- Original investigation
- Open Access
Fibroblast growth factor 21 deletion aggravates diabetes-induced pathogenic changes in the aorta in type 1 diabetic mice
© Yan et al. 2015
- Received: 15 January 2015
- Accepted: 2 June 2015
- Published: 11 June 2015
Fibroblast growth factor 21 (FGF21) is an important regulator in glucose and lipid metabolism, and has been considered as a potential therapy for diabetes. The effect of FGF21 on the development and progression of diabetes-induced pathogenic changes in the aorta has not currently been addressed. To characterize these effects, type 1 diabetes was induced in both FGF21 knockout (FGF21KO) and C57BL/6 J wild type (WT) mice via multiple-dose streptozotocin injection. FGF21KO diabetic mice showed both earlier and more severe aortic remodeling indicated by aortic thickening, collagen accumulation and fibrotic mediator connective tissue growth factor expression. This was accompanied by significant aortic cell apoptosis than in WT diabetic mice. Further investigation found that FGF21 deletion exacerbated aortic inflammation and oxidative stress reflected by elevated expression of tumor necrosis factor α and transforming growth factor β, and the accumulation of 3-nitrotyrocine and 4-Hydroxynonenal. FGF21 administration can reverse the pathologic changes in FGF21KO diabetic mice. These findings demonstrate that FGF21 deletion aggravates aortic remodeling and cell death probably via exacerbation of aortic inflammation and oxidative stress. This marks FGF21 as a potential therapy for the treatment of aortic damage due to diabetes.
- Fibroblast growth factor 21
- Vascular damage
- Oxidative stress
Diabetic vascular complications, including macroangiopathy, microangiopathy and peripheral vascular complications, are the most common diabetic complications in both type 1  and type 2  diabetes mellitus and make major contributions to diabetic mortality and morbidity . Diabetic microvascular disease is a leading cause of blindness, renal failure and nerve damage. Furthermore, diabetic macroangiopathy and peripheral vascular complications lead to increased risk of myocardial infarction, stroke and limb amputation . About 80 % of all diabetic patients die from cardiovascular events. Of which, 75 % are due to coronary heart disease and the remaining 25 % are attributed to cerebrovascular, peripheral or other macrovascular disease .
Even though the exact mechanism for accelerated vascular disease in diabetes is not yet fully clear, existing research has defined numerous risk factors involved in diabetes, such as oxidative stress [5, 6], dyslipidemia [4, 5], advanced glycation , decline in nitric oxide production, activation of the renin-angiotensin aldosterone system, and endothelial inflammation . All contribute to the development of diabetic vascular complications.
Fibroblast growth factor 21 (FGF21), a newly-defined member of the FGF family , has been identified as a potent metabolic regulator with specific effects on glucose and lipid metabolism . FGF21 can stimulate glucose uptake in adipocytes , and enhance glucose clearance by enhancing the browning of white adipose tissues . In response to fasting, FGF21 can regulate lipolysis in adipocytes . FGF21 also shows beneficial effects on lipid profiles as demonstrated by lower circulating lipids in both rodent  and primate  diabetic models following FGF21 administration. FGF21 treatment also enhanced expression and secretion of the downstream effector, adiponectin, in adipocytes, which in turn further improved fatty acid oxidation and lipid clearance in the liver and skeletal muscle . Moreover, FGF21 has an insulin-sensitizing ability  and can ameliorate glucose tolerance  by reducing hepatic glucose production and stimulating glucose uptake in adipocytes.
Because of its ability to regulate glucose and lipid metabolism, FGF21 has shown therapeutic potential in treating diabetes . FGF21 transgenic mice were lean and resistant to age-associated or diet-induced obesity and insulin resistance . Both acute  and chronic  administration of FGF21 can ameliorate the metabolic state of diabetes. FGF21 treatment resulted in rapid decline of blood glucose levels and immediate improvement of glucose tolerance and insulin sensitivity in both ob/ob and diet-induced obese mice [18, 19] over the short term and ameliorated fasting hyperglycemia in both ob/ob mice  and diabetic monkeys  over the long term treatment. In addition, the level of serum FGF21 is reported to be positively associated with coronary artery disease  and higher risk of cardiovascular events in patients with type 2 diabetes , which might indicate a compensatory response. However, the direct effects of FGF21 on diabetic complications still remain largely unknown.
Almost all specific risk factors of diabetic vascular complications are directly related to hyperglycemia  and/or hyperlipidemia . Ameliorating glucose and lipid metabolism is still a major preventive and assistive therapeutic strategy for diabetic vascular complications. Considering the anti-hyperglycemic and anti-hyperlipidemic effects of FGF21 on diabetes, and the fact that its preferred receptor, fibroblast growth factor receptor 1c (FGFR1c), and co-receptor, β-klotho, are highly-expressed in aorta , FGF21 is indicated to be involved in pathogenic changes in the aorta under diabetic conditions. Therefore, we investigated the role of FGF21 in the development and progression of pathogenic changes in the aorta in a streptozotocin (STZ)-induced type 1 diabetic model using FGF21 knockout (FGF21KO) mice.
This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The protocol was approved by the Animal Policy and Welfare Committee of Wenzhou Medical University and the Institutional Animal Care and Use Committee of the University of Louisville. All surgeries were performed under anesthesia induced by intraperitoneal injection of 1.2 % 2,2,2-Tribromoethanol (Avertin) at the dose of 300 mg/kg body weight and all efforts were made to minimize suffering.
The present study used male FGF21KO mice with C57 BL/6 J background (gifted by Dr. Steve Kliewer, University of Texas Southwestern Medical Center)  and wild type (WT) C57 BL/6 J mice purchased from Jackson Laboratory (Bar Harbor, Maine). The type 1 diabetes model was induced in 10 week-old male FGF21KO mice and age-matched WT mice by intraperitoneal injection of 6 consecutive doses of STZ (60 mg/kg body weight, Sigma, St. Louis, MO) in 10 mM sodium citrate buffer, pH 4.5. FGF21KO and WT mice control groups (Ctrl) received citrate buffer alone. Seven days after the last STZ injection, whole blood glucose obtained from the mouse tail vein was assayed using a SureStep complete blood glucose monitor (LifeScan, Milpitas, CA). Animals with blood glucose levels greater than 250 mg/dL were considered diabetic. At 1, 2 and 4 months following diabetes onset, mice were sacrificed and aorta tissue was collected.
In FGF21 treatment experiment, an acute type 1 diabetic model was induced in 10 week-old male FGF21KO mice and age-matched WT mice as described above. FGF21KO and WT mice control groups (Ctrl) received citrate buffer alone. FGF21KO diabetic mice in FGF21 treatment group received intraperitoneal injection of FGF21 (100 μg/kg body weight per day) for 2 months. Thereafter, mice were sacrificed and aorta tissue was collected.
Aorta sample preparation and histopathological examination
Under anesthesia, thoracotomies were performed on mice and the descending thoracic aortas were carefully harvested and fixed in 10 % buffered formalin. Next, aorta tissues were cut into ring segments (2–3 mm in length), dehydrated in graded alcohol, cleared with xylene, and finally embedded in paraffin. Sections (5 μm thickness) were cut for pathological and immunohistochemical staining. Histological changes in the aorta were evaluated by hematoxylin and eosin (H&E) staining using Image Pro Plus 6.0 software for measuring the means of the tunica media width as the thickness of aortic tunica media.
Sirius-red staining for collagen
Aortic fibrosis was evaluated by Sirius-red staining, as described previously . Briefly, 5 μm tissue sections were stained with 0.1 % Sirius-red F3BA and 0.25 % Fast Green FCF and assessed for the proportion of collagen using a Nikon Eclipse E600 microscopy system.
Terminal deoxynucleotidyl-transferase-mediated dUTP nick-end labeling (TUNEL) staining was performed on formalin-fixed, paraffin-embedded sections with Peroxidase In Situ Apoptosis Detection Kit (Millipore, Billerica, MA) according to the manufacturer’s instructions and nuclei were stained using methyl green (FD Neurotechnologies, Columbia, MD). Positively stained apoptotic cells were counted randomly in a minimum of five microscopic fields in each of the three slides per aorta under light microscopy. The percentage of TUNEL positive cells relative to 100 nuclei was presented.
Formalin-fixed, paraffin-embedded aorta sections were dewaxed using xylene and rehydrated by serial washes in graded alcohol and a final wash in dH2O for 15 min. After balanced with phosphate buffered saline (PBS), aorta sections were incubated in Target Retrieval buffer (DAKO, Carpinteria, CA) at 95 °C, and endogenous peroxidase was quenched by incubating in 3 % H2O2 at room temperature for 10 min. After washing with PBS 3 times, sections were blocked in 5 % bovine serum albumin (BSA) for 30 min, then incubated with primary antibody against mice tumor necrosis factor α (TNF-α), connective tissue growth factor (CTGF), transforming growth factor β (TGF-β), 3-nitrotyrocine (3-NT), 4-Hydroxynonenal (4-HNE), nuclear factor E2-related factor-2 (Nrf2) or phosphorylated endothelial nitric oxide synthase (p-eNOS, Ser 1177) overnight at 4 °C. Sections incubated with PBS were used as negative controls. After washing, sections were incubated with corresponding secondary antibodies at room temperature for 1 h. For the development of color, sections were treated with peroxidase substrate DAB kit (Vector Laboratories, Inc. Burlingame, CA) and counterstained with hematoxylin. Quantitative analysis was carried out using Image J software.
Enzyme linked immunosorbent assay (ELISA)
Whole blood was collected in a lithium heparin tube (BD, Franklin Lakes, NJ) and centrifuged at 2000 rpm for 20 min. Plasma was used for interleukin- 6 (IL- 6) assay using a mouse IL-6 ELISA kit (Invitrogen, Frederick, MD) according to the manufacturer’s instructions.
Data were collected from several animals (n=5 ~ 9) and presented as means ± SD. Image Pro Plus 6.0 software was used to measure pathological changes as described above. Comparisons were performed by one-way ANOVA for the different groups, followed by post hoc pairwise repetitive comparisons using Tukey’s test. Statistical analysis was done using Origin 7.5 Lab data analysis and graphing software. Statistical significance was considered as P <0.05.
FGF21 deletion accelerated diabetes-induced aortic thickening
FGF21 deletion aggravated diabetes-induced aortic fibrosis
The aggravated fibrosis was also confirmed by immunohistochemical staining for the fibrotic mediator, CTGF. It was demonstrated that diabetes significantly up-regulated CTGF expression in both WT and FGF21KO diabetic mice at 4 months compared to their corresponding controls. This was significantly higher in FGF21KO diabetic mice than in WT diabetic mice (Fig. 2c&d). However, FGF21 deletion had no significant effect on CTGF expression under non-diabetic conditions compared to WT control mice at all 3 time points.
FGF21 deletion exacerbated diabetes-induced aortic inflammation
FGF21 deletion aggravated diabetes-induced aortic cell apoptosis
FGF21 deletion exacerbated diabetes-induced aortic oxidative stress
Nrf2, a transcription factor in regulation of various antioxidative and cytoprotective responses, has been shown to play an important role in cellular prevention against oxidative stress and damage in vitro and in vivo . In FGF21KO diabetic mice, the aortic Nrf2 expression was significantly up-regulated, especially at the 4th month (Fig. 5e&f), indicating an adaptive response to the aggravated oxidative stress.
FGF21 deletion exacerbated diabetes-induced impairment of eNOS activation
FGF21 administration ameliorated diabetes induced aorta dysfunction
The therapeutic effect of FGF21 on diabetes and diabetic complications has been widely appreciated [13, 14]. But its effect on diabetic vasculopathy remains largely unknown. High level expression of the preferred receptor, FGFR1c, and co-receptor, β-klotho, in the aorta  indicates that aorta is a potential target tissue of FGF21. By using the FGF21KO mouse model, we provided the first experimental evidence to show that FGF21 deletion further accelerated and aggravated diabetes-induced aortic thickening, fibrotic remodeling, inflammation, cell apoptosis and oxidative stress, and FGF21 administration can reverse the pathologic changes in FGF21KO diabetic mice.
FGF21 has been considered as a potent regulator in glucose and lipid metabolism. The fact that blood glucose and lipid levels are comparable in WT and FGF21KO diabetic mice  and that FGF21 deletion further aggravates of the aortic thickening (Fig. 1), fibrosis (Fig. 2), inflammation (Fig. 3), cell apoptosis (Fig. 4) and oxidative stress (Fig. 5), suggests that the detrimental effect of FGF21 deletion on the aorta is most likely mediated by its direct action in aortic tissues rather than secondary actions such as manipulating systemic glucose and/or lipid metabolism.
The endothelium, which consists of a metabolically active monolayer of endothelial cells covering the entire luminal surface of blood vessels, plays a fundamental role in maintaining vascular homeostasis. Endothelial dysfunction was considered as a starting point for macroangiopathy and microangiopathy in both type1 and type 2 diabetes which would trigger the development of diabetic vasculopathy . Cell apoptosis was assessed as an initial step of endothelial dysfunction . In the present study, we found that FGF21 deletion accelerated and aggravated diabetes-induced aortic cell apoptosis (Fig. 4). These results are consistent with recent studies that FGF21 inhibits endothelial cell apoptosis induced by oxidized low density lipoprotein  or high glucose , enhances cell viability and decreases the apoptotic cell death in human umbilical vein endothelial cells (HUVECs) caused by H2O2 stress induction in vitro, while improves the condition of atherosclerotic rats in vivo [35, 36].
Chronic inflammation and oxidative stress play important roles in the development and progression of various chronic vascular pathological changes, including endothelial remodeling and apoptotic cell death under diabetic conditions . It has been shown that FGF21 plays an important protective role against alcoholic fatty liver disease , drug-induced hepatotoxicity , atherosclerosis  and diabetic nephropathy  through its anti-oxidative stress and/ or anti-inflammatory actions. Herein we found that FGF21 deletion aggravated diabetes-induced oxidative stress, inflammation and fibrotic remodeling in aortas, reflected in the exacerbated accumulation of 3-NT and 4-HNE (Fig. 5 a-d), expression of TGF-β and TNF-α (Fig. 3), and accumulation of collagen and CTGF expression (Fig. 2), respectively. These results are consistent with a previous report that FGF21 deletion markedly aggravated acetaminophen overdose-induced liver damage, which was accompanied by increased oxidative stress and impaired antioxidant capacities. The replenishment of recombinant FGF21 largely reversed acetaminophen-induced hepatic oxidative stress and liver injury in FGF21KO mice , and are concurrent with our previous studies that demonstrated FGF21 administration attenuates diabetes-induced oxidative stress and inflammation in testis  and kidney .
One possible cause for the aggravated pathological changes of the aorta in FGF21KO diabetic mice is the dysfunction of eNOS. eNOS gene deficiency resulted in hypertension , increased vascular smooth muscle cell proliferation in response to vessel injury , increased leukocyte-endothelial interactions , hypercoagulability  and increased diet-induced atherosclerosis . Recently, an in vitro study  showed that eNOS phosphorylation at Ser-1177 and Ser-633 in HUVECs was impaired under diabetic conditions, which can be rescued by FGF21 administration in an AMP-activated protein kinase-dependent manner. In present study, phosphorylation of eNOS at Ser-1177 was further down-regulated in FGF21KO diabetic mice than that in WT diabetic mice (Fig. 6), which indicated that FGF21 deficiency may contribute to the aggravated aortic damage by impairing eNOS activation.
In conclusion, we found that FGF21 deletion accelerates and aggravates diabetes-induced aortic pathological changes reflected by exacerbated aortic thickening, collagen accumulation and fibrotic remodeling, which is most likely due to FGF21 deficiency-induced aggravation of aortic oxidative stress, inflammation, and cell apoptosis, and FGF21 administration can reverse those pathologic changes in FGF21KO diabetic mice.
This study was supported in part by Research Development Fund of Wenzhou Medical University (QTJ13007, QTJ13005), a Junior Faculty Award (1-13-JF-53), the National Natural Science Foundation of China (81273509, 81200239, 81370917, 81270293, 81470061), Key Science and Technology Development Plan from Wenzhou City (Y20140735, Y20140654), a Project of Medical Technology in Zhejiang Province (201472233), a Key New Drug Development Grants (2012ZX09103-301-016), a Changjiang Innovation Team Program (2010R50042-17), and a Starting-Up Fund for Chinese-American Research Institute for Diabetic Complications (Wenzhou Medical University). The authors gratefully acknowledge Leroy R Sachleben Jr for his editorial help.
- Schnell O, Cappuccio F, Genovese S, Standl E, Valensi P, Ceriello A. Type 1 diabetes and cardiovascular disease. Cardiovasc Diabetol. 2013;12:126.View ArticleGoogle Scholar
- Laight DW, Carrier MJ, Anggard EE. Endothelial cell dysfunction and the pathogenesis of diabetic macroangiopathy. Diabetes-Metab Res. 1999;15(4):274–82.View ArticleGoogle Scholar
- Brownlee M. Biochemistry and molecular cell biology of diabetic complications. Nature. 2001;414(6865):813–20.PubMedView ArticleGoogle Scholar
- Coccheri S. Approaches to prevention of cardiovascular complications and events in diabetes mellitus. Drugs. 2007;67(7):997–1026.PubMedView ArticleGoogle Scholar
- Laight DW, Carrier MJ, Anggard EE. Antioxidants, diabetes and endothelial dysfunction. Cardiovasc Res. 2000;47(3):457–64.PubMedView ArticleGoogle Scholar
- Gartner V, Eigentler TK. Pathogenesis of diabetic macro- and microangiopathy. Clin Nephrol. 2008;70(1):1–9.PubMedView ArticleGoogle Scholar
- Gugliucci A. Glycation as the glucose link to diabetic complications. J Am Osteopath Assoc. 2000;100(10):621–34.PubMedGoogle Scholar
- Nishimura T, Nakatake Y, Konishi M, Itoh N. Identification of a novel FGF, FGF-21, preferentially expressed in the liver. Bba-Gene Struct Expr. 2000;1492(1):203–6.View ArticleGoogle Scholar
- Adams AC, Kharitonenkov A. FGF21: The center of a transcriptional nexus in metabolic regulation. Curr Diabetes Rev. 2012;8(4):285–93.PubMedView ArticleGoogle Scholar
- Iglesias P, Selgas R, Romero S, Diez JJ. Biological role, clinical significance, and therapeutic possibilities of the recently discovered metabolic hormone fibroblastic growth factor 21. Eur J Endocrinol. 2012;167(3):301–9.PubMedView ArticleGoogle Scholar
- Fisher FM, Kleiner S, Douris N, Fox EC, Mepani RJ, Verdeguer F, et al. FGF21 regulates PGC-1 alpha and browning of white adipose tissues in adaptive thermogenesis. Gene Dev. 2012;26(3):271–81.PubMed CentralPubMedView ArticleGoogle Scholar
- Hotta Y, Nakamura H, Konishi M, Murata Y, Takagi H, Matsumura S, et al. Fibroblast growth factor 21 regulates lipolysis in white adipose tissue but is not required for ketogenesis and triglyceride clearance in liver. Endocrinology. 2009;150(10):4625–33.PubMedView ArticleGoogle Scholar
- Kharitonenkov A, Shiyanova TL, Koester A, Ford AM, Micanovic R, Galbreath EJ, et al. FGF-21 as a novel metabolic regulator. J Clin Invest. 2005;115(6):1627–35.PubMed CentralPubMedView ArticleGoogle Scholar
- Kharitonenkov A, Wroblewski VJ, Koester A, Chen YF, Clutinger CK, Tigno XT, et al. The metabolic state of diabetic monkeys is regulated by fibroblast growth factor-21. Endocrinology. 2007;148(2):774–81.PubMedView ArticleGoogle Scholar
- Lin Z, Tian H, Lam KSL, Lin S, Hoo RCL, Konishi M, et al. Adiponectin mediates the metabolic effects of FGF21 on glucose homeostasis and insulin sensitivity in mice. Cell Metab. 2013;17(5):779–89.PubMedView ArticleGoogle Scholar
- Sarruf DA, Thaler JP, Morton GJ, German J, Fischer JD, Ogimoto K, et al. Fibroblast growth factor 21 action in the brain increases energy expenditure and insulin sensitivity in obese rats. Diabetes. 2010;59(7):1817–24.PubMed CentralPubMedView ArticleGoogle Scholar
- Zhao Y, Dunbar JD, Kharitonenkov A. FGF21 as a therapeutic reagent. Adv Exp Med Biol. 2012;728:214–28.PubMedView ArticleGoogle Scholar
- Xu J, Stanislaus S, Chinookoswong N, Lau YY, Hager T, Patel J, et al. Acute glucose-lowering and insulin-sensitizing action of FGF21 in insulin-resistant mouse models-association with liver and adipose tissue effects. Am J Physiol Endocrinol Metab. 2009;297(5):E1105–14.PubMedView ArticleGoogle Scholar
- Berglund ED, Li CY, Bina HA, Lynes SE, Michael MD, Shanafelt AB, et al. Fibroblast Growth Factor 21 Controls Glycemia via Regulation of Hepatic Glucose Flux and Insulin Sensitivity. Endocrinology. 2009;150(9):4084–93.PubMed CentralPubMedView ArticleGoogle Scholar
- Shen Y, Ma X, Zhou J, Pan X, Hao Y, Zhou M, et al. Additive relationship between serum fibroblast growth factor 21 level and coronary artery disease. Cardiovasc Diabetol. 2013;12:124.PubMed CentralPubMedView ArticleGoogle Scholar
- Ong KL, Januszewski AS, O’Connell R, Jenkins AJ, Xu A, Sullivan DR, et al. The relationship of fibroblast growth factor 21 with cardiovascular outcome events in the Fenofibrate Intervention and Event Lowering in Diabetes study. Diabetologia. 2015;58(3):464–73.PubMedView ArticleGoogle Scholar
- Tacer KF, Bookout AL, Ding XS, Kurosu H, John GB, Wang L, et al. Research Resource: Comprehensive Expression Atlas of the Fibroblast Growth Factor System in Adult Mouse. Mol Endocrinol. 2010;24(10):2050–64.View ArticleGoogle Scholar
- Potthoff MJ, Inagaki T, Satapati S, Ding X, He T, Goetz R, et al. FGF21 induces PGC-1alpha and regulates carbohydrate and fatty acid metabolism during the adaptive starvation response. Proc Natl Acad Sci U S A. 2009;106(26):10853–8.PubMed CentralPubMedView ArticleGoogle Scholar
- Miao X, Wang YG, Sun J, Sun WX, Tan Y, Cai L, et al. Zinc protects against diabetes-induced pathogenic changes in the aorta: roles of metallothionein and nuclear factor (erythroid-derived 2)-like 2. Cardiovasc Diabetol. 2013;12:54.PubMed CentralPubMedView ArticleGoogle Scholar
- Miao X, Cui WP, Sun WX, Xin Y, Wang B, Tan Y, et al. Therapeutic Effect of MG132 on the Aortic Oxidative Damage and Inflammatory Response in OVE26 Type 1 Diabetic Mice. Oxid Med Cell Longev. 2013;2013:879516.PubMed CentralPubMedView ArticleGoogle Scholar
- Bai Y, Tan Y, Wang B, Miao X, Chen Q, Zheng Y, et al. Deletion of angiotensin II type 1 receptor gene or scavenge of superoxide prevents chronic alcohol-induced aortic damage and remodelling. J Cell Mol Med. 2012;16(10):2530–8.PubMed CentralPubMedView ArticleGoogle Scholar
- Jiang X, Zhang C, Xin Y, Huang Z, Tan Y, Huang Y, et al. Protective effect of FGF21 on type 1 diabetes-induced testicular apoptotic cell death probably via both mitochondrial–and endoplasmic reticulum stress-dependent pathways in the mouse model. Toxicol Lett. 2013;219(1):65–76.PubMedView ArticleGoogle Scholar
- Ignarro LJ, Buga GM, Wood KS, Byrns RE, Chaudhuri G. Endothelium-derived relaxing factor produced and released from artery and vein is nitric-oxide. Proc Natl Acad Sci U S A. 1987;84(24):9265–9.PubMed CentralPubMedView ArticleGoogle Scholar
- Huang PL. eNOS, metabolic syndrome and cardiovascular disease. Trends Endocrinol Metab. 2009;20(6):295–302.PubMed CentralPubMedView ArticleGoogle Scholar
- Kukreja RC, Xi L. eNOS phosphorylation: a pivotal molecular switch in vasodilation and cardioprotection? J Mol Cell Cardiol. 2007;42(2):280–2.PubMed CentralPubMedView ArticleGoogle Scholar
- Yan X, Chen J, Zhang C, Zhou S, Zhang Z, Chen J, Feng W, Li X, Tan Y: FGF21 deletion exacerbates diabetic cardiomyopathy by aggravating cardiac lipid accumulation. J Cell Mol Med 2015: 12530.Google Scholar
- Dimmeler S, Zeiher AM. Vascular repair by circulating endothelial progenitor cells: the missing link in atherosclerosis? J Mol Med (Berl). 2004;82(10):671–7.View ArticleGoogle Scholar
- Lu Y, Liu JH, Zhang LK, Du J, Zeng XJ, Hao G, et al. Fibroblast growth factor 21 as a possible endogenous factor inhibits apoptosis in cardiac endothelial cells. Chinese Med J-Peking. 2010;123(23):3417–21.Google Scholar
- Wang X-M, Song S-S, Xiao H, Gao P, Li X-J, Si L-Y. 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(3):658–71.PubMedView ArticleGoogle Scholar
- Zhu W, Wang C, Liu L, Li Y, Li X, Cai J, et al. Effects of fibroblast growth factor 21 on cell damage in vitro and atherosclerosis in vivo. Can J Physiol Pharmacol. 2014;92(11):927–35.PubMedView ArticleGoogle Scholar
- Lin Z, Pan X, Wu F, Ye D, Zhang Y, Wang Y, et al. 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(21):1861–71.PubMedView ArticleGoogle Scholar
- Insull Jr W. The pathology of atherosclerosis: plaque development and plaque responses to medical treatment. Am J Med. 2009;122(1 Suppl):S3–14.PubMedView ArticleGoogle Scholar
- Zhu S, Ma L, Wu Y, Ye X, Zhang T, Zhang Q, et al. FGF21 treatment ameliorates alcoholic fatty liver through activation of AMPK-SIRT1 pathway. Acta Biochim Biophys Sin (Shanghai). 2014;46(12):1041–8.View ArticleGoogle Scholar
- Ye D, Wang Y, Li H, Jia W, Man K, Lo CM, et al. Fibroblast growth factor 21 protects against acetaminophen-induced hepatotoxicity by potentiating peroxisome proliferator-activated receptor coactivator protein-1alpha-mediated antioxidant capacity in mice. Hepatology. 2014;60(3):977–89.PubMedView ArticleGoogle Scholar
- Kim HW, Lee JE, Cha JJ, Hyun YY, Kim JE, Lee MH, et al. Fibroblast growth factor 21 improves insulin resistance and ameliorates renal injury in db/db mice. Endocrinology. 2013;154(9):3366–76.PubMedView ArticleGoogle Scholar
- Zhang C, Shao ML, Yang H, Chen LM, Yu LC, Cong WT, et al. Attenuation of Hyperlipidemia–and Diabetes-Induced Early-Stage Apoptosis and Late-Stage Renal Dysfunction via Administration of Fibroblast Growth Factor-21 Is Associated with Suppression of Renal Inflammation. Plos One. 2013;8(12):e82275.PubMed CentralPubMedView ArticleGoogle Scholar
- Huang PL, Huang Z, Mashimo H, Bloch KD, Moskowitz MA, Bevan JA, et al. Hypertension in mice lacking the gene for endothelial nitric oxide synthase. Nature. 1995;377(6546):239–42.PubMedView ArticleGoogle Scholar
- Moroi M, Zhang L, Yasuda T, Virmani R, Gold HK, Fishman MC, et al. Interaction of genetic deficiency of endothelial nitric oxide, gender, and pregnancy in vascular response to injury in mice. J Clin Invest. 1998;101(6):1225–32.PubMed CentralPubMedView ArticleGoogle Scholar
- Lefer DJ, Jones SP, Girod WG, Baines A, Grisham MB, Cockrell AS, et al. Leukocyte-endothelial cell interactions in nitric oxide synthase-deficient mice. Am J Physiol. 1999;276(6 Pt 2):H1943–50.PubMedGoogle Scholar
- Freedman JE, Sauter R, Battinelli EM, Ault K, Knowles C, Huang PL, et al. Deficient platelet-derived nitric oxide and enhanced hemostasis in mice lacking the NOSIII gene. Circ Res. 1999;84(12):1416–21.PubMedView ArticleGoogle Scholar
- Kuhlencordt PJ, Gyurko R, Han F, Scherrer-Crosbie M, Aretz TH, Hajjar R, et al. Accelerated atherosclerosis, aortic aneurysm formation, and ischemic heart disease in apolipoprotein E/endothelial nitric oxide synthase double-knockout mice. Circulation. 2001;104(4):448–54.PubMedView ArticleGoogle Scholar
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