title: Recombinant adeno-associated virus-delivered hypoxia-inducible stanniocalcin-1 expression effectively inhibits hypoxia-induced cell apoptosis in cardiomyocytes.
abstract: Ischemia/hypoxia-induced oxidative stress is detrimental for the survival of cardiomyocytes and cardiac function. Stanniocalcin-1 (STC-1), a glycoprotein, has been found to play an inhibitory role in the production of reactive oxygen species (ROS). Here, we speculated that the overexpression of STC-1 might alleviate oxidative damage in cardiomyocytes under conditions of hypoxia. To control the expression of STC-1 in hypoxia, we constructed a recombinant adeno-associated virus (AAV) carrying the hypoxia-responsive element (HRE) to mediate hypoxia induction. Cardiomyocytes were infected with AAV-HRE-STC-1 and cultured in normoxic or hypoxic conditions, and STC-1 overexpression was only detected in hypoxic cultured cardiomyocytes by using quantitative real-time polymerase chain reaction and Western blot analysis. Using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, AAV-HRE-STC-1 infection was shown to significantly enhance cell survival under hypoxia. Hypoxia-induced cell apoptosis was inhibited by AAV-HRE-STC-1 infection by using the Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide apoptosis assay. Moreover, the proapoptotic protein Caspase-3 and anti-apoptotic protein Bcl-2, which were dysregulated by hypoxia, were reversed by AAV-HRE-STC-1 infection. AAV-HRE-STC-1-mediated STC-1 overexpression markedly inhibited ROS production in cardiomyocytes cultured under hypoxic conditions. AAV-HRE-STC-1 infection significantly upregulated uncoupled protein 3 (UCP3), whereas silencing of UCP3 blocked the inhibitory effect of AAV-HRE-STC-1 on ROS production. In contrast, AAV-HRE-STC-1 infection had no effect on UCP2, and knockdown of UCP2 did not block the inhibitory effect of AAV-HRE-STC-1 on ROS production in the cardiomyocytes cultured under hypoxic conditions. Taken together, STC1 activates antioxidant pathway in cardiomyocytes through the induction of UCP3, implying that AAV-HRE-STC-1 has potential in the treatment of ischemic-related heart disease.
authors.affiliation: *Department of Internal Medicine, The First Affiliated Hospital of Henan University of Traditional Chinese Medicine (TCM), Zhengzhou, China; †Department of Cardiology, Zhengzhou Hospital of Traditional Chinese Medicine, Zhengzhou, China; and ‡Department of Functional Examination, The First Affiliated Hospital of Henan University of Traditional Chinese Medicine (TCM), Zhengzhou, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Shi
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Qin
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D015687:Cell Hypoxia; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics; D000229:Dependovirus / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors / Q000008:administration & dosage; D006023:Glycoproteins / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D007473:Ion Channels / Q000235:genetics; D024101:Mitochondrial Proteins / Q000235:genetics; D032383:Myocytes, Cardiac / Q000473:pathology*; D018384:Oxidative Stress / Q000235:genetics; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D020218:Response Elements / Q000235:genetics; D000071247:Uncoupling Protein 3; D015854:Up-Regulation
chemical_list: D006023:Glycoproteins; D007473:Ion Channels; D024101:Mitochondrial Proteins; D017382:Reactive Oxygen Species; C000604392:UCP3 protein, human; C000604394:Ucp3 protein, rat; D000071247:Uncoupling Protein 3; C051795:teleocalcin