Heme oxygenase‑1 exerts pro‑apoptotic effects on hepatic stellate cells in vitro through regulation of nuclear factor‑κB

Yang,Hui; Chen,Bangtao; Zhao,Zhongfu; Zhang,Li; Zhang,Yun; Chen,Jie; Zhang,Xiaoqian; Zhang,Xiaohua; Zhao,Longfeng

doi:10.3892/etm.2018.6185

Heme oxygenase‑1 exerts pro‑apoptotic effects on hepatic stellate cells in vitro through regulation of nuclear factor‑κB

Authors:
- Hui Yang
- Bangtao Chen
- Zhongfu Zhao
- Li Zhang
- Yun Zhang
- Jie Chen
- Xiaoqian Zhang
- Xiaohua Zhang
- Longfeng Zhao
View Affiliations

Affiliations: Department of Infectious Diseases, The First Hospital of Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China, Department of Microbiology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, P.R. China, Institute of Hepatopathy, Changzhi Medical College, Changzhi, Shanxi 046011, P.R. China
Published online on: May 18, 2018 https://doi.org/10.3892/etm.2018.6185
Pages: 291-299
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Heme oxygenase-1 (HO-1) is an antioxidant and cytoprotective protein, which has been proven to alleviate the proliferation of hepatic stellate cells (HSCs) and the development of liver fibrosis. However, the role of HO‑1 in HSC apoptosis remains unclear. The aim of the present study was to investigate the effect of HO‑1 on HSC apoptosis and its possible underlying mechanisms. HSCs‑T6 were incubated with different concentrations of hemin (HO‑1 chemical inducer) and Znpp‑IX (HO‑1 chemical inhibitor) for 12, 24 and 48 h. Cell viability was determined using an MTT assay. HSCs were classified into 4 groups as follows: Control, hemin, Znpp‑IX and hemin+Znpp‑IX co‑treatment groups. Apoptosis was quantitatively measured by Annexin V/propidium iodide double staining and a terminal deoxynucleotidyl transferase dUTP nick‑end labeling assay. The mRNA and protein expression of HO‑1, α‑smooth muscle actin, B‑cell lymphoma (Bcl)‑2, caspase‑3 and nuclear factor (NF)‑κB p65 were measured using quantitative polymerase chain reaction and western blotting. The levels of tumor growth factor (TGF)‑β and interleukin (IL)‑6 in HSC supernatants were examined by ELISA. The results demonstrated that HO‑1 exerted antiproliferative effects on HSCs in a time‑ and concentration‑dependent manner. Increasing HO‑1 expression induced HSC apoptosis in vitro as demonstrated by a significant decrease in Bcl‑2 and an increase in caspase‑3 expression. Additionally, the expression of NF‑κB p65 and its downstream inflammatory factors TGF‑β and IL‑6 in the HO‑1 overexpression group was significantly decreased compared with the control group. Therefore, the present study provided evidence that HO‑1 serves an anti‑fibrosis role in the liver by enhancing HSC apoptosis, which was partially associated with the regulation of NF‑κB and its downstream effectors.

View Figures

View References

1	She H, Xiong S, Hazra S and Tsukamoto H: Adipogenic transcriptional regulation of hepatic stellate cells. J Biol Chem. 280:4959–4967. 2005. View Article : Google Scholar : PubMed/NCBI
2	Lee SH, Seo GS, Park YN and Sohn DH: Nephroblastoma overexpressed gene (NOV) expression in rat hepatic stellate cells. Biochem Pharmacol. 68:1391–1400. 2004. View Article : Google Scholar : PubMed/NCBI
3	Melhem A, Muhanna N, Bishara A, Alvarez CE, Ilan Y, Bishara T, Horani A, Nassar M, Friedman SL and Safadi R: Anti-fibrotic activity of NK cells in experimental liver injury through killing of activated HSC. J Hepatol. 45:60–71. 2006. View Article : Google Scholar : PubMed/NCBI
4	Burt AD: C. L. Oakley Lecture (1993). Cellular and molecular aspects of hepatic fibrosis. J Pathol. 170:105–114. 1993. View Article : Google Scholar : PubMed/NCBI
5	Di Sario A, Bendia E, Baroni Svegliati G, Ridolfi F, Casini A, Ceni E, Saccomanno S, Marzioni M, Trozzi L, Sterpetti P, et al: Effect of pirfenidone on rat hepatic stellate cell proliferation and collagen production. J Hepatol. 37:584–591. 2002. View Article : Google Scholar : PubMed/NCBI
6	Araujo JA, Zhang M and Yin F: Heme oxygenase-1, oxidation, inflammation, and atherosclerosis. Front Pharmacol. 3:1192012. View Article : Google Scholar : PubMed/NCBI
7	Gozzelino R, Jeney V and Soares MP: Mechanisms of cell protection by heme oxygenase-1. Annu Rev Pharmacol Toxicol. 50:323–354. 2010. View Article : Google Scholar : PubMed/NCBI
8	Kim KM, Im AR, Lee S and Chae S: Dual protective effects of flavonoids from petasites japonicus against UVB-induced apoptosis mediated via HSF-1 activated heat shock proteins and Nrf2-activated heme oxygenase-1 pathways. Biol Pharm Bull. 40:765–773. 2017. View Article : Google Scholar : PubMed/NCBI
9	Lijie Z, Ranran F, Xiuying L, Yutang H, Bo W and Tao M: Soyasaponin Bb protects rat hepatocytes from alcohol-induced oxidative stress by inducing heme oxygenase-1. Pharmacogn Mag. 12:302–306. 2016. View Article : Google Scholar : PubMed/NCBI
10	Park SW, Kang JW and Lee SM: The role of heme oxygenase-1 in drug metabolizing dysfunction in the alcoholic fatty liver exposed to ischemic injury. Toxicol Appl Pharmacol. 292:30–39. 2016. View Article : Google Scholar : PubMed/NCBI
11	Wu B, Song HL, Yang Y, Yin ML, Zhang BY, Cao Y, Dong C and Shen ZY: Improvement of liver transplantation outcome by heme oxygenase-1-transduced bone marrow mesenchymal stem cells in rats. Stem Cells Int. 2016:92350732016. View Article : Google Scholar : PubMed/NCBI
12	Yang H, Zhao LF, Zhao ZF, Wang Y, Zhao JJ and Zhang L: Heme oxygenase-1 prevents liver fibrosis in rats by regulating the expression of PPARγ and NF-κB. World J Gastroenterol. 18:1680–1688. 2012. View Article : Google Scholar : PubMed/NCBI
13	Supinski GS and Callahan LA: Hemin prevents cardiac and diaphragm mitochondrial dysfunction in sepsis. Free Radic Biol Med. 40:127–137. 2006. View Article : Google Scholar : PubMed/NCBI
14	Liu H, Song D and Lee SS: Role of heme oxygenase-carbon monoxide pathway in pathogenesis of cirrhotic cardiomyopathy in the rat. Am J Physiol Gastrointest Liver Physiol. 280:G68–G74. 2001. View Article : Google Scholar : PubMed/NCBI
15	Martasek P, Schwartzman ML, Goodman AI, Solangi KB, Levere RD and Abraham NG: Hemin and L-arginine regulation of blood pressure in spontaneously hypertensive rats. J Am Soc Nephrol. 2:1078–1084. 1991.PubMed/NCBI
16	Hualin C, Wenli X, Dapeng L, Xijing L, Xiuhua P and Qingfeng P: The anti-inflammatory mechanism of heme oxygenase-1 induced by hemin in primary rat alveolar macrophages. Inflammation. 35:1087–1093. 2012. View Article : Google Scholar : PubMed/NCBI
17	Choi KM, Gibbons SJ, Nguyen TV, Stoltz GJ, Lurken MS, Ordog T, Szurszewski JH and Farrugia G: Heme oxygenase-1 protects interstitial cells of Cajal from oxidative stress and reverses diabetic gastroparesis. Gastroenterology. 135:2055–2064. 2008. View Article : Google Scholar : PubMed/NCBI
18	Li X, Wang Y, Wang H, Huang C, Huang Y and Li J: Endoplasmic reticulum stress is the crossroads of autophagy, inflammation, and apoptosis signaling pathways and participates in liver fibrosis. Inflamm Res. 64:1–7. 2015. View Article : Google Scholar : PubMed/NCBI
19	Gressner AM: The up-and-down of hepatic stellate cells in tissue injury: Apoptosis restores cellular homeostasis. Gastroenterology. 120:1285–1288. 2001. View Article : Google Scholar : PubMed/NCBI
20	Inguaggiato P, Gonzalez-Michaca L, Croatt AJ, Haggard JJ, Alam J and Nath KA: Cellular overexpression of heme oxygenase-1 up-regulates p21 and confers resistance to apoptosis. Kidney Int. 60:2181–2191. 2001. View Article : Google Scholar : PubMed/NCBI
21	Soares MP, Usheva A, Brouard S, Berberat PO, Gunther L, Tobiasch E and Bach FH: Modulation of endothelial cell apoptosis by heme oxygenase-1-derived carbon monoxide. Anti oxid Redox Signal. 4:321–329. 2002. View Article : Google Scholar
22	Nan Y, Wang R, Zhao S, Han F, Wu WJ, Kong L, Fu N, Kong L and Yu J: Heme oxygenase-1 prevents non-alcoholic steatohepatitis through suppressing hepatocyte apoptosis in mice. Lipids Health Dis. 9:1242010. View Article : Google Scholar : PubMed/NCBI
23	Aung KH, Win-Shwe TT, Kanaya M, Takano H and Tsukahara S: Involvement of hemeoxygenase-1 in di(2-ethylhexyl) phthalate (DEHP)-induced apoptosis of Neuro-2a cells. J Toxicol Sci. 39:217–229. 2014. View Article : Google Scholar : PubMed/NCBI
24	Liu XM, Chapman GB, Wang H and Durante W: Adenovirus-mediated heme oxygenase-1 gene expression stimulates apoptosis in vascular smooth muscle cells. Circulation. 105:79–84. 2002. View Article : Google Scholar : PubMed/NCBI
25	Lin CW, Shen SC, Hou WC, Yang LY and Chen YC: Heme oxygenase-1 inhibits breast cancer invasion via suppressing the expression of matrix metalloproteinase-9. Mol Cancer Ther. 7:1195–1206. 2008. View Article : Google Scholar : PubMed/NCBI
26	Lee WY, Chen YC, Shih CM, Lin CM, Cheng CH, Chen KC and Lin CW: The induction of heme oxygenase-1 suppresses heat shock protein 90 and the proliferation of human breast cancer cells through its byproduct carbon monoxide. Toxicol Appl Pharmacol. 274:55–62. 2014. View Article : Google Scholar : PubMed/NCBI
27	Lee HN, Jin HO, Park JA, Kim JH, Kim JY, Kim B, Kim W, Hong SE, Lee YH, Chang YH, et al: Heme oxygenase-1 determines the differential response of breast cancer and normal cells to piperlongumine. Mol Cells. 38:327–335. 2015. View Article : Google Scholar : PubMed/NCBI
28	Mòdol T, Brice N, de Galarreta Ruiz M, Garzón García A, Iraburu MJ, Martínez-Irujo JJ and López-Zabalza MJ: Fibronectin peptides as potential regulators of hepatic fibrosis through apoptosis of hepatic stellate cells. J Cell Physiol. 230:546–553. 2015. View Article : Google Scholar : PubMed/NCBI
29	Elsharkawy AM and Mann DA: Nuclear factor-kappaB and the hepatic inflammation-fibrosis-cancer axis. Hepatology. 46:590–597. 2007. View Article : Google Scholar : PubMed/NCBI
30	Uwagawa T and Yanaga K: Effect of NF-κB inhibition on chemoresistance in biliary-pancreatic cancer. Surg Today. 45:1481–1488. 2015. View Article : Google Scholar : PubMed/NCBI
31	Kucharczak J, Simmons MJ, Fan Y and Gélinas C: To be, or not to be: NF-kappaB is the answer-role of Rel/NF-kappaB in the regulation of apoptosis. Oncogene. 22:8961–8982. 2003. View Article : Google Scholar : PubMed/NCBI
32	Son G, Iimuro Y, Seki E, Hirano T, Kaneda Y and Fujimoto J: Selective inactivation of NF-kappaB in the liver using NF-kappaB decoy suppresses CCl4-induced liver injury and fibrosis. Am J Physiol Gastrointest Liver Physiol. 293:G631–G639. 2007. View Article : Google Scholar : PubMed/NCBI
33	Lin CC, Chiang LL, Lin CH, Shih CH, Liao YT, Hsu MJ and Chen BC: Transforming growth factor-beta1 stimulates heme oxygenase-1 expression via the PI3K/Akt and NF-kappaB pathways in human lung epithelial cells. Eur J Pharmacol. 560:101–109. 2007. View Article : Google Scholar : PubMed/NCBI
34	Zhang L, Zhang Z, Liu B, Jin Y, Tian Y, Xin Y and Duan Z: The protective effect of heme oxygenase-1 against intestinal barrier dysfunction in cholestatic liver injury is associated with NF-κB inhibition. Mol Med. 23:2017.(Epub ahead of print). View Article : Google Scholar
35	Yeh CH, Chen TP, Wang YC, Lin YM and Lin PJ: HO-1 activation can attenuate cardiomyocytic apoptosis via inhibition of NF-kappaB and AP-1 translocation following cardiac global ischemia and reperfusion. J Surg Res. 155:147–156. 2009. View Article : Google Scholar : PubMed/NCBI
36	So H, Kim H, Kim Y, Kim E, Pae HO, Chung HT, Kim HJ, Kwon KB, Lee KM, Lee HY, et al: Evidence that cisplatin-induced auditory damage is attenuated by downregulation of pro-inflammatory cytokines via Nrf2/HO-1. J Assoc Res Otolaryngol. 9:290–306. 2008. View Article : Google Scholar : PubMed/NCBI