Protection of cultured human hepatocytes from hydrogen peroxide‑induced apoptosis by relaxin‑3

Ma,Xiao; Han,Su; Zhang,Wei; Fan,Yu‑Jing; Liu,Ming‑Na; Liu,Ai‑Yun; Liu,Bing‑Rong

doi:10.3892/mmr.2014.2842

Protection of cultured human hepatocytes from hydrogen peroxide‑induced apoptosis by relaxin‑3

Authors:
- Xiao Ma
- Su Han
- Wei Zhang
- Yu‑Jing Fan
- Ming‑Na Liu
- Ai‑Yun Liu
- Bing‑Rong Liu
View Affiliations

Affiliations: Department of Gastroenterology and Hepatology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China, Department of Microbiology and Parasitology, The Heilongjiang Key Laboratory of Immunity and Infection, Pathogenic Biology, Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China, Center for Endemic Disease Control, Chinese Center for Disease Control and Prevention, Harbin Medical University, Harbin, Heilongjiang 150081, P.R. China
Published online on: November 3, 2014 https://doi.org/10.3892/mmr.2014.2842
Pages: 1228-1234

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

Previous studies have suggested that hepatocyte apoptosis may be a fundamental underlying mechanism of liver injury and diseases, such as liver fibrosis. Relaxin‑3 has been reported to have anti‑fibrotic actions in the heart and to attenuate isoproterenol‑induced myocardial injury; however, the beneficial role of relaxin‑3 on hepatocyte apoptosis remains to be elucidated. The aim of the present study was to explore the role and possible mechanisms of relaxin‑3 through hydrogen peroxide (H2O2)‑induced apoptosis in primary human hepatocytes. Cells were treated with relaxin‑3 and then cell viability, morphological features, the presence of cleaved caspases as well as the levels of endoplasmic reticulum stress (ERS) protein markers and autophagy markers were evaluated. The H2O2 group showed significantly decreased cell viability, increased apoptosis as well as upregulation of caspases (cleaved caspase‑3, ‑8 and ‑9) and ERS protein markers compared with those of the control group. However, cells treated with relaxin‑3 (10 ng/ml) demonstrated improved cell viability, reduced apoptosis and decreased expression of cleaved caspases and ERS markers. However, the expression of autophagy markers remained unchanged following H2O2‑induced apoptosis and relaxin‑3 treatment. In conclusion, relaxin‑3 was shown to protect hepatocytes from H2O2‑induced apoptosis via downregulation of cleaved caspase‑8 and ‑9, as well as inhibition of the ERS pathway.

View Figures

View References

1	Horvathova E, Eckl PM, Bresgen N and Slamenova D: Evaluation of genotoxic and cytotoxic effects of H2O2 and DMNQ on freshly isolated rat hepatocytes; protective effects of carboxymethyl chitin-glucan. Neuro Endocrinol Lett. 29:644–648. 2008.PubMed/NCBI
2	Miñana JB, Gómez-Cambronero L, Lloret A, et al: Mitochondrial oxidative stress and CD95 ligand: a dual mechanism for hepatocyte apoptosis in chronic alcoholism. Hepatology. 35:1205–1214. 2002. View Article : Google Scholar : PubMed/NCBI
3	Jaeschke H, McGill MR and Ramachandran A: Oxidant stress, mitochondria, and cell death mechanisms in drug-induced liver injury: lessons learned from acetaminophen hepatotoxicity. Drug Metab Rev. 44:88–106. 2012. View Article : Google Scholar : PubMed/NCBI
4	Kundu D, Roy A, Mandal T, Bandyopadhyay U, Ghosh E and Ray D: Oxidative stress in alcoholic and viral hepatitis. N Am J Med Sci. 4:412–415. 2012. View Article : Google Scholar : PubMed/NCBI
5	Koike K: Oxidative stress and apoptosis in hepatitis C: the core issue. J Gastroenterol. 41:292–294. 2006. View Article : Google Scholar : PubMed/NCBI
6	Rodrigues CM, Fan G, Wong PY, Kren BT and Steer CJ: Ursodeoxycholic acid may inhibit deoxycholic acid-induced apoptosis by modulating mitochondrial transmembrane potential and reactive oxygen species production. Mol Med. 4:165–178. 1998.PubMed/NCBI
7	Canbay A, Gieseler RK, Gores GJ and Gerken G: The relationship between apoptosis and non-alcoholic fatty liver disease: an evolutionary cornerstone turned pathogenic. Z Gastroenterol. 43:211–217. 2005. View Article : Google Scholar : PubMed/NCBI
8	Canbay A, Friedman S and Gores GJ: Apoptosis: the nexus of liver injury and fibrosis. Hepatology. 39:273–278. 2004. View Article : Google Scholar : PubMed/NCBI
9	Matthews GM, Newbold A and Johnstone RW: Intrinsic and extrinsic apoptotic pathway signaling as determinants of histone deacetylase inhibitor antitumor activity. Adv Cancer Res. 116:165–197. 2012. View Article : Google Scholar : PubMed/NCBI
10	Breckenridge DG, Germain M, Mathai JP, Nguyen M and Shore GC: Regulation of apoptosis by endoplasmic reticulum pathways. Oncogene. 22:8608–8618. 2003. View Article : Google Scholar : PubMed/NCBI
11	McCullough KD, Martindale JL, Klotz LO, Aw TY and Holbrook NJ: Gadd153 sensitizes cells to endoplasmic reticulum stress by down-regulating Bcl2 and perturbing the cellular redox state. Mol Cell Biol. 21:1249–1259. 2001. View Article : Google Scholar : PubMed/NCBI
12	Puthalakath H, O’Reilly LA, Gunn P, Lee L, et al: ER stress triggers apoptosis by activating BH3-only protein Bim. Cell. 129:1337–1349. 2007. View Article : Google Scholar : PubMed/NCBI
13	Nakagawa T, Zhu H, Morishima N, Li E, Xu J, Yankner BA and Yuan J: Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-beta. Nature. 403:98–103. 2000. View Article : Google Scholar : PubMed/NCBI
14	Bathgate RA, Samuel CS, Burazin TC, et al: Human relaxin gene 3 (H3) and the equivalent mouse relaxin (M3) gene: Novel members of the relaxin peptide family. J Biol Chem. 277:1148–1157. 2002. View Article : Google Scholar
15	McGowan BM, Stanley SA, Ghatei MA and Bloom SR: Relaxin-3 and its role in neuroendocrine function. Ann NY Acad Sci. 1160:250–255. 2009. View Article : Google Scholar : PubMed/NCBI
16	Chan LJ, Hossain MA, Samuel CS, Separovic F and Wade JD: The relaxin peptide family - structure, function and clinical applications. Protein Pept Lett. 18:220–229. 2011. View Article : Google Scholar
17	Fallowfield JA, Hayden AL, Snowdon VK, Aucott RL, Stutchfield BM, Mole DJ, Pellicoro A, Gordon-Walker TT, Henke A, Schrader J, Trivedi PJ, Princivalle M, Forbes SJ, Collins JE and Iredale JP: Relaxin modulates human and rat hepatic myofibroblast function and ameliorates portal hypertension in vivo. Hepatology. 59:1492–1504. 2014. View Article : Google Scholar
18	Bennett RG, Dalton SR, Mahan KJ, Gentry-Nielsen MJ, Hamel FG and Tuma DJ: Relaxin receptors in hepatic stellate cells and cirrhotic liver. Biochem Pharmacol. 73:1033–1040. 2007. View Article : Google Scholar : PubMed/NCBI
19	Yao L, Agoulnik AI, Cooke PS, Meling DD and Sherwood OD: Relaxin acts on stromal cells to promote epithelial and stromal proliferation and inhibit apoptosis in the mouse cervix and vagina. Endocrinology. 149:2072–2079. 2008. View Article : Google Scholar : PubMed/NCBI
20	Moore XL, Tan SL, Lo CY, et al: Relaxin antagonizes hypertrophy and apoptosis in neonatal rat cardiomyocytes. Endocrinology. 148:1582–1589. 2007. View Article : Google Scholar : PubMed/NCBI
21	Wu CC and Bratton SB: Regulation of the intrinsic apoptosis pathway by reactive oxygen species. Antioxid Redox Signal. 19:546–558. 2013. View Article : Google Scholar :
22	Shore GC, Papa FR and Oakes SA: Signaling cell death from the endoplasmic reticulum stress response. Curr Opin Cell Biol. 23:143–149. 2011. View Article : Google Scholar :
23	Ryter SW, Cloonan SM and Choi AM: Autophagy: a critical regulator of cellular metabolism and homeostasis. Mol Cells. 36:7–16. 2013. View Article : Google Scholar : PubMed/NCBI
24	Bhogal RH, Weston CJ, Curbishley SM, Adams DH and Afford SC: Autophagy: a cyto-protective mechanism which prevents primary human hepatocyte apoptosis during oxidative stress. Autophagy. 8:545–558. 2012. View Article : Google Scholar : PubMed/NCBI
25	Tanaka M, Iijima N, Miyamoto Y, Fukusumi S, Itoh Y, Ozawa H and Ibata Y: Neurons expressing relaxin 3/INSL 7 in the nucleus incertus respond to stress. Eur J Neurosci. 21:1659–1670. 2005. View Article : Google Scholar : PubMed/NCBI
26	McGowan BM, Stanley SA, Smith KL, et al: Central relaxin-3 administration causes hyperphagia in male Wistar rats. Endocrinology. 146:3295–3300. 2005. View Article : Google Scholar : PubMed/NCBI
27	Hida T, Takahashi E, Shikata K, et al: Chronic intracerebroventricular administration of relaxin-3 increases body weight in rats. J Recept Signal Transduct Res. 26:147–158. 2006. View Article : Google Scholar : PubMed/NCBI
28	Sutton SW, Bonaventure P, Kuei C, Roland B, Chen J, Nepomuceno D, Lovenberg TW and Liu C: Distribution of G-protein-coupled receptor (GPCR)135 binding sites and receptor mRNA in the rat brain suggests a role for relaxin-3 in neuroendocrine and sensory processing. Neuroendocrinology. 80:298–307. 2004. View Article : Google Scholar
29	Smith CM, Ryan PJ, Hosken IT, Ma S and Gundlach AL: Relaxin-3 systems in the brain - the first 10 years. J Chem Neuroanat. 42:262–275. 2011. View Article : Google Scholar : PubMed/NCBI
30	Czaja MJ: Induction and regulation of hepatocyte apoptosis by oxidative stress. Antioxid Redox Signal. 4:759–767. 2002. View Article : Google Scholar : PubMed/NCBI
31	Wang H, Xue Z, Wang Q, Feng X and Shen Z: Propofol protects hepatic L02 cells from hydrogen peroxide-induced apoptosis via activation of extracellular signal-regulated kinases pathway. Anesth Analg. 107:534–540. 2008. View Article : Google Scholar : PubMed/NCBI
32	van der Westhuizen ET, Halls ML, Samuel CS, Bathgate RA, Unemori EN, Sutton SW and Summers RJ: Relaxin family peptide receptors - from orphans to therapeutic targets. Drug Discov Today. 13:640–651. 2008. View Article : Google Scholar : PubMed/NCBI
33	Zhao S, Fields PA and Sherwood OD: Evidence that relaxin inhibits apoptosis in the cervix and the vagina during the second half of pregnancy in the rat. Endocrinology. 142:2221–2229. 2001.PubMed/NCBI
34	Ghavami S, Hashemi M, Kadkhoda K, Alavian SM, Bay GH and Los M: Apoptosis in liver diseases - detection and therapeutic applications. Med Sci Monit. 11:RA337–RA345. 2005.PubMed/NCBI
35	Kannan K and Jain SK: Oxidative stress and apoptosis. Pathophysiology. 7:153–163. 2000. View Article : Google Scholar : PubMed/NCBI
36	Yin XM and Ding WX: Death receptor activation-induced hepatocyte apoptosis and liver injury. Curr Mol Med. 3:491–508. 2003. View Article : Google Scholar : PubMed/NCBI
37	Liu YY, Xie Q, Wang H, Lin LY, Jiang S, Zhou XQ, Yu H and Guo Q: The effect of N-acetyl-L-cysteine on endoplasmic reticulum stress mediated apoptosis of HepG2 cells. Zhonghua Gan Zang Bing Za Zhi. 16:524–527. 2008.(In Chinese). PubMed/NCBI
38	Loos B, Genade S, Ellis B, Lochner A and Engelbrecht AM: At the core of survival: autophagy delays the onset of both apoptotic and necrotic cell death in a model of ischemic cell injury. Exp Cell Res. 317:1437–1453. 2011. View Article : Google Scholar : PubMed/NCBI