title: Coagulansin-A has beneficial effects on the development of bovine embryos in vitro via HSP70 induction.
abstract: Coagulansin-A (withanolide) is the steroidal lactone obtained from Withania coagulans which belong to Solanaceae family. The present study investigated the effects of coagulansin-A on bovine oocyte maturation and embryo development in vitro. All these oocytes were aspirated from the ovaries obtained from Korean Hanwoo cows at a local abattoir. To determine whether coagulansin-A has beneficial effects on bovine oocyte maturation in vitro, 355 oocytes per group (control and treated) in seven replicates were subjected with different concentrations (1, 2.5, 5, 7.5 and 10 μM) of coagulansin-A. The coagulansin-A was added in the in vitro maturation (IVM) media followed by in vitro fertilization (IVF) and then in vitro culture (IVC). Only treatment with 5 μM coagulansin-A remarkably (P<0.05) improved embryos development (Day 8 blastocyst) having 27.30 and 40.01% for control and coagulansin-A treated groups respectively. Treatment with 5 μM coagulansin-A significantly induced activation of heat shock protein 70 (HSP70) (P<0.05). Immunofluorescence analysis revealed that 5 μM coagulansin-A treatment also significantly inhibited oxidative stress and inflammation during bovine embryo development in vitro by decreasing 8-oxoguanosine (8-OxoG) (P<0.05) and nuclear factor-κB (NF-κB) (P<0.05). The expressions of HSP70 and NF-κB were also conformed through real-time PCR (RT-PCR). Additionally, the terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assay confirmed that coagulansin-A treatment significantly improved the embryo quality and reduced bovine embryo DNA damage (P<0.05). The present study provides new information regarding the mechanisms by which coagulansin-A promotes bovine embryo development in vitro.
authors.affiliation: Department of Animal Science, Division of Applied Life Science (BK21 Plus), Gyeongsang National University, Jinju 660-701, Gyeongnam Province, Republic of Korea.
authors.forename: Imran
authors.initials: I
authors.lastname: Khan
authors.affiliation: Department of Animal Science, Division of Applied Life Science (BK21 Plus), Gyeongsang National University, Jinju 660-701, Gyeongnam Province, Republic of Korea.
authors.forename: Kyeong-Lim
authors.initials: KL
authors.lastname: Lee
authors.affiliation: Department of Animal Science, Division of Applied Life Science (BK21 Plus), Gyeongsang National University, Jinju 660-701, Gyeongnam Province, Republic of Korea.
authors.forename: Md
authors.initials: M
authors.lastname: Fakruzzaman
authors.affiliation: Department of Animal Science, Division of Applied Life Science (BK21 Plus), Gyeongsang National University, Jinju 660-701, Gyeongnam Province, Republic of Korea.
authors.forename: Seok-Hwan
authors.initials: SH
authors.lastname: Song
authors.affiliation: Department of Pharmacy, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad 45320, Pakistan.
authors.lastname: Ihsan-ul-Haq
authors.affiliation: Department of Biochemistry, Quaid-i-Azam University, Islamabad 45320, Pakistan.
authors.forename: Bushra
authors.initials: B
authors.lastname: Mirza
authors.affiliation: Jilin Co-Innovation Center of Beef Cattle Science and Industry Technology, Yanbian University, Yanji, Jilin 133002, P.R. China.
authors.forename: Chang Guo
authors.initials: CG
authors.lastname: Yan
authors.affiliation: Department of Animal Science, Division of Applied Life Science (BK21 Plus), Gyeongsang National University, Jinju 660-701, Gyeongnam Province, Republic of Korea Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Gyeongnam Province, Republic of Korea ikong7900@gmail.com.
authors.forename: Il-Keun
authors.initials: IK
authors.lastname: Kong
mesh_terms: D000818:Animals; D002417:Cattle; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D047108:Embryonic Development / Q000187:drug effects*; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis*; D054358:Withanolides / Q000494:pharmacology*
chemical_list: C433551:3-rhamnopyranosyl(1-4)-glucopyranosyl-12-diacetoxy-20-hydroxywitha-5,24-dienolide; D018840:HSP70 Heat-Shock Proteins; D054358:Withanolides
keywords: DNA damage; bovine embryo; coagulansin-A; heat shock protein 70 (HSP70); nuclear factor-κB (NF-κB)
references.citation: Biol Reprod. 2000 Jun;62(6):1526-35
references.pmid: 10819752
references.citation: Biochem Pharmacol. 2014 Oct 15;91(4):501-9
references.pmid: 25159986
references.citation: Nature. 2000 Nov 9;408(6809):239-47
references.pmid: 11089981
references.citation: Biol Reprod. 2002 Mar;66(3):589-95
references.pmid: 11870062
references.citation: Biol Reprod. 2003 Jan;68(1):236-43
references.pmid: 12493719
references.citation: Reprod Domest Anim. 2003 Aug;38(4):259-67
references.pmid: 12887565
references.citation: Theriogenology. 2004 Oct 1;62(7):1186-97
references.pmid: 15325546
references.citation: Hum Mol Genet. 2004 Oct 1;13(19):2263-78
references.pmid: 15317747
references.citation: Adv Exp Med Biol. 1987;225:287-304
references.pmid: 3331062
references.citation: Annu Rev Genet. 1988;22:631-77
references.pmid: 2853609
references.citation: Indian J Exp Biol. 1988 Nov;26(11):877-82
references.pmid: 3248848
references.citation: Chem Biol Interact. 1989;72(1-2):1-56
references.pmid: 2684438
references.citation: Mol Reprod Dev. 1991 Jan;28(1):1-8
references.pmid: 1994975
references.citation: Annu Rev Biochem. 1991;60:321-47
references.pmid: 1679318
references.citation: Biol Reprod. 1993 Sep;49(3):459-62
references.pmid: 8399836
references.citation: Eur J Biochem. 1994 Jan 15;219(1-2):11-23
references.pmid: 8306977
references.citation: J Reprod Fertil. 1993 Nov;99(2):673-9
references.pmid: 8107053
references.citation: Annu Rev Genet. 1993;27:437-96
references.pmid: 8122909
references.citation: Experientia. 1994 Nov 30;50(11-12):1031-8
references.pmid: 7988662
references.citation: Biol Reprod. 1995 Oct;53(4):786-90
references.pmid: 8547471
references.citation: J Anim Sci. 1997 Feb;75(2):483-9
references.pmid: 9051472
references.citation: Mol Reprod Dev. 1997 Sep;48(1):25-33
references.pmid: 9266758
references.citation: Rev Reprod. 1997 Jan;2(1):48-54
references.pmid: 9414465
references.citation: Biol Reprod. 1999 Jun;60(6):1536-41
references.pmid: 10330116
references.citation: Nucleic Acids Res. 2006;34(11):3279-87
references.pmid: 16807318
references.citation: Biochem Pharmacol. 2006 Nov 30;72(11):1493-505
references.pmid: 16723122
references.citation: Reproduction. 2007 Jan;133(1):95-106
references.pmid: 17244736
references.citation: Reproduction. 2007 May;133(5):947-54
references.pmid: 17616724
references.citation: Theriogenology. 2007 Sep 1;68 Suppl 1:S131-7
references.pmid: 17467789
references.citation: Cell. 2008 Feb 8;132(3):344-62
references.pmid: 18267068
references.citation: Hum Reprod Update. 2008 Mar-Apr;14(2):131-42
references.pmid: 18239135
references.citation: Clin Sci (Lond). 2009 Mar;116(6):451-65
references.pmid: 19200055
references.citation: J Pharm Pharmacol. 2010 Feb;62(2):153-60
references.pmid: 20487193
references.citation: J Anim Sci. 2011 Sep;89(9):2759-67
references.pmid: 21478451
references.citation: PLoS One. 2012;7(5):e37764
references.pmid: 22701533
references.citation: J Nat Prod. 2013 Jan 25;76(1):22-8
references.pmid: 23316950
references.citation: Zygote. 2013 Aug;21(3):250-5
references.pmid: 21791164
references.citation: Dev Dyn. 2000 Nov;219(3):333-40
references.pmid: 11066090