Transforming growth factor‑β1 reduces apoptosis via autophagy activation in hepatic stellate cells

Fu,Mei‑Ya; He,Ya‑Jun; Lv,Xia; Liu,Zhi‑He; Shen,Yan; Ye,Guo‑Rong; Deng,Yan‑Mei; Shu,Jian‑Chang

doi:10.3892/mmr.2014.2383

Transforming growth factor‑β1 reduces apoptosis via autophagy activation in hepatic stellate cells

Authors:
- Mei‑Ya Fu
- Ya‑Jun He
- Xia Lv
- Zhi‑He Liu
- Yan Shen
- Guo‑Rong Ye
- Yan‑Mei Deng
- Jian‑Chang Shu
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Affiliations: Department of Gastroenterology, The Fourth Affiliated Hospital of the Medical College of Jinan University, Guangzhou Red Cross Hospital, Guangzhou, Guangdong 510220, P.R. China
Published online on: July 15, 2014 https://doi.org/10.3892/mmr.2014.2383
Pages: 1282-1288
Copyright: © Fu et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Autophagy is a metabolic process that is important in fibrogenesis, in which cellular components are degraded by lysosomal machinery. Transforming growth factor β1 (TGF‑β1) is a potent fibrogenic cytokine involved in liver fibrosis; however, it remains elusive whether autophagy is regulated by TGF‑β1 in this process. In the present study, the function of TGF‑β1‑mediated autophagy in the proliferation and apoptosis of hepatic stellate cells (HSCs) was investigated. A rat HSC cell line (HSC‑T6) was incubated with or without TGF‑β1 followed by bafilomycin A1, and microtubule-associated proteins 1A/1B light chain 3 (LC3) small interfering (si)RNA was used to inhibit autophagy in order to assess the association between TGF‑β1 and autophagy. HSC‑T6 cell transient transfection was accomplished with a pLVX‑AcGFP‑N1‑rLC3B‑encoding plasmid. An MTS assay and flow cytometry were utilized to detect proliferation and apoptosis of HSC‑T6 cells. Quantitative polymerase chain reaction, immunoﬂuorescence and western blot analysis were used to detect the presence of activation markers. Proliferation was increased and apoptosis was reduced in HSC‑T6 cells treated with TGF‑β1 compared with cells subjected to serum deprivation. However, when HSC‑T6 cells were treated with bafilomycin A1 and LC3 siRNA, increased apoptosis and reduced proliferation were observed. In addition, protein and mRNA expression levels of the autophagy marker LC3 were significantly increased. GFP‑LC3 punctate markings were more prolific following TGF‑β1 treatment of HSC‑T6 cells, indicating that TGF‑β1 may rescue HSC‑T6 cells from serum deprivation and reduce apoptosis via autophagy induction. The present study elucidated the possible functions of TGF‑β1‑mediated autophagy in the pathological process of liver fibrosis.

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1	Ni HM, Williams JA, Yang H, Shi YH, Fan J and Ding WX: Targeting autophagy for the treatment of liver diseases. Pharmacol Res. 66:463–474. 2012. View Article : Google Scholar : PubMed/NCBI
2	Klionsky DJ: The autophagy connection. Dev Cell. 19:11–12. 2010. View Article : Google Scholar
3	González-Polo RA, Boya P, Pauleau AL, et al: The apoptosis/autophagy paradox: autophagic vacuolization before apoptotic death. J Cell Sci. 118:3091–3102. 2005.PubMed/NCBI
4	Komatsu M: Liver autophagy: physiology and pathology. J Bio Chem. 152:5–15. 2012.PubMed/NCBI
5	Sasaki M, Miyakoshi M, Sato Y and Nakanuma Y: Autophagy may precede cellular senescence of bile ductular cells in ductular reaction in primary biliary cirrhosis. Dig Dis Sci. 57:660–666. 2012. View Article : Google Scholar
6	Bhogal RH and Afford SC: Autophagy and the Liver. Autophagy - A Double-Edged Sword - Cell Survival or Death? Bailly Yannick: InTech; Rijeka, Croatia: pp. 165–185. 2013
7	Rautou PE, Mansouri A, Lebrec D, Durand F, Valla D and Moreau R: Autophagy in liver diseases. J Hepatol. 53:1123–1134. 2010. View Article : Google Scholar
8	Hilscher M, Hernandez-Gea V and Friedman SL: Autophagy and mesenchymal cell fibrogenesis. Biochim Biophys Acta. 1831.972–978. 2012.PubMed/NCBI
9	Hidvegi T, Ewing M, Hale P, et al: An autophagy-enhancing drug promotes degradation of mutant alpha1-antitrypsin Z and reduces hepatic fibrosis. Science. 329:229–232. 2010. View Article : Google Scholar : PubMed/NCBI
10	Kim SI, Na HJ, Ding Y, Wang Z, Lee SJ and Choi ME: Autophagy promotes intracellular degradation of type I collagen induced by transforming growth factor (TGF)-β1. J Biol Chem. 287:11677–11688. 2012.PubMed/NCBI
11	Hernández-Gea V, Ghiassi-Nejad Z, Rozenfeld R, et al: Autophagy releases lipid that promotes fibrogenesis by activated hepatic stellate cells in mice and in human tissues. Gastroenterology. 142:938–946. 2012.PubMed/NCBI
12	Thoen LF, Guimarães EL, Dollé L, et al: A role for autophagy during hepatic stellate cell activation. J Hepatol. 55:1353–1360. 2011. View Article : Google Scholar : PubMed/NCBI
13	Yin XM, Ding WX and Gao W: Autophagy in the liver. Hepatology. 47:1773–1785. 2008. View Article : Google Scholar : PubMed/NCBI
14	Rubinsztein DC, Gestwicki JE, Murphy LO and Klionsky DJ: Potential therapeutic applications of autophagy. Nat Rev Drug Discov. 6:304–312. 2007. View Article : Google Scholar : PubMed/NCBI
15	Deretic V and Levine B: Autophagy, immunity and microbial adaptations. Cell Host Microbe. 5:527–549. 2009. View Article : Google Scholar : PubMed/NCBI
16	Friedman SL: Hepatic fibrosis - overview. Toxicology. 254:120–129. 2008. View Article : Google Scholar
17	Brenner DA: Molecular pathogenesis of liver fibrosis. Trans Am Clin Climatol Assoc. 120:361–368. 2009.PubMed/NCBI
18	Iimuro Y and Brenner DA: Matrix metalloproteinase gene delivery for liver fibrosis. Pharm Res. 25:249–258. 2008. View Article : Google Scholar : PubMed/NCBI
19	Gajewska M, Gajkowska B and Motyl T: Apoptosis and autophagy induced by TGF-β1 in bovine mammary epithelial BME-UV1 cells. J Physiol Pharmacol. 56(Suppl 3): 143–157. 2005.
20	Ding Y, Kim JK, Kim SI, Na HJ, Jun SY, Lee SJ and Choi ME: TGF-{beta}1 protects against mesangial cell apoptosis via induction of autophagy. J Biol Chem. 285:37909–37919. 2010. View Article : Google Scholar : PubMed/NCBI
21	Klionsky DJ, Abdalla FC, Abeliovich H, et al: Guidelines for the use and interpretation of assays for monitoring autophagy. Autophagy. 8:445–544. 2012. View Article : Google Scholar
22	Leask A and Abraham DJ: TGF beta signaling and the fibrotic response. FASEB J. 18:816–827. 2004. View Article : Google Scholar : PubMed/NCBI
23	Nakerakanti S and Trojanowska M: The role of TGF-β receptors in fibrosis. Open Rheumatol J. 6:156–162. 2012.
24	Roberts AB: Molecular and cell biology of TGF-beta. Miner Electrolyte Metab. 24:111–119. 1998. View Article : Google Scholar
25	Roberts AB and Sporn MB: Physiological actions and clinical applications of transforming growth factor-beta (TGF-beta). Growth Factors. 8:1–9. 1993. View Article : Google Scholar : PubMed/NCBI
26	Brown TL, Patil S, Cianci CD, Morrow JS and Howe PH: Transforming growth factor-beta induces caspase 3-independent cleavage of alphaII-spectrin (alpha-fodrin) coincident with apoptosis. J Biol Chem. 274:23256–23262. 1999. View Article : Google Scholar : PubMed/NCBI
27	Dai C, Yang J and Liu Y: Transforming growth factor-beta1 potentiates renal tubular epithelial cell death by a mechanism independent of Smad signaling. J Biol Chem. 278:12537–12545. 2003. View Article : Google Scholar : PubMed/NCBI
28	Chin BY, Petrache I, Choi AM and Choi ME: Transforming growth factor beta1 rescues serum deprivation-induced apoptosis via the mitogen-activated protein kinase (MAPK) pathway in macrophages. J Biol Chem. 274:11362–11368. 1999. View Article : Google Scholar : PubMed/NCBI
29	Hara T, Kamura T and Nakayama K, Oshikawa K, Hatakeyama S and Nakayama K: Degradation of p27(Kip1) at the G(0)–G(1) transition mediated by a Skp2-independent ubiquitination pathway. J Biol Chem. 276:48937–48943. 2001.PubMed/NCBI
30	Chen HH, Zhao S and Song JG: TGF-beta1 suppresses apoptosis via differential regulation of MAP kinases and ceramide production. Cell Death Differ. 10:516–527. 2003. View Article : Google Scholar : PubMed/NCBI
31	Maiuri MC, Zalckvar E, Kimchi A and Kroemer G: Self-eating and self-killing: crosstalk between autophagy and apoptosis. Nat Rev Mol Cell Biol. 8:741–752. 2007. View Article : Google Scholar : PubMed/NCBI
32	Wang RC and Levine B: Autophagy in cellular growth control. FEBS Lett. 584:1417–1426. 2010. View Article : Google Scholar : PubMed/NCBI
33	Suzuki HI, Kiyono K and Miyazono K: Regulation of autophagy by transforming growth factor-β (TGF-β) signaling. Autophagy. 6:645–647. 2010.