褪黑素保护卵母细胞应对热应激损伤的研究
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摘要
目的探讨褪黑素对热激条件下小鼠卵母细胞体外成熟的潜在影响。方法采用小鼠卵母细胞成熟的热激模型,将卵母细胞随机分成3组,分别是对照组、热激组和热激+褪黑素组,测定1×10-9 mol/L褪黑素对热激后卵母细胞质量的改善作用。结果与对照组相比,热激组小鼠卵母细胞成熟率显著下降[(33.00±0.07)%vs.(85.00±0.03)%,P<0.01],纺锤体组装异常,早期凋亡比率显著增加[(59.7±4.5)%vs.(22.0±3.5)%,P<0.01];热激+褪黑素组与热激组相比,卵母细胞成熟率显著上升[(70.00±0.05)%vs.(33.00±0.07)%,P<0.01],纺锤体异常比率显著下降,发生早期凋亡的卵母细胞比率亦显著下降[(37.3±6.1)%vs.(59.7±4.5)%,P<0.01]。热激+褪黑素组热休克蛋白70的表达水平与对照组和热激组相比显著上调(P<0.01)。结论褪黑素能通过调节热激后热休克蛋白70过表达来改善热激对卵母细胞造成的成熟率下降、纺锤体组装异常和早期凋亡比率的增加。
Objective:To explore the potential effect of melatonin on the in vitro maturation of mouse oocytes under heat shock condition.Methods:This study used a heat shock model of mouse oocyte maturation.The oocytes were randomly divided into three groups:control group,heat shock group and heat shock plus melatonin group.The effect of 1×10-9 mol/L melatonin on the quality of oocytes after heat shock was measured.Results:The maturation rate of mouse oocytes was significantly decreased under heat shock condition compared the control group[(33.00±0.07)% vs.(85.00±0.03)%,P<0.01].The assembly of spindle was abnormal and the early apoptosis rate was significantly increased[(59.7±4.5)% vs.(22.0±3.5)%,P<0.01].After adding melatonin,the oocytes maturation rate was significantly increased compared with the heat shock group[(70.00±0.05)% vs.(33.00±0.07)%,P<0.01],and the abnormal rate of spindle assembly and the early apoptosis rate were significantly decreased[(37.3±6.1)%vs.(59.7±4.5)%,P<0.01].The expression level of heat shock protein 70 was significantly up-regulated in the heat shock plus melatonin group compared with the other two groups(P<0.01).Conclusions:Melatonin can improve the decline of maturation rate on oocytes,the abnormality of spindle assembly,and the increase of early apoptosis rate caused by heat shock by regulating the expression of heat shock protein 70.
Objective:To explore the potential effect of melatonin on the in vitro maturation of mouse oocytes under heat shock condition.Methods:This study used a heat shock model of mouse oocyte maturation.The oocytes were randomly divided into three groups:control group,heat shock group and heat shock plus melatonin group.The effect of 1×10-9 mol/L melatonin on the quality of oocytes after heat shock was measured.Results:The maturation rate of mouse oocytes was significantly decreased under heat shock condition compared the control group[(33.00±0.07)% vs.(85.00±0.03)%,P<0.01].The assembly of spindle was abnormal and the early apoptosis rate was significantly increased[(59.7±4.5)% vs.(22.0±3.5)%,P<0.01].After adding melatonin,the oocytes maturation rate was significantly increased compared with the heat shock group[(70.00±0.05)% vs.(33.00±0.07)%,P<0.01],and the abnormal rate of spindle assembly and the early apoptosis rate were significantly decreased[(37.3±6.1)%vs.(59.7±4.5)%,P<0.01].The expression level of heat shock protein 70 was significantly up-regulated in the heat shock plus melatonin group compared with the other two groups(P<0.01).Conclusions:Melatonin can improve the decline of maturation rate on oocytes,the abnormality of spindle assembly,and the increase of early apoptosis rate caused by heat shock by regulating the expression of heat shock protein 70.
引文
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[14]Zeng F,Tee C,Liu M,et al.The p53/HSP70inhibitor,2-phenylethynesulfonamide,causes oxidative stress,unfolded protein response and apoptosis in rainbow trout cells[J].Aquat Toxicol,2014,146:45-51.
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[16]佘美华,蒋文艳,张瑶,等.褪黑素通过抑制ROS改善胰岛素抵抗HepG2细胞糖代谢[J].中南医学科学杂志,2014,42:433-435.
[17]Liu S,Dai X,Cai L,et al.Effect of Hsp27on early embryonic development in the mouse[J/OL].Reprod Biomed Online,2013,26:491-499.
[18]樊莹,唐敏,傅扬,等.褪黑激素在视网膜色素上皮细胞氧化应激中对线粒体DNA保护作用的研究[J].眼科新进展,2006,26:911-915.
[2]Nteeba J,Sanz-Fernandez MV,Rhoads RP,et al.Heat Stress alters ovarian insulin-mediated phosphatidylinositol-3kinase and steroidogenic signaling in gilt ovaries[J].Biol Reprod,2015,92:148.
[3]Tremblay P,Gori A,Maguer JF,et al.Heterotrophy promotes the re-establishment of photosynthate translocation in a symbiotic coral after heat stress[J].Sci Rep,2016,6:38112.
[4]Pavani KC,Baron E,Correia P,et al.Gene expression,oocyte nuclear maturation and developmental competence of bovine oocytes and embryos produced after in vivo and in vitro heat shock[J].Zygote,2016,24:748-759.
[5]Paula FF,Hansen PJ.Apoptosis is an adaptive response in bovine preimplantation embryos that facilitates survival after heat shock[J].Biochem Biophys Res Commun,2002,295:37-42.
[6]Gharibzadeh Z,Riasi A,Ostadhosseini S,et al.Effects of heat shock during the early stage of oocyte maturation on the meiotic progression,subsequent embryonic development and gene expression in ovine[J].Zygote,2015,23:573-582.
[7]Wang F,Tian X,Zhang L,et al.Beneficial effect of resveratrol on bovine oocyte maturation and subsequent embryonic development after in vitro fertilization[J].Fertil Steril,2014,101:577-586.
[8]王伟周,任立坤,沈玉良,等.褪黑素对老化卵母细胞体外发育能力的改善效果评价[J].生殖医学杂志,2017,26:470-476.
[9]Lan M,Han J,Pan MH,et al.Melatonin protects against defects induced by deoxynivalenol during mouse oocyte maturation[J].J Pineal Res,2018,65:e12477.
[10]Sirotkin AV.Effect of two types of stress(heat shock/high temperature and malnutrition/serum deprivation)on porcine ovarian cell functions and their response to hormones[J].Exp Biol 2010,213:377-382.
[11]Isom SC,Prather RS,Rucker EB.Heat stress-induced apoptosis in porcine in vitro fertilized and parthenogenetic preimplantationstage embryos[J].Mol Reprod Dev 2007,74:574-581.
[12]Hosseini SM,Hajian M,Nasr-Esfahani MH.Effects of heat shock during the early stage of oocyte maturation on the meiotic progression,subsequent embryonic development and gene expression in ovine[J].Zygote,2015,23:573-582.
[13]王杨杨,李小静.HSP70小干扰RNA对人瘢痕成纤维细胞胶原表达的影响[J].山东医药,2018,58:17-20.
[14]Zeng F,Tee C,Liu M,et al.The p53/HSP70inhibitor,2-phenylethynesulfonamide,causes oxidative stress,unfolded protein response and apoptosis in rainbow trout cells[J].Aquat Toxicol,2014,146:45-51.
[15]Wiech M,Olszewski MB,Tracz-Gaszewska Z,et al.Molecular mechanism of mutant p53stabilization:the role of HSP70and MDM2[J].Plos One,2012,7:e51426.
[16]佘美华,蒋文艳,张瑶,等.褪黑素通过抑制ROS改善胰岛素抵抗HepG2细胞糖代谢[J].中南医学科学杂志,2014,42:433-435.
[17]Liu S,Dai X,Cai L,et al.Effect of Hsp27on early embryonic development in the mouse[J/OL].Reprod Biomed Online,2013,26:491-499.
[18]樊莹,唐敏,傅扬,等.褪黑激素在视网膜色素上皮细胞氧化应激中对线粒体DNA保护作用的研究[J].眼科新进展,2006,26:911-915.