title: SIRT3 positively regulates the expression of folliculogenesis- and luteinization-related genes and progesterone secretion by manipulating oxidative stress in human luteinized granulosa cells.
abstract: SIRT3 is a member of the sirtuin family and has recently emerged as a vital molecule in controlling the generation of reactive oxygen species (ROS) in oocytes. Appropriate levels of ROS play pivotal roles in human reproductive medicine. The aim of the present study was to investigate SIRT3 expression and analyze the SIRT3-mediated oxidative response in human luteinized granulosa cells (GCs). Human ovarian tissues were subjected to immunohistochemical analysis to localize SIRT3 expression. Hydrogen peroxide and human chorionic gonadotropin were used to analyze the relationship between ROS and SIRT3 by quantitative RT-PCR and Western blotting. Intracellular levels of ROS were investigated by fluorescence after small interfering RNA-mediated knockdown of SIRT3 in human GCs. To uncover the role of SIRT3 in folliculogenesis and luteinization, mRNA levels of related genes and the progesterone concentration were analyzed by quantitative RT-PCR and immunoassays, respectively. We detected the expression of SIRT3 in the GCs of the human ovary. The mRNA levels of SIRT3, catalase, and superoxide dismutase 1 were up-regulated by hydrogen peroxide in both COV434 cells and human GCs and down-regulated by human chorionic gonadotropin. Knockdown of SIRT3 markedly elevated ROS generation in human GCs. In addition, SIRT3 depletion resulted in decreased mRNA expression of aromatase, 17β-hydroxysteroid dehydrogenase 1, steroidogenic acute regulatory protein, cholesterol side-chain cleavage enzyme, and 3β-hydroxysteroid dehydrogenase in GCs and thus resulted in decreased progesterone secretion. These results have the important clinical implication that SIRT3 might play a positive role in the folliculogenesis and luteinization processes in GCs, possibly by sensing and regulating the generation of ROS. Activation of SIRT3 function might help to sustain human reproduction by maintaining GCs as well as oocytes.
authors.affiliation: Department of Obstetrics and Gynecology (H.F., O.W.-H., M.H., A.Sa., A.Sh., Y.M., W.I., K.Ko., K.O., K.Ka. Y.O., T.F.) and the University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan; Department of Physiological Chemistry and Metabolism (Y.K., W.I., H.K.), Graduate School of Medicine, the University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan; Department of Obstetrics and Gynecology (H.O., T.Y.), National Center for Global Health and Medicine, 1-21-1 Toyama, Shinjyuku-ku, Tokyo 162-8655, Japan.
authors.forename: Houju
authors.initials: H
authors.lastname: Fu
authors.forename: Osamu
authors.initials: O
authors.lastname: Wada-Hiraike
authors.forename: Mana
authors.initials: M
authors.lastname: Hirano
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Kawamura
authors.forename: Ayako
authors.initials: A
authors.lastname: Sakurabashi
authors.forename: Akira
authors.initials: A
authors.lastname: Shirane
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Morita
authors.forename: Wataru
authors.initials: W
authors.lastname: Isono
authors.forename: Hajime
authors.initials: H
authors.lastname: Oishi
authors.forename: Kaori
authors.initials: K
authors.lastname: Koga
authors.forename: Katsutoshi
authors.initials: K
authors.lastname: Oda
authors.forename: Kei
authors.initials: K
authors.lastname: Kawana
authors.forename: Tetsu
authors.initials: T
authors.lastname: Yano
authors.forename: Hiroki
authors.initials: H
authors.lastname: Kurihara
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Osuga
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Fujii
mesh_terms: D000328:Adult; D000975:Antioxidants / Q000378:metabolism; D002374:Catalase / Q000378:metabolism; D045744:Cell Line, Tumor; D006063:Chorionic Gonadotropin; D005260:Female; D006107:Granulosa Cells / Q000378:metabolism*; D006801:Humans; D034461:Luteinization; D006080:Ovarian Follicle / Q000502:physiology; D018384:Oxidative Stress; D011374:Progesterone / Q000378:metabolism*; D011897:Random Allocation; D017382:Reactive Oxygen Species / Q000378:metabolism; D056566:Sirtuin 3 / Q000502:physiology*; D013482:Superoxide Dismutase / Q000378:metabolism; D000072105:Superoxide Dismutase-1
chemical_list: D000975:Antioxidants; D006063:Chorionic Gonadotropin; D017382:Reactive Oxygen Species; C000606290:SOD1 protein, human; D011374:Progesterone; D002374:Catalase; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; C466093:SIRT3 protein, human; D056566:Sirtuin 3