 2009;32:126-128
51. Planavila A, Redondo-Angulo I, Villarroya F. Fgf21 and cardiac physiopathology. Front Endocrinol (Lausanne). 2015;6
52. Planavila A, Redondo I, Hondares E, Vinciguerra M, Munts C, Iglesias R, Gabrielli LA, Sitges M, Giralt M, van Bilsen M, Villarroya F. Fibroblast growth factor 21 protects against cardiac hypertrophy in mice. Nat Commun. 2013;4:2019
53. Lin Z, Pan X, Wu F, Ye D, Zhang Y, Wang Y, Jin L, Lian Q, Huang Y, Ding H, Triggle C, Wang K, Li X, Xu A. Fibroblast growth factor 21 prevents atherosclerosis by suppression of hepatic sterol regulatory element-binding protein-2 and induction of adiponectin in mice. Circulation. 2015;131:1861-1871
54. Zhu W, Wang C, Liu L, Li Y, Li X, Cai J, Wang H. Effects of fibroblast growth factor 21 on cell damage in vitro and atherosclerosis in vivo. Can J Physiol Pharmacol. 2014;92:927-935
55. Werner N, Wassmann S, Ahlers P, Schiegl T, Kosiol S, Link A, Walenta K, Nickenig G. Endothelial progenitor cells correlate with endothelial function in patients with coronary artery disease. Basic Res Cardiol. 2007;102:565-571
56. Boilson BA, Kiernan TJ, Harbuzariu A, Nelson RE, Lerman A, Simari RD. Circulating cd34+ cell subsets in patients with coronary endothelial dysfunction. Nat Clin Pract Cardiovasc Med. 2008;5:489-496
57. Chen JZ, Zhu JH, Wang XX, Zhu JH, Xie XD, Sun J, Shang YP, Guo XG, Dai HM, Hu SJ. Effects of homocysteine on number and activity of endothelial progenitor cells from peripheral blood. J Mol Cell Cardiol. 2004;36:233-239
58. Huang PH, Chen YH, Wang CH, Chen JS, Tsai HY, Lin FY, Lo WY, Wu TC, Sata M, Chen JW, Lin SJ. Matrix metalloproteinase-9 is essential for ischemia-induced neovascularization by modulating bone marrow-derived endothelial progenitor cells. Arterioscler Thromb Vasc Biol. 2009;29:1179-1184
59. Hennessey JC, McGuire JJ. Attenuated vasodilator effectiveness of protease-activated receptor 2 agonist in heterozygous par2 knockout mice. PLoS One. 2013;8:e55965
60. Kurosu H, Choi M, Ogawa Y, Dickson AS, Goetz R, Eliseenkova AV, Mohammadi M, Rosenblatt KP, Kliewer SA, Kuro-o M. Tissue-specific expression of betaklotho and fibroblast growth factor (fgf) receptor isoforms determines metabolic activity of fgf19 and fgf21. J Biol Chem. 2007;282:26687-26695
61. Ogawa Y, Kurosu H, Yamamoto M, Nandi A, Rosenblatt KP, Goetz R, Eliseenkova AV, Mohammadi M, Kuro-o M. Betaklotho is required for metabolic activity of fibroblast growth factor 21. Proc Natl Acad Sci U S A. 2007;104:7432-7437
62. Deanfield JE, Halcox JP, Rabelink TJ. Endothelial function and dysfunction: Testing and clinical relevance. Circulation. 2007;115:1285-1295
63. Tousoulis D, Psaltopoulou T, Androulakis E, Papageorgiou N, Papaioannou S, Oikonomou E, Synetos A, Stefanadis C. Oxidative stress and early atherosclerosis: Novel antioxidant treatment. Cardiovasc Drugs Ther. 2015;29:75-88
64. Chen X, Chen Q, Wang L, Li G. Ghrelin induces cell migration through ghsr1a-mediated pi3k/akt/enos/no signaling pathway in endothelial progenitor cells. Metabolism. 2013;62:743-752
65. Everaert BR, Van Craenenbroeck EM, Hoymans VY, Haine SE, Van Nassauw L, Conraads VM, Timmermans JP, Vrints CJ. Current perspective of pathophysiological and interventional effects on endothelial progenitor cell biology: Focus on pi3k/akt/enos pathway. Int J Cardiol. 2010;144:350-366
66. Yu X, Song M, Chen J, Zhu G, Zhao G, Wang H, Hunag L. Hepatocyte growth factor protects endothelial progenitor cell from damage of low-density lipoprotein cholesterol via the pi3k/akt signaling pathway. Mol Biol Rep. 2010;37:2423-2429
67. Li Y, Huang J, Jiang Z, Jiao Y, Wang H. Fgf21 inhibitor suppresses the proliferation and migration of human umbilical vein endothelial cells through the enos/pi3k/akt pathway. Am J Transl Res. 2017;9:5299-5307
68. Fulton D, Gratton J-P, McCabe TJ, Fontana J, Fujio Y, Walsh K, Franke TF, Papapetropoulos A, Sessa WC. Regulation of endothelium-derived nitric oxide production by the protein kinase akt. Nature. 1999;399:597
69. Li H, Forstermann U. Uncoupling of endothelial no synthase in atherosclerosis and vascular disease. Curr Opin Pharmacol. 2013;13:161-167
70. Li H, Horke S, Forstermann U. Vascular oxidative stress, nitric oxide and atherosclerosis. Atherosclerosis. 2014;237:208-219
71. Bookout AL, de Groot MH, Owen BM, Lee S, Gautron L, Lawrence HL, Ding X, Elmquist JK, Takahashi JS, Mangelsdorf DJ, Kliewer SA. Fgf21 regulates metabolism and circadian behavior by acting on the nervous system. Nat Med. 2013;19:1147-1152
72. Wang XM, Song SS, Xiao H, Gao P, Li XJ, Si LY. Fibroblast growth factor 21 protects against high glucose induced cellular damage and dysfunction of endothelial nitric-oxide synthase in endothelial cells. Cell Physiol Biochem. 2014;34:658-671
73. Dimmeler S, Aicher A, Vasa M, Mildner-Rihm C, Adler K, Tiemann M, Rutten H, Fichtlscherer S, Martin H, Zeiher AM. Hmg-coa reductase inhibitors (statins) increase endothelial progenitor cells via the pi 3-kinase/akt pathway. J Clin Invest. 2001;108:391-397
74. Aicher A, Heeschen C, Mildner-Rihm C, Urbich C, Ihling C, Technau-Ihling K, Zeiher AM, Dimmeler S. Essential role of endothelial nitric oxide synthase for mobilization of stem and progenitor cells. Nat Med. 2003;9:1370-1376
75. Case J, Ingram DA, Haneline LS. Oxidative stress impairs endothelial progenitor cell function. Antioxid Redox Signal. 2008;10:1895-1907
76. Yu Y, Bai F, Wang W, Liu Y, Yuan Q, Qu S, Zhang T, Tian G, Li S, Li D, Ren G. Fibroblast growth factor 21 protects mouse brain against d-galactose induced aging via suppression of oxidative stress response and advanced glycation end products formation. Pharmacol Biochem Behav. 2015;133:122-131
77. Yu Y, Bai F, Liu Y, Yang Y, Yuan Q, Zou D, Qu S, Tian G, Song L, Zhang T, Li S, Liu Y, Wang W, Ren G, Li D. Fibroblast growth factor (fgf21) protects mouse liver against d-galactose-induced oxidative stress and apoptosis via activating nrf2 and pi3k/akt pathways. Mol Cell Biochem. 2015;403:287-299
78. Yang H, Feng A, Lin S, Yu L, Lin X, Yan X, Lu X, Zhang C. Fibroblast growth factor-21 prevents diabetic cardiomyopathy via ampk-mediated antioxidation and lipid-lowering effects in the heart. Cell Death Dis. 2018;9:227
79. Forstermann U, Munzel T. Endothelial nitric oxide synthase in vascular disease: From marvel to menace. Circulation. 2006;113:1708-1714
80. Andersen B, Omar BA, Rakipovski G, Raun K, Ahren B. Fibroblast growth factor 21 prevents glycemic deterioration in insulin deficient mouse models of diabetes. Eur J Pharmacol. 2015;764:189-194
81. d'Uscio LV, Baker TA, Mantilla CB, Smith L, Weiler D, Sieck GC, Katusic ZS. Mechanism of endothelial dysfunction in apolipoprotein e-deficient mice. Arterioscler Thromb Vasc Biol. 2001;21:1017-1022
82. Faraci FM, Didion SP. Vascular protection: Superoxide dismutase isoforms in the vessel wall. Arterioscler Thromb Vasc Biol. 2004;24:1367-1373
83. Yang H, Roberts LJ, Shi MJ, Zhou LC, Ballard BR, Richardson A, Guo ZM. Retardation of atherosclerosis by overexpression of catalase or both cu/zn-superoxide dismutase and catalase in mice lacking apolipoprotein e. Circ Res. 2004;95:1075-1081