微囊藻毒素-LR对猪体外成熟卵母细胞氧化损伤与凋亡的影响

doi:10.11843/j.issn.0366-6964.2021.010

Abstract

Abstract: The aim of this study was to investigate the toxic damage effect of microcystin-LR (MC-LR) on porcine oocytes in vitro and its potential mechanism. The GV oocytes were collected from porcine ovaries obtained from a local slaughterhouse and randomly assigned into 4 groups, 0, 20, 40 and 60 μg·mL^-1 MC-LR were added into maturation medium in vitro, respectively. Then, the effects of MC-LR on the first polar body (PbI) extrusion, spindle structure, reactive oxygen species (ROS), membrane mucin V(Annexin V) and glutathione peroxidase (GSH-Px) activity were examined, respectively. The mRNA expression of oxidative stress and apoptosis related genes (SOD1, SOD2, CAT, GSH-Px, Bax and Bcl2) were also analyzed by RT-qPCR, and three replicates were performed for each experiment. The result showed that the PbI extrusion rate of porcine oocytes decreased in a concentration-dependent manner after MC-LR treatment. When the MC-LR concentration reached more than 40 μg·mL^-1, the oocyte maturation rate extremely significantly decreased(P<0.01), and the spindle structure abnormality rate also extremely significantly increased(P<0.001). Further studies showed that, after MC-LR treatment, ROS levels in oocytes were extremely significantly increased (P<0.01), while GSH-Px activity was extremely significantly decreased (P<0.01), accompanied by extremely significant down-regulation of antioxidant related genes SOD1, CAT and GSH-Px mRNA expressions(P<0.01). Apoptosis detection showed that MC-LR treatment could extremely significantly increase the early apoptosis rate of oocytes (P<0.01), and the ratio of Bax/Bcl2 was extremely significantly increased (P<0.001). The results indicate that MC-LR can induce abnormal spindle structure of porcine oocytes, lead to oxidative stress and apoptosis, and eventually result in a decline of porcine oocyte maturation ability.

Key words: microcystin-LR, porcine, oocyte, spindle, oxidative stress, apoptosis

CLC Number:

S828.3

SHI Fengyao, LI Wenhui, ZHAO Hongyu, WANG Yang, ZHUANG Ruixue, RUI Rong, JU Shiqiang. Effects of MC-LR on Oxidative Stress and Apoptosis of Porcine Oocytes in Vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(1): 98-106.

References 41

[1]	IBELINGS B W,CHORUS I.Accumulation of cyanobacterial toxins in freshwater "seafood" and its consequences for public health:a review[J].Environ Pollut,2007,150(1):177-192.
[2]	CHEN J,ZHANG D W,XIE P,et al.Simultaneous determination of microcystin contaminations in various vertebrates (fish,turtle, duck and water bird) from a large eutrophic Chinese lake,Lake Taihu,with toxic Microcystis blooms[J].Sci Total Environ, 2009,407(10):3317-3322.
[3]	HUISMAN J,CODD G A,PAERL H W,et al.Cyanobacterial blooms[J].Nat Rev Microbiol,2018,16(8):471-483.
[4]	BOUAÏCHA N,MILES C O,BEACH D G,et al.Structural diversity,characterization and toxicology of Microcystins[J].Toxins (Basel), 2019,11(12):714.
[5]	MEREL S,WALKER D,CHICANA R,et al.State of knowledge and concerns on cyanobacterial blooms and cyanotoxins[J]. Environ Int,2013,59:303-327.
[6]	REZAITABAR S,SARI A E,BAHRAMIFAR N,et al.Transfer,tissue distribution and bioaccumulation of microcystin LR in the phytoplanktivorous and carnivorous fish in Anzali wetland,with potential health risks to humans[J].Sci Total Environ,2017, 575:1130-1138.
[7]	MACHADO J,CAMPOS A,VASCONCELOS V,et al.Effects of microcystin-LR and cylindrospermopsin on plant-soil systems:a review of their relevance for agricultural plant quality and public health[J].Environ Res,2017,153:191-204.
[8]	GREER B,MENEELY J P,ELLIOTT C T.Uptake and accumulation of Microcystin-LR based on exposure through drinking water:an animal model assessing the human health risk[J].Sci Rep,2018,8(1):4913.
[9]	GUPTA N,PANT S C,VIJAYARAGHAVAN R,et al.Comparative toxicity evaluation of cyanobacterial cyclic peptide toxin microcystin variants (LR,RR,YR) in mice[J].Toxicology,2003,188(2-3):285-296.
[10]	PUDDICK J,PRINSEP M R,WOOD S A,et al.High levels of structural diversity observed in microcystins from Microcystis CAWBG11 and characterization of six new microcystin congeners[J].Mar Drugs,2014,12(11):5372-5395.
[11]	CHERNOFF N,HILL D,LANG J,et al.The comparative toxicity of 10 microcystin congeners administered orally to mice:clinical effects and organ toxicity[J].Toxins (Basel),2020,12(6):403.
[12]	CHEN J,XIE P.Tissue distributions and seasonal dynamics of the hepatotoxic microcystins-LR and -RR in two freshwater shrimps,Palaemon modestus and Macrobrachium nipponensis,from a large shallow,eutrophic lake of the subtropical China[J]. Toxicon, 2005,45(5):615-625.
[13]	WANG Q,XIE P,CHEN J,et al.Distribution of microcystins in various organs (heart,liver,intestine,gonad,brain,kidney and lung) of Wistar rat via intravenous injection[J].Toxicon,2008,52(6):721-727.
[14]	YI X P,XU S S,HUANG F Y,et al.Effects of chronic exposure to microcystin-LR on kidney in mice[J].Int J Environ Res Public Health,2019,16(24):5030.
[15]	LIN W,GUO H H,WANG L K,et al.Parental transfer of microcystin-LR-induced innate immune dysfunction of zebrafish:a cross-generational study[J].Environ Sci Technol,2020,54(2):1014-1023.
[16]	HOU J,LI L,XUE T,et al.Damage and recovery of the ovary in female zebrafish i.p.-injected with MC-LR[J].Aquat Toxicol,2014,155:110-118.
[17]	ZHAO Y Y,XIE L Q,YAN Y Y.Microcystin-LR impairs zebrafish reproduction by affecting oogenesis and endocrine system[J]. Chemosphere,2015,120:115-122.
[18]	CHEN L,CHEN J,ZHANG X Z,et al.A review of reproductive toxicity of microcystins[J].J Hazard Mater,2016,301:381-399.
[19]	LIU W J,ZHAN C H,ZHANG T Z,et al.Microcystin-LR influences the in vitro oocyte maturation of zebrafish by activating the MAPK pathway[J].Aquat Toxicol,2019,215:105261.
[20]	QIAO Q,DJEDIAT C,HUET H,et al.Subcellular localization of microcystin in the liver and the gonads of medaka fish acutely exposed to microcystin-LR[J].Toxicon,2019,159:14-21.
[21]	DING X S,LI X Y,DUAN H Y,et al.Toxic effects of Microcystis cell extracts on the reproductive system of male mice[J].Toxicon,2006,48(8):973-979.
[22]	HUANG H,LIU C R,FU X L,et al.Microcystin-LR induced apoptosis in rat sertoli cells via the mitochondrial caspase-dependent pathway:role of reactive oxygen species[J].Front Physiol,2016,7:397.
[23]	WU J,SHAO S J,ZHOU F,et al.Reproductive toxicity on female mice induced by microcystin-LR[J].Environ Toxicol Pharmacol,2014,37(1):1-6.
[24]	LIU H H,ZHANG X F,ZHANG S S,et al.Oxidative stress mediates microcystin-LR-induced endoplasmic reticulum stress and autophagy in KK-1 cells and C57BL/6 mice ovaries[J].Front Physiol,2018,9:1058.
[25]	WU J,YUAN M M,SONG Y F,et al.MC-LR exposure leads to subfertility of female mice and induces oxidative stress in granulosa cells[J].Toxins (Basel),2015,7(12):5212-5223.
[26]	BU Y Z,LI X Y,ZHANG B J,et al.Microcystins cause embryonic toxicity in mice[J].Toxicon,2006,48(8):966-972.
[27]	ZHAO S J,LI G Y,CHEN J.A proteomic analysis of prenatal transfer of microcystin-LR induced neurotoxicity in rat offspring[J].J Proteomics,2015,114:197-213.
[28]	ZHAO S J,ZHONG S Z,WANG F,et al.Microcystin-LR exposure decreased the fetal weight of mice by disturbance of placental development and ROS-mediated endoplasmic reticulum stress in the placenta[J].Environ Pollut,2020,256:113362.
[29]	ZHANG Z X,CHEN C C,MA L Y,et al.Plk1 is essential for proper chromosome segregation during meiosis I/meiosis II transition in pig oocytes[J].Reprod Biol Endocrinol,2017,15(1):69.
[30]	HUANG X,CHEN L,LIU W J,et al.Involvement of oxidative stress and cytoskeletal disruption in microcystin-induced apoptosis in CIK cells[J].Aquat Toxicol,2015,165:41-50.
[31]	MCLELLAN N L,MANDERVILLE R A.Toxic mechanisms of microcystins in mammals[J].Toxicol Res (Camb),2017,6(4):391-405.
[32]	FRANGEŽ R,ŽUŽEK M C,MRKUN J,et al.Microcystin-LR affects cytoskeleton and morphology of rabbit primary whole embryo cultured cells in vitro[J].Toxicon,2003,41(8):999-1005.
[33]	CHEN L,ZHANG X Z,ZHOU W S,et al.The interactive effects of cytoskeleton disruption and mitochondria dysfunction lead to reproductive toxicity induced by microcystin-LR[J].PLoS One,2013,8(1):e53949.
[34]	WENG D,LU Y,WEI Y N,et al.The role of ROS in microcystin-LR-induced hepatocyte apoptosis and liver injury in mice[J].Toxicology,2007,232(1-2):15-23.
[35]	MA J G,LI Y Y,DUAN H Y,et al.Chronic exposure of nanomolar MC-LR caused oxidative stress and inflammatory responses in HepG2 cells[J].Chemosphere,2018,192:305-317.
[36]	AGARWAL A,GUPTA S,SHARMA R.Oxidative stress and its implications in female infertility-a clinician's perspective[J]. Reprod Biomed Online,2005,11(5):641-650.
[37]	TRUONG T,GARDNER D K.Antioxidants improve IVF outcome and subsequent embryo development in the mouse[J].Hum Reprod,2017,32(12):2404-2413.
[38]	ZHONG S Z,LIU Y,WANG F,et al.Microcystin-LR induced oxidative stress,inflammation,and apoptosis in alveolar type II epithelial cells of ICR mice in vitro[J].Toxicon,2020,174:19-25.
[39]	FU W Y,CHEN J P,WANG X M,et al.Altered expression of p53,Bcl-2 and Bax induced by microcystin-LR in vivo and in vitro[J].Toxicon,2005,46(2):171-177.
[40]	LIU H H,ZHANG S S,LIU C R,et al.Resveratrol ameliorates microcystin-LR-induced testis germ cell apoptosis in rats via SIRT1 signaling pathway activation[J].Toxins,2018,10(6):235.
[41]	YUAN L,LIU H H,LIU X H,et al.Epigenetic modification of H3K4 and oxidative stress are involved in MC-LR-induced apoptosis in testicular cells of SD rats[J].Environ Toxicol,2020,35(2):277-291.

Effects of MC-LR on Oxidative Stress and Apoptosis of Porcine Oocytes in Vitro

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 41

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LI Wanjun, XU Jiehuan, HE Mengxian, KONG Yuting, ZHANG Defu, DAI Jianjun. Cytochalasin B Alleviates the Migration Disorder of Cortical Particle Caused by Vitrification in Porcine Oocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 1999-2010.
[2]	DONG Shucan, MAO Shuaixiang, WU Cuiying, LI Yaokun, SUN Baoli, GUO Yongqing, DENG Ming, LIU Dewu, LIU Guangbin. The Effect of the Androgen Receptor Inhibitor Enzalutamide on Proliferation and Apoptosis of Goat Ovarian Granulosa Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2022-2031.
[3]	HAN Yang, GUAN Shuaiyin, LI Zhen, ZHOU Saisai, YUAN Honggen, SONG Yunfeng. Prokaryotic Expression and Protein Activity of Porcine Circovirus Type 3 Rep [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2061-2071.
[4]	MA Rumeng, ZHAO Yuliang, MA Mingshuang, GUO Guihai, LIU Xinzi, LI Jiaxuan, CUI Wen, JIANG Yanping, SHAN Zhifu, ZHOU Han, WANG Li, QIAO Xinyuan, TANG Lijie, WANG Xiaona, LI Yijing. Comparative Study on the Immune Response Induced by the Different Porcine Receptor Bacteria with Expressing the Protective Antigen S1 of Porcine Epidemic Diarrhea Virus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2090-2099.
[5]	WANG Jing, ZHANG Shujuan, HU Xia, LIU Xiangyang, ZHANG Xingcui, SONG Zhenhui. CD44 Regulates Na⁺/H⁺ Exchanger 3 Activity by Influencing Porcine Epidemic Diarrhea Virus Replication [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2176-2185.
[6]	WANG Jiying, YIN Ruiru, XIE Xing, WANG Haiyan, LIU Hudong, HU Hui, XIONG Qiyan, FENG Zhixin, SHAO Guoqing, YU Yanfei. Effects of LDH in Mesomycoplasma (Mycoplasma) hyopneumoniae on Apoptosis of Porcine Bronchial Epithelial Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2195-2205.
[7]	LI Qiuyun, TIAN Xinyuan, LIAO Wensheng, ZHANG Huanrong, REN Yupeng, YANG Falong, ZHU Jiangjiang, XIANG Hua. Effects of SOCS2 on Proliferation, Cycle and Apoptosis of Turbinate Bone Cells in Goats [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2226-2240.
[8]	HU Zeqi, LI Runcheng, TAN Zuming, XIE Xiuyan, WANG Jiangping, QIN Lejuan, LI Rong, GE Meng. Establishment and Preliminary Application of PEDV, PoRVA and PDCoV TaqMan Triple RT-qPCR Assay [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2267-2272.
[9]	LAN Xinrui, ZHAO Baobao, ZHANG Bihan, LIN Xiaoyu, MA Huiming, WANG Yongsheng. Effects of β-sitosterol on Porcine Oocyte Maturation and Embryonic Development in Vitro [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1629-1637.
[10]	LIN Lili, ZHANG Mengdi, ZHU Linlin, MA Hailong, SUN Qi, HE Qigai, ZHANG Mengjia, LI Wentao. Establishment of Neutralizing Antibody Detection Method based on Recombinant Fluorescent Virus of Porcine Epidemic Diarrhea Virus GⅡb Strain [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1649-1660.
[11]	ZHANG Xinting, QIU Wenyue, PANG Xiaoyue, SU Yiman, YE Jiali, HUANG Jianjia, ZHOU Shuilian, TANG Zhaoxin, WANG Rongmei, SU Rongsheng. Effect of Asiatic Acid Alleviating Myocardial Injury Caused by Lipopolysaccharide through Inhibiting Oxidative Stress and Ferroptosis in Broilers [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1787-1799.
[12]	LI Yujun, HE Honghong, YANG Lixue, YANG Xiaogeng, LI Jian, ZHANG Huizhu. Advances in Regulation of Mammalian Embryonic Development by Mitochondrial Autophagy [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 905-912.
[13]	JIANG Lijun, ZONG Yunhe, LI Yunlei, CHEN Jilan, GENG Zhaoyu, SUN Yanyan, JIN Sihua. Research Progress of Antioxidant Application in Poultry Semen Storage [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 913-923.
[14]	HU Qiaoyan, ZHAI Xiangqin, LI Yidan, HAN Jiale, LEI Chuzhao, DANG Ruihua. Effects of bta-miR-101 on Proliferation, Apoptosis and Secretion of Bovine Testicular Sertoli Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1040-1051.
[15]	JING Yang, WANG Yumiao, LI Yang, CHANG Hui, MA Zhiqian, LI Zhiwei, XIAO Shuqi. Construction of MARC-145^ORF6 Cell Line Stably Overexpressing PRRSV M Protein and Its Effect on PRRSV Proliferation [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1159-1169.