PPARδ activation protects H9c2 cardiomyoblasts from LPS‑induced apoptosis through the heme oxygenase‑1‑mediated suppression of NF‑κB activation

Shi,Yao; Jiang,Hong; Yang,Xiaobo

doi:10.3892/mmr.2017.6483

PPARδ activation protects H9c2 cardiomyoblasts from LPS‑induced apoptosis through the heme oxygenase‑1‑mediated suppression of NF‑κB activation

Authors:
- Yao Shi
- Hong Jiang
- Xiaobo Yang
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Affiliations: Department of Neonatology, the Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430014, P.R. China, Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P.R. China
Published online on: April 19, 2017 https://doi.org/10.3892/mmr.2017.6483
Pages: 3775-3780

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Abstract

The aim of the present study was to investigate the protective effect of the selective peroxisome proliferator-activated receptor δ (PPARδ) agonist GW501516 (GW) on lipopolysaccharide (LPS)‑induced apoptosis in the rat cardiomyoblast cell line H9c2, and to investigate the possible underlying mechanisms. Cell viability was estimated using the MTT assay. Apoptosis was estimated by flow cytometry using Annexin V‑fluorescein isothiocyanate/propidium iodide staining and caspase‑3 activity assay. The protein level of heme oxygenase‑1 (HO‑1), cleaved caspase‑3 (CC3), apoptosis regulator Bcl‑2 (bcl‑2), apoptosis regulator BAX (bax) and nuclear factor‑κB (NF‑κB) p65 was measured by western blot analysis. The results demonstrated that pretreatment with GW inhibited the LPS‑induced increase in the rate of apoptosis. Pretreatment with GW also increased the bcl‑2/bax ratio, and decreased CC3 protein expression as well as caspase‑3 activity, in LPS‑stimulated H9c2 cells. Further studies demonstrated that GW inhibited LPS‑induced NF‑κB nuclear translocation in a dose‑dependent manner. In addition, GW induced HO‑1 protein expression in a dose‑dependent manner. ZnPP‑IX, an inhibitor of HO‑1, reversed the inhibitory effect of GW on LPS‑induced NF‑κB activation, leading to the attenuation of PPARδ‑mediated apoptosis resistance. In conclusion, these results suggest that PPARδ activation exerts an anti‑apoptotic effect in LPS‑stimulated H9c2 cardiomyoblasts, potentially through heme oxygenase‑1‑mediated suppression of NF‑κB activation. PPARδ appears to be a promising therapeutic target for the treatment of sepsis‑associated cardiac dysfunction.

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1	Czaja AS, Zimmerman JJ and Nathens AB: Readmission and late mortality after pediatric severe sepsis. Pediatrics. 123:849–857. 2009. View Article : Google Scholar : PubMed/NCBI
2	Watson RS, Carcillo JA, Linde-Zwirble WT, Clermont G, Lidicker J and Angus DC: The epidemiology of severe sepsis in children in the United States. Am J Respir Crit Care Med. 167:695–701. 2003. View Article : Google Scholar : PubMed/NCBI
3	Maeder M, Fehr T, Rickli H and Ammann P: Sepsis-associated myocardial dysfunction: Diagnostic and prognostic impact of cardiac troponins and natriuretic peptides. Chest. 129:1349–1366. 2006. View Article : Google Scholar : PubMed/NCBI
4	Ayala A, Perl M, Venet F, Lomas-Neira J, Swan R and Chung CS: Apoptosis in sepsis: Mechanisms, clinical impact and potential therapeutic targets. Curr Pharm Des. 14:1853–1859. 2008. View Article : Google Scholar : PubMed/NCBI
5	Ehrenborg E and Krook A: Regulation of skeletal muscle physiology and metabolism by peroxisome proliferator-activated receptor delta. Pharmacol Rev. 61:373–393. 2009. View Article : Google Scholar : PubMed/NCBI
6	Barish GD, Narkar VA and Evans RM: PPAR delta: A dagger in the heart of the metabolic syndrome. J Clin Invest. 116:590–597. 2006. View Article : Google Scholar : PubMed/NCBI
7	Li AC, Binder CJ, Gutierrez A, Brown KK, Plotkin CR, Pattison JW, Valledor AF, Davis RA, Willson TM, Witztum JL, et al: Differential inhibition of macrophage foam-cell formation and atherosclerosis in mice by PPARalpha, beta/delta, and gamma. J Clin Invest. 114:1564–1576. 2004. View Article : Google Scholar : PubMed/NCBI
8	Liou JY, Lee S, Ghelani D, Matijevic-Aleksic N and Wu KK: Protection of endothelial survival by peroxisome proliferator-activated receptor-delta mediated 14-3-3 upregulation. Arterioscler Thromb Vasc Biol. 26:1481–1487. 2006. View Article : Google Scholar : PubMed/NCBI
9	Piqueras L, Reynolds AR, Hodivala-Dilke KM, Alfranca A, Redondo JM, Hatae T, Tanabe T, Warner TD and Bishop-Bailey D: Activation of PPARbeta/delta induces endothelial cell proliferation and angiogenesis. Arterioscler Thromb Vasc Biol. 27:63–69. 2007. View Article : Google Scholar : PubMed/NCBI
10	Pesant M, Sueur S, Dutartre P, Tallandier M, Grimaldi PA, Rochette L and Connat JL: Peroxisome proliferator-activated receptor delta (PPARdelta) activation protects H9c2 cardiomyoblasts from oxidative stress-induced apoptosis. Cardiovasc Res. 69:440–449. 2006. View Article : Google Scholar : PubMed/NCBI
11	Carlson D, Maass DL, White DJ, Tan J and Horton JW: Antioxidant vitamin therapy alters sepsis-related apoptotic myocardial activity and inflammatory responses. Am J Physiol Heart Circ Physiol. 291:H2779–H2789. 2006. View Article : Google Scholar : PubMed/NCBI
12	Otterbein LE and Choi AM: Heme oxygenase: Colors of defense against cellular stress. Am J Physiol Lung Cell Mol Physiol. 279:L1029–L1037. 2000.PubMed/NCBI
13	Ali F, Ali NS, Bauer A, Boyle JJ, Hamdulay SS, Haskard DO, Randi AM and Mason JC: PPARdelta and PGC1alpha act cooperatively to induce haem oxygenase-1 and enhance vascular endothelial cell resistance to stress. Cardiovasc Res. 85:701–710. 2010. View Article : Google Scholar : PubMed/NCBI
14	Zabalgoitia M, Colston JT, Reddy SV, Holt JW, Regan RF, Stec DE, Rimoldi JM, Valente AJ and Chandrasekar B: Carbon monoxide donors or heme oxygenase-1 (HO-1) overexpression blocks interleukin-18-mediated NF-kappaB-PTEN-dependent human cardiac endothelial cell death. Free Radic Biol Med. 44:284–298. 2008. View Article : Google Scholar : PubMed/NCBI
15	Tien YC, Lin JY, Lai CH, Kuo CH, Lin WY, Tsai CH, Tsai FJ, Cheng YC, Peng WH and Huang CY: Carthamus tinctorius L. Prevents LPS-induced TNFalpha signaling activation and cell apoptosis through JNK1/2-NFkappaB pathway inhibition in H9c2 cardiomyoblast cells. J Ethnopharmacol. 130:505–513. 2010. View Article : Google Scholar : PubMed/NCBI
16	Jiang B, Liang P, Zhang B, Song J, Huang X and Xiao X: Role of PPAR-beta in hydrogen peroxide-induced apoptosis in human umbilical vein endothelial cells. Atherosclerosis. 204:353–358. 2009. View Article : Google Scholar : PubMed/NCBI
17	Cryns V and Yuan J: Proteases to die for. Genes Dev. 12:1551–1570. 1998. View Article : Google Scholar : PubMed/NCBI
18	Oltvai ZN, Milliman CL and Korsmeyer SJ: Bcl-2 heterodimerizes in vivo with a conserved homolog, Bax, that accelerates programmed cell death. Cell. 74:609–619. 1993. View Article : Google Scholar : PubMed/NCBI
19	Wang X, Zingarelli B, O'Connor M, Zhang P, Adeyemo A, Kranias EG, Wang Y and Fan GC: Overexpression of Hsp20 prevents endotoxin-induced myocardial dysfunction and apoptosis via inhibition of NF-kappaB activation. J Mol Cell Cardiol. 47:382–390. 2009. View Article : Google Scholar : PubMed/NCBI
20	Zingarelli B, Piraino G, Hake PW, O'Connor M, Denenberg A, Fan H and Cook JA: Peroxisome proliferator-activated receptor {delta} regulates inflammation via NF-{kappa}B signaling in polymicrobial sepsis. Am J Pathol. 177:1834–1847. 2010. View Article : Google Scholar : PubMed/NCBI
21	Abdulkhalek S, Guo M, Amith SR, Jayanth P and Szewczuk MR: G-protein coupled receptor agonists mediate Neu1 sialidase and matrix metalloproteinase-9 cross-talk to induce transactivation of TOLL-like receptors and cellular signaling. Cell Signal. 24:2035–2042. 2012. View Article : Google Scholar : PubMed/NCBI
22	Delhalle S, Blasius R, Dicato M and Diederich M: A beginner's guide to NF-kappaB signaling pathways. Ann N Y Acad Sci. 1030:1–13. 2004. View Article : Google Scholar : PubMed/NCBI
23	Wang G, Hamid T, Keith RJ, Zhou G, Partridge CR, Xiang X, Kingery JR, Lewis RK, Li Q, Rokosh DG, et al: Cardioprotective and antiapoptotic effects of heme oxygenase-1 in the failing heart. Circulation. 121:1912–1925. 2010. View Article : Google Scholar : PubMed/NCBI
24	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
25	Kim HJ, Ham SA, Paek KS, Hwang JS, Jung SY, Kim MY, Jin H, Kang ES, Woo IS, Kim HJ, et al: Transcriptional up-regulation of antioxidant genes by PPARδ inhibits angiotensin II-induced premature senescence in vascular smooth muscle cells. Biochem Biophys Res Commun. 406:564–569. 2011. View Article : Google Scholar : PubMed/NCBI
26	Bernardini C, Zannoni A, Bacci ML and Forni M: Protective effect of carbon monoxide pre-conditioning on LPS-induced endothelial cell stress. Cell Stress Chaperones. 15:219–224. 2010. View Article : Google Scholar : PubMed/NCBI