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丙酮酸和乳酸代谢对猪卵母细胞体外核成熟的作用

Effects of pyruvate and lactate metabolism on in vitro nuclear maturation of porcine oocytes
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摘要 关于卵母细胞成熟过程中糖代谢的研究主要集中在糖酵解和PPP途径上,而对丙酮酸和乳酸代谢的报道较少。本研究据此探讨猪卵母细胞体外核成熟过程中丙酮酸和乳酸的代谢途径及对卵母细胞核成熟的影响。以不含能量物质的NCSU-23为基础培养基,添加不同浓度的丙酮酸钠或乳酸钠及单羧酸转运蛋白(MCT)抑制剂4-CIN、线粒体呼吸链抑制剂鱼藤酮或乳酸脱氢酶抑制剂草氨酸钠,培养猪卵丘-卵母细胞复合体(COCs)或裸卵(DOs),观察核成熟率。结果表明,添加15mmol/L丙酮酸钠时,COCs和DOs核成熟率最高,分别为(69.5±3.1)%和(54.7±2.6)%;同时添加4-CIN或鱼藤酮显著(P<0.05)降低COCs和DOs核成熟率;添加3mmol/L乳酸钠时COCs和添加10mmol/L乳酸钠时DOs的核成熟率达到最高,分别为(72.1±1.7)%和(41.1±2.3)%;同时添加草氨酸钠显著(P<0.05)降低COCs和DOs的核成熟率。以上结果说明:(1)猪COCs和DOs均能代谢丙酮酸和乳酸支持其核成熟;(2)丙酮酸通过MCT跨膜转运进入线粒体并经线粒体呼吸链代谢支持卵母细胞核成熟;(3)乳酸通过LDH催化生成丙酮酸和NADH来维持卵母细胞核成熟;(4)猪COCs比DOs对丙酮酸和乳酸的利用能力强,说明卵丘细胞对卵母细胞成熟至关重要。 While there are many studies on the effects of glycolysis and PPP on oocyte maturation,reports are limited on pyruvate and lactate metabolism during oocyte maturation.Thus,this study has explored the pathways and effects of pyruvate and lactate metabolism on in vitro nuclear maturation of porcine oocytes.Porcine cumulus-oocyte complexes(COCs)and denuded oocytes(DOs)were matured in NCSU-23 medium supplemented with different concentrations of pyruvate sodium or lactate sodium with or without monocarboxylate transporter(MCT)inhibitorα-cyano-4-hydroxy cinnamate(4-CIN),mitochondrial respiratory chain inhibitor rotenone,or lactate dehydrogenase(LDH)inhibitor sodium oxamate before examination for nuclear maturation.The results showed that nuclear maturation rates of both COCs(69.5±3.1)%and DOs(54.7±2.6)%reached the highest when 15 mmol/L sodium pyruvate was added,and a simultaneous treatment with 4-CIN or rotenone significantly(P<0.05)decreased percentages of MII oocytes.Nuclear maturation rates of COCs(72.1 ± 1.7)% and DOs(41.1 ± 2.3)% reached the highest when3 mmol/L and 10 mmol/L sodium lactate were added,respectively,and a simultaneous treatment with sodium oxamate significantly(P<0.05)decreased the percentage of MII oocytes.Taken together,the results suggested that(i)both pig COCs and DOs could metabolize pyruvate and lactate to support nuclear maturation;(ii)pyruvate was transferred by MCT and utilized through mitochondrial electron transport to sustain oocyte nuclear maturation;(iii)lactate was oxidized to pyruvate by LDH to maintain oocyte maturation;and(iv)porcine COCs showed a higher capacity than DOs to utilize pyruvate and lactate,emphasizing the importance of cumulus cells in supporting oocyte maturation.
作者 温晶 王国梁 巩帅 解洪立 谭景和 WEN Jing;WANG Guo-liang;GONG Shuai;XIE Hong-li;TAN Jing-he(College of Life Science ,Northeast Agricultural University,Harbin 150030,China;College of Animal Science and Technology,Shandong Agricultural University,Tailan ,Shandong 271018,China)
出处 《中国兽医学报》 CSCD 北大核心 2018年第12期2404-2409,共6页 Chinese Journal of Veterinary Science
基金 国家重点研发计划资助项目(2017YFC1001601,2017YFC1001602) 国家重大基础研究计划资助项目(2014CB138503) 国家自然科学基金资助项目(31772599,31702114) 东北农业大学研究基金资助项目(106518001) 山东省双一流建设计划资助项目(SYL2017YSTD12).
关键词 丙酮酸 乳酸 核成熟 猪卵母细胞 pyruvate lactate nuclear maturation porcine oocytes
作者简介 温晶(1993-),女,硕士;通讯作者:谭景和,E-mail:tanjh@sdau.edu.cn
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