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Aging and disease    2018, Vol. 9 Issue (6) : 1031-1042     DOI: 10.14336/AD.2018.0221
Orginal Article |
Omi/HtrA2 Participates in Age-Related Autophagic Deficiency in Rat Liver
Xu Jiahui, Jiao Kun, Liu Xin, Sun Qi, Wang Ke, Xu Haibo, Zhang Shangyue, Wu Ye, Wu Linguo, Liu Dan, Wang Wen*, Liu Huirong*
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, and Beijing Key Laboratory of Metabolic Disorders Related Cardiovascular Diseases, Capital Medical University, Beijing, China.
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Liver is a vital organ with many important functions, and the maintenance of normal hepatic function is necessary for health. As an essential mechanism for maintaining cellular homeostasis, autophagy plays an important role in ensuring normal organ function. Studies have indicated that the degeneration of hepatic function is associated with autophagic deficiency in aging liver. However, the underlying mechanisms still remain unclear. The serine protease Omi/HtrA2 belongs to the HtrA family and promotes apoptosis through either the caspase-dependent or caspase-independent pathway. Mice lacking Omi/HtrA2 exhibited progeria symptoms (premature aging), which were similar to the characteristics of autophagic insufficiency. In this study, we demonstrated that both the protein level of Omi/HtrA2 in liver and hepatic function were reduced as rats aged, and there was a positive correlation between them. Furthermore, several autophagy-related proteins (LC3II/I, Beclin-1 and LAMP2) in rat liver were decreased significantly with the increasing of age. Finally, inhibition of Omi/HtrA2 resulted in reduced autophagy and hepatic dysfunction. In conclusion, these results suggest that Omi/HtrA2 participates in age-related autophagic deficiency in rat liver. This study may offer a novel insight into the mechanism involved in liver aging.

Keywords Omi/HtrA2      age      autophagy      liver     
Corresponding Authors: Wang Wen,Liu Huirong   
About author:

Jiahui Xu and Kun Jiao contributed equally to this work.

Issue Date: 25 November 2017
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Xu Jiahui
Jiao Kun
Liu Xin
Sun Qi
Wang Ke
Xu Haibo
Zhang Shangyue
Wu Ye
Wu Linguo
Liu Dan
Wang Wen
Liu Huirong
Cite this article:   
Xu Jiahui,Jiao Kun,Liu Xin, et al. Omi/HtrA2 Participates in Age-Related Autophagic Deficiency in Rat Liver[J]. Aging and disease, 2018, 9(6): 1031-1042.
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Figure 1.  Both the level of aging-relative proteins and oxidative stress were increased in natural aging rat models. (A-D) The results of immunostaining and immunoblotting indicated that the expression of p53, p21 and β-gal was increased with growth of age. **P<0.01vs. 3 months, ##P<0.01 vs.9 month, #P<0.05 vs.9-month, n=6-8. (E-F) Compared with 3 and 9-month groups, there were decreased SOD activity and increased MDA level in 22-month group. **P<0.01vs. 3 months, #P<0.05 vs.9-month, n=6-8. SOD, Superoxide dismutase; MDA, Malondialdehyde.
Figure 2.  The morphology and function of liver were declined during the natural aging process. (A) The liver morphology of 3, 9, 22-month-old rats was detected by H&E staining, bar=50 μm. (B) The hepatic function was evaluated by serum biochemical detection. (C-F) With the growth of age, both AST and GLB were increased significantly, the level of ALB and the ALB/GLB ratio were decreased. **P<0.01vs. 3 months, #P<0.05 vs.9 month, &P<0.05 vs.3 month. n=8-10. ALB, albumin; GLB, globin; AST, aspartate aminotransferase.
Figure 3.  Decreasing of Omi/HtrA2 was accompanied with hepatic dysfunction in the natural aging process. (A) The protein level of Omi/HtrA2 in the liver of 3, 9, 22-month-old rats. Immunostaining, bar=200 μm. (B) Immunoblotting results indicated that the level of Omi/HtrA2 was decreased with growth of age. **P<0.01vs. 3 months, #P<0.05 vs.9 month. n=6-8. (C) The mRNA level of Omi/HtrA2 was detected by quantitative RT-PCR. (D-H) Correlation analysis of Omi/HtrA2 and hepatic function. There was a positive correlation between the protein level of Omi/HtrA2 in liver and the index of hepatic function. ALB, albumin; GLB, globin; AST, aspartate aminotransferase; ALT, alanine aminotransferase.
Figure 4.  The level of autophagy in liver was decreased with aging. (A) The immunostaining of LC3II, bar=100 μm. (B) Immunoblotting results indicated that the level of autophagy was decreased with growth of age. **P<0.01vs. 3 months, #P<0.05 vs.9 month. n=6-8.
Figure 5.  Inhibition of Omi/HtrA2 resulted in reduced autophagy in liver. (A-B) After treatment with ucf-101, the autophagy-related protein levels in 3 and 9-month-old rat’s liver were detected by immunoblotting. *P<0.05 vs. 3-month control, &P<0.05 vs. 9-month control. n=8-10. (C-D) The eGFP-mRFP-LC3 assays in vitro. The autophagy flux was reduced when cells were treated with ucf-101 (9.5μmol/L, 48h). Representative images showed LC3 staining in different groups of 7701 cells infected with GFP-RFP-LC3 adenovirus for 24 h. Acidified autophagosomes (red arrowheads in merged images) indicated active flux, yellow arrowheads pointed to immature autophagosomes. bar=25 μm. *P<0.05, #P<0.05. n=8. CQ, chloroquine.
Figure 6.  Inhibition of Omi/HtrA2 attenuated hepatic function. The effect of Omi/HtrA2 inhibitor ucf-101 on hepatic function from different rats was evaluated by serum biochemical detection. *P<0.05, **P<0.01. n=8-10.
[1] Lipinska B, Fayet O, Baird L, Georgopoulos C (1989). Identification, characterization, and mapping of the Escherichia coli htrA gene, whose product is essential for bacterial growth only at elevated temperatures. J Bacteriol, 171:1574–84.
[2] Faccio L, Fusco C, Chen A, Martinotti S, Bonventre JV, Zervos AS (2000). Characterization of a novel human serine protease that has extensive homology to bacterial heat shock endoprotease HtrA and is regulated by kidney ischemia. J Biol Chem, 275(4):2581–8.
[3] Hegde R, Srinivasula SM, Zhang Z, Wassell R, Mukattash R, Cilenti L, et al. (2002). Identification of Omi/HtrA2 as a mitochondrial apoptotic serine protease that disrupts inhibitor of apoptosis protein-caspase interaction. J Biol Chem, 277: 432–438.
[4] Dagda RK, Chu CT (2009). Mitochondrial quality control: insights on how Parkinson's disease related genes PINK1, parkin, and Omi/HtrA2 interact to maintain mitochondrial homeostasis. J Bioenerg Biomembr, 41:473–9.
[5] Jones JM, Datta P, Srinivasula SM, Ji W, Gupta S, Zhang Z, et al. (2003). Loss of Omi mitochondrial protease activity causes the neuromuscular disorder of mnd2 mutant mice. Nature, 425: 721–727.
[6] Kang S, Louboutin JP, Datta P, Landel CP, Martinez D, Zervos AS, et al. (2013). Loss of HtrA2/Omi activity in non-neuronal tissues of adult mice causes premature aging. Cell Death Differ, 20: 259–269.
[7] Kang S, Fernandes-Alnemri T, Alnemri ES (2013). A novel role for the mitochondrial HTRA2/OMI protease in aging. Autophagy, 9: 420–421.
[8] Strauss KM, Martins LM, Plun-Favreau H, Marx FP, Kautzmann S, Berg D, et al. (2005). Loss offunction mutations in the gene encoding Omi/HtrA2 in Parkinson’s disease. Hum MolGenet, 14: 2099–2111.
[9] Martins LM, Morrison A, Klupsch K, Fedele V, Moisoi N, Teismann P, et al. (2004). Neuroprotective role of the Reaper-related serine protease HtrA2/Omi revealed by targeted deletion in mice. Mol Cell Biol, 24: 9848–9862.
[10] Wagner KH, Cameron-Smith D, Wessner B, Franzke B (2016). Biomarkers of Aging: From Function to Molecular Biology. Nutrients, 8: 338.
[11] Anantharaju A, Feller A, Chedid A (2002). Aging Liver. A review. Gerontology, 48: 343–353.
[12] Liu D, Liu X, Wu Y, Wang W, Ma X, Liu H (2016). Cloning and Transcriptional Activity of the mouse Omi/HtrA2 gene promoter. Int J Mol Sci, 17: 119.
[13] Mizushima N, Yoshimori T, Levine B (2010). Methods in mammalian autophagy research. Cell, 140: 313–326.
[14] Galluzzi L, Pietrocola F, Levine B, Kroemer G (2014). Metabolic control of autophagy. Cell, 159: 1263–1276.
[15] Martinez-Lopez N, Athonvarangkul D, Singh R (2015). Autophagy and aging. Adv Exp Med Biol, 847: 73–87.
[16] Schneider JL, Cuervo AM (2014). Liver autophagy: much more than just taking out the trash. Nat Rev Gastroenterol Hepatol, 11: 187–200.
[17] Uddin MN, Nishio N, Ito S, Suzuki H, Isobe K (2012). Autophagic activity in thymus and liver during aging. Age (Dordr), 34: 75–85.
[18] Zhang C, Cuervo AM (2008). Restoration of chaperone-mediated autophagy in aging liver improves cellular maintenance and hepatic function. Nat Med, 14: 959–965.
[19] Rautou PE, Mansouri A, Lebrec D, Durand F, Valla D, Moreau R (2010). Autophagy in liver diseases. J Hepatol, 53: 1123–1134.
[20] Donati A, Cavallini G, Paradiso C, Vittorini S, Pollera M, Gori Z, et al. (2001). Age-related changes in the regulation of autophagic proteolysis in rat isolated hepatocytes. J Gerontol a Biol Sci Med Sci, 56: B288–B293.
[21] Li B, Hu Q, Wang H, Man N, Ren H, Wen L, et al. (2010). Omi/HtrA2 is a positive regulator of autophagy that facilitates the degradation of mutant proteins involved in neurodegenerative diseases. Cell Death Differ, 17: 1773–1784.
[22] Cilenti L, Lee Y, Hess S, Srinivasula S, Park KM, Junqueira D, et al. (2003). Characterization of a novel and specific inhibitor for the pro-apoptotic protease Omi/HtrA2. J Biol Chem, 278: 11489–11494.
[23] Liu HR, Gao E, Hu A, Tao L, Qu Y, Most P, et al. (2005). Role of Omi/HtrA2 in apoptotic cell death after myocardial ischemia and reperfusion. Circulation, 111: 90–96.
[24] Hua F, Li K, Yu JJ, et al. (2015). TRB3 links insulin/IGF to tumour promotion by interacting with p62 and impeding autophagic/proteasomal degradations. Nat Commun, 6:7951
[25] Mizushima N, Yoshimorim T, Levine B, et al. (2010). Methods in Mammalian Autophagy Research. Cell, 140(3): 313–326.
[26] Hariharan N, Zhai P, Sadoshima J (2011). Oxidative stress stimulates autophagic flux during ischemia/reperfusion. Antioxid Redox Signal, 14(11): 2179–2190.
[27] Alkan A, Koksoy EB, Utkan G (2015). Albumin to globulin ratio, a predictor or a misleader? Ann Oncol, 26: 443–444.
[28] Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, et al. (2000). LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. Embo J, 19: 5720–5728.
[29] Kang R, Zeh HJ, Lotze MT, Tang D (2011). The Beclin 1 network regulates autophagy and apoptosis. Cell Death Differ, 18: 571–580.
[30] Maejima Y, Isobe M, Sadoshima J (2016). Regulation of autophagy by Beclin 1 in the heart. J Mol Cell Cardiol, 95: 19–25.
[31] Eskelinen EL (2006). Roles of LAMP-1 and LAMP-2 in lysosome biogenesis and autophagy. Mol Aspects Med, 27: 495–502.
[32] Clausen T, Southan C, Ehrmann M (2002). The HtrA family of proteases: implications for protein composition and cell fate. Mol Cell, 10: 443–455.
[33] Suzuki Y, Imai Y, Nakayama H, Takahashi K, Takio K, Takahashi R (2001). A serine protease, HtrA2, is released from the mitochondria and interacts with XIAP, inducing cell death. Mol Cell, 8: 613–621.
[34] Martins LM, Iaccarino I, Tenev T, Gschmeissner S, Totty NF, Lemoine NR, et al. (2002). The serine protease Omi/HtrA2 regulates apoptosis by binding XIAP through a reaper-like motif. J Biol Chem, 277: 439–444.
[35] Goo HG, Rhim H, Kang S (2014). HtrA2/Omi influences the stability of LON protease 1 and prohibitin, proteins involved in mitochondrial homeostasis. Exp Cell Res, 328: 456–465.
[36] Watanabe T, Asaka S, Kitagawa D, Saito K, Kurashige R, Sasado T, et al. (2004). Mutations affecting liver development and function in Medaka, Oryzias latipes, screened by multiple criteria. Mech Dev, 121: 791–802.
[37] Gewirtz DA (2013). Autophagy and senescence: a partnership in search of definition. Autophagy, 9: 808–812.
[38] Czaja MJ, Ding WX, Donohue TJ, Friedman SL, Kim JS, Komatsu M, et al. (2013). Functions of autophagy in normal and diseased liver. Autophagy, 9: 1131–1158.
[39] Feng Y, He D, Yao Z, Klionsky DJ (2014). The machinery of macroautophagy. Cell Res, 24: 24–41.
[40] Gelino S, Hansen M (2012). Autophagy - An Emerging Anti-Aging Mechanism. J Clin Exp Pathol, Suppl 4. Pii 006.
[41] Zhou H, Chen J, Lu X, Shen C, Zeng J, Chen L, Pei Z (2012). Melatonin protects against rotenone-induced cell injury via inhibition of Omi and Bax-mediated autophagy in Hela cells. J Pineal Res, 52(1):120–7.
[42] Cilenti L, Ambivero CT, Ward N, Alnemri ES, Germain D, Zervos AS. (2014). Inactivation of Omi/HtrA2 protease leads to the deregulation of mitochondrial Mulan E3 ubiquitin ligase and increased mitophagy. Biochim Biophys Acta, 1843(7): 1295–1307
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