FGF21 attenuates hypoxia‑induced dysfunction and apoptosis in HPAECs through alleviating endoplasmic reticulum stress

Chen,Ali; Liu,Jingjing; Zhu,Jianfeng; Wang,Xuetao; Xu,Zhaona; Cui,Zhimin; Yao,Dan; Huang,Zhifeng; Xu,Min; Chen,Mayun; Wu,Peiliang; Li,Manxiang; Wang,Liangxing; Huang,Xiaoying

doi:10.3892/ijmm.2018.3705

FGF21 attenuates hypoxia‑induced dysfunction and apoptosis in HPAECs through alleviating endoplasmic reticulum stress

Authors:
- Ali Chen
- Jingjing Liu
- Jianfeng Zhu
- Xuetao Wang
- Zhaona Xu
- Zhimin Cui
- Dan Yao
- Zhifeng Huang
- Min Xu
- Mayun Chen
- Peiliang Wu
- Manxiang Li
- Liangxing Wang
- Xiaoying Huang
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Affiliations: Division of Pulmonary Medicine, The First Affiliated Hospital of Wenzhou Medical University, Key Laboratory of Heart and Lung, Wenzhou, Zhejiang 325000, P.R. China, Key Laboratory of Biotechnology and Pharmaceutical Engineering of Zhejiang Province, Wenzhou Medical University, Wenzhou, Zhejiang 325000, P.R. China, Department of Respiratory Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shanxi 710061, P.R. China
Published online on: May 24, 2018 https://doi.org/10.3892/ijmm.2018.3705
Pages: 1684-1694

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Abstract

Vascular endothelial apoptosis and dysfunction have a crucial role in triggering pathological vascular remodeling of hypoxia‑induced pulmonary arterial hypertension (PAH). Fibroblast growth factor (FGF)21, an endocrine regulator, has recently been reported to protect cardiac endothelial cells from damage and suppress inflammatory responses. In addition, FGF21 is reported to be involved in endoplasmic reticulum stress (ERS). Previous studies have suggested that ERS participates in the development of PAH, and attenuation of ERS could be an effective therapeutic strategy for the protection of pulmonary arteries. However, whether FGF21 has a protective function via suppression of ERS in pulmonary arterial endothelial cells in hypoxia remains unclear. The present study aimed to explore whether FGF21 could reduce the hypoxia‑induced apoptosis of human pulmonary arterial endothelial cells (HPAECs) and prevent endothelial dysfunction via the inhibition of ERS. HPAECs were divided into six groups: Normoxia, hypoxia, hypoxia plus FGF21, hypoxia plus salubrinal (an ERS inhibitor), hypoxia plus tunicamycin (an ERS agonist), and hypoxia plus tunicamycin plus FGF21. The endoplasmic reticulum ultrastructure in HPAECs was assessed by transmission electron microscopy, and proliferation and apoptosis were examined by cell counting kit‑8 and terminal deoxyribonucleotide transferase‑mediated dUTP nick end‑labelling assays, respectively. The expression levels of ERS‑related proteins, including binding immunoglobulin protein (BiP), protein kinase R‑like endoplasmic reticulum kinase (PERK), phosphorylated (p‑) PERK, transcription factor C/EBP homologous protein (CHOP), B‑cell lymphoma-2 (Bcl‑2) and caspase‑4 were detected by western blotting. Transwell migration chamber assays were performed, and the concentration of nitric oxide (NO)/endothelin‑1 (ET‑1) in the culture medium was determined to examine endothelial function. The results revealed that hypoxia increased the % of apoptotic cells and diminished the viability of HPAECs, accompanied by an upregulation of ERS‑dependent apoptosis by increasing the expression of the proapoptotic caspase‑4 and decreasing the antiapoptotic Bcl‑2. Additionally, hypoxia upregulated the expression of representative proteins in the PERK branch of ERS, including BiP, p‑PERK and CHOP, while it downregulated the expression of PERK. Furthermore, the secretion of NO/ET‑1 and the migration rate of HPAECs were downregulated under conditions of hypoxia. FGF21 significantly attenuated the hypoxia‑induced apoptosis and dysfunction of HPAECs through alleviating the aforementioned changes in ERS‑dependent signaling pathways. In conclusion, ERS may be a crucial mechanism in the hypoxia‑induced apoptosis and endothelial dysfunction of HPAECs. FGF21 may attenuate the hypoxia‑induced apoptosis and dysfunction of HPAECs through alleviating ERS, via the PERK/CHOP signaling pathway and inhibition of caspase‑4 expression.

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1	Sutendra G and Michelakis ED: The metabolic basis of pulmonary arterial hypertension. Cell Metab. 19:558–573. 2014. View Article : Google Scholar : PubMed/NCBI
2	Huang X, Zou L, Yu X, Chen M, Guo R, Cai H, Yao D, Xu X, Chen Y, Ding C, et al: Salidroside attenuates chronic hypoxia-induced pulmonary hypertension via adenosine A2a receptor related mitochondria-dependent apoptosis pathway. J Mol Cell Cardiol. 82:153–166. 2015. View Article : Google Scholar : PubMed/NCBI
3	Voelkel NF and Cool C: Pathology of pulmonary hypertension. Cardiol Clin. 22:343–351. 2004. View Article : Google Scholar : PubMed/NCBI
4	Eddahibi S, Guignabert C, Barlier-Mur AM, Dewachter L, Fadel E, Dartevelle P, Humbert M, Simonneau G, Hanoun N, Saurini F, et al: Cross talk between endothelial and smooth muscle cells in pulmonary hypertension: Critical role for serotonin-induced smooth muscle hyperplasia. Circulation. 113:1857–1864. 2006. View Article : Google Scholar : PubMed/NCBI
5	Humbert M, Montani D, Perros F, Dorfmüller P, Adnot S and Eddahibi S: Endothelial cell dysfunction and cross talk between endothelium and smooth muscle cells in pulmonary arterial hypertension. Vascul Pharmacol. 49:113–118. 2008. View Article : Google Scholar : PubMed/NCBI
6	Sahara M, Sata M, Morita T, Hirata Y and Nagai R: Nicorandil attenuates monocrotaline-induced vascular endothelial damage and pulmonary arterial hypertension. PLoS One. 7:e333672012. View Article : Google Scholar : PubMed/NCBI
7	Farkas L, Farkas D, Ask K, Möller A, Gauldie J, Margetts P, Inman M and Kolb M: VEGF ameliorates pulmonary hypertension through inhibition of endothelial apoptosis in experimental lung fibrosis in rats. J Clin Invest. 119:1298–1311. 2009. View Article : Google Scholar : PubMed/NCBI
8	Wan XS, Lu XH, Xiao YC, Lin Y, Zhu H, Ding T, Yang Y, Huang Y, Zhang Y, Liu YL, et al: ATF4- and CHOP-dependent induction of FGF21 through endoplasmic reticulum stress. Biomed Res Int. 2014:8078742014. View Article : Google Scholar : PubMed/NCBI
9	Walter P and Ron D: The unfolded protein response: From stress pathway to homeostatic regulation. Science. 334:1081–1086. 2011. View Article : Google Scholar : PubMed/NCBI
10	Muñoz JP and Zorzano A: Endoplasmic reticulum stress enters a Nogo Zone. Sci Transl Med. 3:88ps262011. View Article : Google Scholar : PubMed/NCBI
11	Koyama M, Furuhashi M, Ishimura S, Mita T, Fuseya T, Okazaki Y, Yoshida H, Tsuchihashi K and Miura T: Reduction of endoplasmic reticulum stress by 4-phenylbutyric acid prevents the development of hypoxia-induced pulmonary arterial hypertension. Am J Physiol Heart Circ Physiol. 306:H1314–H1323. 2014. View Article : Google Scholar : PubMed/NCBI
12	Fan XF, Li WJ, Chen ZQ, Wang XR, Kong XX, Mao SZ, Hu LG and Gong YS: Changes of endoplasmic reticulum stress-induced apoptosis in pulmonary tissue of rats with hypoxic pulmonary hypertension. Zhongguo Ying Yong Sheng Li Xue Za Zhi. 27:270–274. 2011.In Chinese. PubMed/NCBI
13	Ost M, Coleman V, Kasch J and Klaus S: Regulation of myokine expression: Role of exercise and cellular stress. Free Radic Biol Med. 98:78–89. 2016. View Article : Google Scholar : PubMed/NCBI
14	Inagaki T: Research perspectives on the regulation and physiological functions of FGF21 and its association with NAFLD. Front Endocrinol (Lausanne). 6:1472015.
15	Kim SH, Kim KH, Kim HK, Kim MJ, Back SH, Konishi M, Itoh N and Lee MS: Fibroblast growth factor 21 participates in adaptation to endoplasmic reticulum stress and attenuates obesity-induced hepatic metabolic stress. Diabetologia. 58:809–818. 2015. View Article : Google Scholar
16	Shimizu M, Morimoto H, Maruyama R, Inoue J and Sato R: Selective regulation of FGF19 and FGF21 expression by cellular and nutritional stress. J Nutr Sci Vitaminol (Tokyo). 61:154–160. 2015. View Article : Google Scholar
17	Guo Q, Xu L, Liu J, Li H, Sun H, Wu S and Zhou B: Fibroblast growth factor 21 reverses suppression of adiponectin expression via inhibiting endoplasmic reticulum stress in adipose tissue of obese mice. Exp Biol Med (Maywood). 242:441–447. 2017. View Article : Google Scholar
18	Lü Y, Liu JH, Zhang LK, DU J, Zeng XJ, Hao G, Huang J, Zhao DH, Wang GZ and Zhang YC: Fibroblast growth factor 21 as a possible endogenous factor inhibits apoptosis in cardiac endothelial cells. Chin Med J (Engl). 123:3417–3421. 2010.
19	Li J, Zhou J, Zhang D, Song Y, She J and Bai C: Bone marrow-derived mesenchymal stem cells enhance autophagy via PI3K/AKT signalling to reduce the severity of ischaemia/reper-fusion-induced lung injury. J Cell Mol Med. 19:2341–2351. 2015. View Article : Google Scholar : PubMed/NCBI
20	Gong T, Wang Q, Lin Z, Chen ML and Sun GZ: Endoplasmic reticulum (ER) stress inhibitor salubrinal protects against ceramide-induced SH-SY5Y cell death. Biochem Biophys Res Commun. 427:461–465. 2012. View Article : Google Scholar : PubMed/NCBI
21	Guoliang H, Kunlun H, Li F, Xin L and Ruijun L: Salubrinal protects endoplasmic reticulum of cardiac muscle cells against stress-associated apoptosis. Jun Yi Jin Xiu Xue Yuan Xue Bao. 31:483–485. 2010.In Chinese.
22	Wang H, Zuo X, Wang Q, Yu Y, Xie L, Wang H, Wu H and Xie W: Nicorandil inhibits hypoxia-induced apoptosis in human pulmonary artery endothelial cells through activation of mito-KATP and regulation of eNOS and the NF-κB pathway. Int J Mol Med. 32:187–194. 2013. View Article : Google Scholar : PubMed/NCBI
23	Morecroft I, White K, Caruso P, Nilsen M, Loughlin L, Alba R, Reynolds PN, Danilov SM, Baker AH and Maclean MR: Gene therapy by targeted adenovirus-mediated knockdown of pulmonary endothelial Tph1 attenuates hypoxia-induced pulmonary hypertension. Mol Ther. 20:1516–1528. 2012. View Article : Google Scholar : PubMed/NCBI
24	Paulin R and Michelakis ED: The metabolic theory of pulmonary arterial hypertension. Circ Res. 115:148–164. 2014. View Article : Google Scholar : PubMed/NCBI
25	Mao SZ, Fan XF, Xue F, Chen R, Chen XY, Yuan GS, Hu LG, Liu SF and Gong YS: Intermedin modulates hypoxic pulmonary vascular remodeling by inhibiting pulmonary artery smooth muscle cell proliferation. Pulm Pharmacol Ther. 27:1–9. 2014. View Article : Google Scholar
26	Wang S and Kaufman RJ: The impact of the unfolded protein response on human disease. J Cell Biol. 197:857–867. 2012. View Article : Google Scholar : PubMed/NCBI
27	Liu Z, Lv Y, Zhao N, Guan G and Wang J: Protein kinase R-like ER kinase and its role in endoplasmic reticulum stress-decided cell fate. Cell Death Dis. 6:e18222015. View Article : Google Scholar : PubMed/NCBI
28	Vannuvel K, Renard P, Raes M and Arnould T: Functional and morphological impact of ER stress on mitochondria. J Cell Physiol. 228:1802–1818. 2013. View Article : Google Scholar : PubMed/NCBI
29	Rutkowski DT and Kaufman RJ: A trip to the ER: Coping with stress. Trends Cell Biol. 14:20–28. 2004. View Article : Google Scholar : PubMed/NCBI
30	Szegezdi E, Fitzgerald U and Samali A: Caspase-12 and ER-stress-mediated apoptosis: The story so far. Ann NY Acad Sci. 1010:186–194. 2003. View Article : Google Scholar
31	Richardson CE, Kinkel S and Kim DH: Physiological IRE-1-XBP-1 and PEK-1 signaling in caenorhabditis elegans larval development and immunity. PLoS Genet. 7:e10023912011. View Article : Google Scholar : PubMed/NCBI
32	Senkal CE, Ponnusamy S, Manevich Y, Meyers-Needham M, Saddoughi SA, Mukhopadyay A, Dent P, Bielawski J and Ogretmen B: Alteration of ceramide synthase 6/C16-ceramide induces activating transcription factor 6-mediated endoplasmic reticulum (ER) stress and apoptosis via perturbation of cellular Ca² and ER/Golgi membrane network. J Biol Chem. 286:42446–42458. 2011. View Article : Google Scholar : PubMed/NCBI
33	Schröder M and Kaufman RJ: ER stress and the unfolded protein response. Mutat Res. 569:29–63. 2005. View Article : Google Scholar
34	Boyce M, Bryant KF, Jousse C, Long K, Harding HP, Scheuner D, Kaufman RJ, Ma D, Coen DM, Ron D and Yuan J: A Selective Inhibitor of eIF2alpha dephosphorylation protects cells from ER stress. Science. 307:935–939. 2005. View Article : Google Scholar : PubMed/NCBI
35	Wu Y, Adi D, Long M, Wang J, Liu F, Gai MT, Aierken A, Li MY, Li Q, Wu LQ, et al: 4-Phenylbutyric acid induces protection against pulmonary arterial hypertension in rats. PLoS One. 11:e01575382016. View Article : Google Scholar : PubMed/NCBI
36	Ornitz DM and Itoh N: Fibroblast growth factors. Genome Biol. 2:REVIEWS3005. 2001. View Article : Google Scholar : PubMed/NCBI
37	Schaap FG, Kremer AE, Lamers WH, Jansen PL and Gaemers IC: Fibroblast growth factor 21 is induced by endoplasmic reticulum stress. Biochimie. 95:692–699. 2013. View Article : Google Scholar
38	Jiang S, Yan C, Fang QC, Shao ML, Zhang YL, Liu Y, Deng YP, Shan B, Liu JQ, Li HT, et al: Fibroblast growth factor 21 is regulated by the IRE1α-XBP1 branch of the unfolded protein response and counteracts endoplasmic reticulum stress-induced hepatic steatosis. J Biol Chem. 289:29751–29765. 2014. View Article : Google Scholar : PubMed/NCBI
39	Budhiraja R, Tuder RM and Hassoun PM: Endothelial dysfunction in pulmonary hypertension. Circulation. 109:159–165. 2004. View Article : Google Scholar : PubMed/NCBI
40	Marletta MA: Nitric oxide synthase structure and mechanism. J Biol Chem. 268:12231–12234. 1993.PubMed/NCBI
41	Ozaki M, Kawashima S, Yamashita T, Ohashi Y, Rikitake Y, Inoue N, Hirata KI, Hayashi Y, Itoh H and Yokoyama M: Reduced hypoxic pulmonary vascular remodeling by nitric oxide from the endothelium. Hypertension. 37:322–327. 2001. View Article : Google Scholar : PubMed/NCBI
42	Rubin LJ, Badesch DB, Barst RJ, Galie N, Black CM, Keogh A, Pulido T, Frost A, Roux S, Leconte I, et al: Bosentan therapy for pulmonary arterial hypertension. N Engl J Med. 346:896–903. 2002. View Article : Google Scholar : PubMed/NCBI