PMID- 28330474 OWN - NLM STAT- MEDLINE DCOM- 20170515 LR - 20231213 IS - 1476-511X (Electronic) IS - 1476-511X (Linking) VI - 16 IP - 1 DP - 2017 Mar 23 TI - Hypercholesterolemia downregulates autophagy in the rat heart. PG - 60 LID - 10.1186/s12944-017-0455-0 [doi] LID - 60 AB - BACKGROUND: We have previously shown that efficiency of ischemic conditioning is diminished in hypercholesterolemia and that autophagy is necessary for cardioprotection. However, it is unknown whether isolated hypercholesterolemia disturbs autophagy or the mammalian target of rapamycin (mTOR) pathways. Therefore, we investigated whether isolated hypercholesterolemia modulates cardiac autophagy-related pathways or programmed cell death mechanisms such as apoptosis and necroptosis in rat heart. METHODS: Male Wistar rats were fed either normal chow (NORM; n = 9) or with 2% cholesterol and 0.25% cholic acid-enriched diet (CHOL; n = 9) for 12 weeks. CHOL rats exhibited a 41% increase in plasma total cholesterol level over that of NORM rats (4.09 mmol/L vs. 2.89 mmol/L) at the end of diet period. Animals were sacrificed, hearts were excised and briefly washed out. Left ventricles were snap-frozen for determination of markers of autophagy, mTOR pathway, apoptosis, and necroptosis by Western blot. RESULTS: Isolated hypercholesterolemia was associated with a significant reduction in expression of cardiac autophagy markers such as LC3-II, Beclin-1, Rubicon and RAB7 as compared to controls. Phosphorylation of ribosomal S6, a surrogate marker for mTOR activity, was increased in CHOL samples. Cleaved caspase-3, a marker of apoptosis, increased in CHOL hearts, while no difference in the expression of necroptotic marker RIP1, RIP3 and MLKL was detected between treatments. CONCLUSIONS: This is the first comprehensive analysis of autophagy and programmed cell death pathways of apoptosis and necroptosis in hearts of hypercholesterolemic rats. Our data show that isolated hypercholesterolemia suppresses basal cardiac autophagy and that the decrease in autophagy may be a result of an activated mTOR pathway. Reduced autophagy was accompanied by increased apoptosis, while cardiac necroptosis was not modulated by isolated hypercholesterolemia. Decreased basal autophagy and elevated apoptosis may be responsible for the loss of cardioprotection reported in hypercholesterolemic animals. FAU - Giricz, Zoltan AU - Giricz Z AUID- ORCID: 0000-0003-2036-8665 AD - Department of Pharmacology and Pharmacotherapy, Faculty of Medicine, Semmelweis University, Nagyvarad ter 4, H-1089, Budapest, Hungary. giricz.zoltan@med.semmelweis-univ.hu. AD - Pharmahungary Group, Szeged, Hungary. giricz.zoltan@med.semmelweis-univ.hu. FAU - Koncsos, Gabor AU - Koncsos G AD - Department of Pharmacology and Pharmacotherapy, Faculty of Medicine, Semmelweis University, Nagyvarad ter 4, H-1089, Budapest, Hungary. FAU - Rajtik, Tomas AU - Rajtik T AD - Department of Pharmacology and Toxicology, Faculty of Pharmacy, Comenius University, Odbojarov 10, 83232, Bratislava, Slovakia. FAU - Varga, Zoltan V AU - Varga ZV AD - Department of Pharmacology and Pharmacotherapy, Faculty of Medicine, Semmelweis University, Nagyvarad ter 4, H-1089, Budapest, Hungary. AD - Department of Biochemistry, Faculty of Medicine, University of Szeged, Dom ter 9, H-6720, Szeged, Hungary. FAU - Baranyai, Tamas AU - Baranyai T AD - Department of Pharmacology and Pharmacotherapy, Faculty of Medicine, Semmelweis University, Nagyvarad ter 4, H-1089, Budapest, Hungary. FAU - Csonka, Csaba AU - Csonka C AD - Department of Biochemistry, Faculty of Medicine, University of Szeged, Dom ter 9, H-6720, Szeged, Hungary. FAU - Szobi, Adrian AU - Szobi A AD - Department of Pharmacology and Toxicology, Faculty of Pharmacy, Comenius University, Odbojarov 10, 83232, Bratislava, Slovakia. FAU - Adameova, Adriana AU - Adameova A AD - Department of Pharmacology and Toxicology, Faculty of Pharmacy, Comenius University, Odbojarov 10, 83232, Bratislava, Slovakia. FAU - Gottlieb, Roberta A AU - Gottlieb RA AD - Heart Institute, Cedars-Sinai Medical Center, 127 S. San Vicente Blvd., Los Angeles, CA, 90048, USA. FAU - Ferdinandy, Peter AU - Ferdinandy P AD - Department of Pharmacology and Pharmacotherapy, Faculty of Medicine, Semmelweis University, Nagyvarad ter 4, H-1089, Budapest, Hungary. AD - Pharmahungary Group, Szeged, Hungary. LA - eng GR - P01 HL112730/HL/NHLBI NIH HHS/United States PT - Journal Article DEP - 20170323 PL - England TA - Lipids Health Dis JT - Lipids in health and disease JID - 101147696 RN - 0 (Beclin-1) RN - 0 (Becn1 protein, rat) RN - 0 (Biomarkers) RN - 0 (LC3 protein, rat) RN - 0 (Microtubule-Associated Proteins) RN - 0 (rab7 GTP-Binding Proteins) RN - 0 (rab7 GTP-binding proteins, rat) RN - 97C5T2UQ7J (Cholesterol) RN - EC 2.7.- (MLKL protein, rat) RN - EC 2.7.- (Protein Kinases) RN - EC 2.7.1.1 (mTOR protein, rat) RN - EC 2.7.11.1 (Protein Serine-Threonine Kinases) RN - EC 2.7.11.1 (RIPK1 protein, rat) RN - EC 2.7.11.1 (Receptor-Interacting Protein Serine-Threonine Kinases) RN - EC 2.7.11.1 (Ribosomal Protein S6 Kinases) RN - EC 2.7.11.1 (TOR Serine-Threonine Kinases) RN - EC 2.7.11.1 (Ripk3 protein, rat) RN - EC 3.4.22.- (Casp3 protein, rat) RN - EC 3.4.22.- (Caspase 3) RN - EC 3.6.5.2 (rab GTP-Binding Proteins) RN - G1JO7801AE (Cholic Acid) SB - IM EIN - Lipids Health Dis. 2017 Jul 5;16(1):133. PMID: 28679444 MH - Animals MH - Apoptosis/*drug effects MH - Autophagy/*drug effects MH - Beclin-1/genetics/metabolism MH - Biomarkers/metabolism MH - Caspase 3/genetics/metabolism MH - Cholesterol/administration & dosage/*adverse effects MH - Cholic Acid/administration & dosage/*adverse effects MH - Diet, High-Fat/adverse effects MH - Gene Expression Regulation/drug effects MH - Heart/drug effects MH - Hypercholesterolemia/etiology/genetics/*metabolism/pathology MH - Male MH - Microtubule-Associated Proteins/genetics/metabolism MH - Necrosis/etiology/genetics/metabolism/pathology MH - Protein Kinases/genetics/metabolism MH - Protein Serine-Threonine Kinases/genetics/metabolism MH - Rats MH - Rats, Wistar MH - Receptor-Interacting Protein Serine-Threonine Kinases/genetics/metabolism MH - Ribosomal Protein S6 Kinases/genetics/metabolism MH - Signal Transduction MH - TOR Serine-Threonine Kinases/genetics/metabolism MH - rab GTP-Binding Proteins/genetics/metabolism MH - rab7 GTP-Binding Proteins PMC - PMC5363032 OTO - NOTNLM OT - ATG8 OT - Apoptosis OT - Autophagy OT - Caspase OT - Hypercholesterolemia OT - Necroptosis OT - Programmed necrosis OT - Receptor-interacting serine/threonine-protein kinase EDAT- 2017/03/24 06:00 MHDA- 2017/05/16 06:00 PMCR- 2017/03/23 CRDT- 2017/03/24 06:00 PHST- 2016/12/16 00:00 [received] PHST- 2017/03/14 00:00 [accepted] PHST- 2017/03/24 06:00 [entrez] PHST- 2017/03/24 06:00 [pubmed] PHST- 2017/05/16 06:00 [medline] PHST- 2017/03/23 00:00 [pmc-release] AID - 10.1186/s12944-017-0455-0 [pii] AID - 455 [pii] AID - 10.1186/s12944-017-0455-0 [doi] PST - epublish SO - Lipids Health Dis. 2017 Mar 23;16(1):60. doi: 10.1186/s12944-017-0455-0.