title: 5-aminolevulinic acid combined with sodium ferrous citrate ameliorates H2O2-induced cardiomyocyte hypertrophy via activation of the MAPK/Nrf2/HO-1 pathway.
abstract: Hydrogen peroxide (H2O2) causes cell damage via oxidative stress. Heme oxygenase-1 (HO-1) is an antioxidant enzyme that can protect cardiomyocytes against oxidative stress. In this study, we investigated whether the heme precursor 5-aminolevulinic acid (5-ALA) with sodium ferrous citrate (SFC) could protect cardiomyocytes from H2O2-induced hypertrophy via modulation of HO-1 expression. HL-1 cells pretreated with/without 5-ALA and SFC were exposed to H2O2 to induce a cardiomyocyte hypertrophy model. Hypertrophy was evaluated by planar morphometry, (3)H-leucine incorporation, and RT-PCR analysis of hypertrophy-related gene expressions. Reactive oxygen species (ROS) production was assessed by 5/6-chloromethyl-2',7'-ichlorodihydrofluorescein diacetate acetylester. HO-1 and nuclear factor erythroid 2-related factor 2 (Nrf2) protein expressions were analyzed by Western blot. In our experiments, HL-1 cells were transfected with Nrf2 siRNA or treated with a signal pathway inhibitor. We found several results. 1) ROS production, cell surface area, protein synthesis, and expressions of hypertrophic marker genes, including atrial natriuretic peptide, brain natriuretic peptide, atrial natriuretic factor, and β-myosin heavy chain, were decreased in HL-1 cells pretreated with 5-ALA and SFC. 2) 5-ALA and SFC increased HO-1 expression in a dose- and time-dependent manner, associated with upregulation of Nrf2. Notably, Nrf2 siRNA dramatically reduced HO-1 expression in HL-1 cells. 3) ERK1/2, p38, and SAPK/JNK signaling pathways were activated and modulate 5-ALA- and SFC-enhanced HO-1 expression. SB203580 (p38 kinase), PD98059 (ERK), or SP600125 (JNK) inhibitors significantly reduced this effect. In conclusion, our data suggest that 5-ALA and SFC protect HL-1 cells from H2O2-induced cardiac hypertrophy via activation of the MAPK/Nrf2/HO-1 signaling pathway.
authors: Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China;
authors: Mingyi
authors: M
authors: Zhao
authors: Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China;
authors: Huiming
authors: H
authors: Guo
authors: Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China;
authors: Jimei
authors: J
authors: Chen
authors: M
authors: M
authors: Fujino
authors: H
authors: H
authors: Ito
authors: K
authors: K
authors: Takahashi
authors: F
authors: F
authors: Abe
authors: M
authors: M
authors: Nakajima
authors: T
authors: T
authors: Tanaka
authors: Department of Pharmacology and Toxicology, Boonshoft School of Medicine, Wright State University, Dayton, Ohio;
authors: Jinju
authors: J
authors: Wang
authors: Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China;
authors: Huanlei
authors: H
authors: Huang
authors: Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China;
authors: Shaoyi
authors: S
authors: Zheng
authors: Department of Pediatrics, the Third Xiangya Hospital, Central South University, Changsha, China;
authors: Mingyan
authors: M
authors: Hei
authors: Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China;
authors: Jiaxin
authors: J
authors: Li
authors: Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China;
authors: Shuai
authors: S
authors: Huang
authors: Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China;
authors: Jiani
authors: J
authors: Li
authors: Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases, Affiliated Hospital of Guangdong Medical College, Zhanjiang, China.
authors: Xiaotang
authors: X
authors: Ma
authors: Department of Pharmacology and Toxicology, Boonshoft School of Medicine, Wright State University, Dayton, Ohio;
authors: Yanfang
authors: Y
authors: Chen
authors: Department of Pediatrics, the Third Xiangya Hospital, Central South University, Changsha, China;
authors: Lingling
authors: L
authors: Zhao
authors: Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China;
authors: Jian
authors: J
authors: Zhuang
authors: Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, China; zhupingzggd@126.com.
authors: Ping
authors: P
authors: Zhu
authors: X K
authors: XK
authors: Li
mesh_terms: D000622:Aminolevulinic Acid / Q000494:pharmacology*; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D009320:Atrial Natriuretic Factor / Q000378:metabolism; D006332:Cardiomegaly / Q000188:drug therapy* / Q000473:pathology; D002460:Cell Line; D019343:Citric Acid; D005296:Ferrous Compounds / Q000494:pharmacology*; D051547:Heme Oxygenase-1 / Q000096:biosynthesis / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008565:Membrane Proteins / Q000096:biosynthesis / Q000378:metabolism; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000473:pathology*; D018995:Myosin Heavy Chains / Q000378:metabolism; D051267:NF-E2-Related Factor 2 / Q000235:genetics* / Q000378:metabolism; D020097:Natriuretic Peptide, Brain / Q000378:metabolism; D018384:Oxidative Stress; D034622:RNA Interference; D034741:RNA, Small Interfering; D015398:Signal Transduction; D024723:Ventricular Myosins / Q000378:metabolism
chemical_list: D000975:Antioxidants; D005296:Ferrous Compounds; D008565:Membrane Proteins; D051267:NF-E2-Related Factor 2; C495636:Nfe2l2 protein, mouse; D034741:RNA, Small Interfering; D020097:Natriuretic Peptide, Brain; D019343:Citric Acid; C016600:ferrous citrate; D009320:Atrial Natriuretic Factor; D000622:Aminolevulinic Acid; D006861:Hydrogen Peroxide; D051547:Heme Oxygenase-1; C496542:Hmox1 protein, mouse; D020928:Mitogen-Activated Protein Kinases; D024723:Ventricular Myosins; D018995:Myosin Heavy Chains
keywords: 5-aminolevulinic acid; cardiomyocyte hypertrophy; heme oxygenase-1; mitogen-activated protein kinase; nuclear factor erythroid 2-related factor 2