[1]李樊,李隐侠,舒嘉傲,等.热应激通过钙信号调控湖羊卵巢颗粒细胞凋亡和雌激素合成[J].江苏农业学报,2022,38(05):1278-1285.[doi:doi:10.3969/j.issn.1000-4440.2022.05.015]
　LI Fan,LI Yin-xia,SHU Jia-ao,et al.Regulation of heat stress on ovarian granulosa cell apoptosis and estrogen synthesis in Hu sheep through calcium signal[J].,2022,38(05):1278-1285.[doi:doi:10.3969/j.issn.1000-4440.2022.05.015]
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热应激通过钙信号调控湖羊卵巢颗粒细胞凋亡和雌激素合成()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
38
期数:
2022年05期
页码:
1278-1285
栏目:
畜牧兽医·水产养殖
出版日期:
2022-10-31

文章信息/Info

Title:
Regulation of heat stress on ovarian granulosa cell apoptosis and estrogen synthesis in Hu sheep through calcium signal
作者:
李樊1; 2; 3; 李隐侠1; 2; 3; 舒嘉傲1; 2; 3; 孟春花1; 2; 3; 张俊1; 2; 3; 钱勇1; 2; 3; 曹少先1; 2; 3
（1.江苏省农业科学院畜牧研究所，江苏南京210014；2.江苏省农业种质资源保护与利用平台，江苏南京210014；3.农业农村部种养结合重点实验室，江苏南京210014）
Author(s):
LI Fan1; 2; 3; LI Yin-xia1; 2; 3; SHU Jia-ao1; 2; 3; MENG Chun-hua1; 2; 3; ZHANG Jun1; 2; 3; QIAN Yong1; 2; 3; CAO Shao-xian1; 2; 3
(1.Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;2.Jiangsu Agricultural Germplasm Resources Protection and Utilization Platform, Nanjing 210014, China;3.Key Laboratory of Combination of Planting and Breeding, Ministry of Agriculture and Rural Affairs, Nanjing 210014, China)
关键词:
热应激; Ca2+; 卵巢颗粒细胞; 细胞凋亡; 雌二醇
Keywords:
heat stress; Ca2+; ovarian granulosa cells; cell apoptosis; estradiol
分类号:
S826
DOI:
doi:10.3969/j.issn.1000-4440.2022.05.015
文献标志码:
A
摘要:
热应激严重影响动物的繁殖性能，但热应激是如何通过调控绵羊卵巢颗粒细胞功能进而调控绵羊繁殖的相关研究较少。本研究以湖羊卵巢颗粒细胞为研究对象，利用Real-time PCR、流式细胞技术、Western blot、ELISA等方法探讨热应激对湖羊卵巢颗粒细胞钙信号通路、细胞凋亡和雌激素合成的影响。结果表明，热应激后，湖羊卵巢颗粒细胞中钙离子浓度显著上升（P＜0.05），钙离子信号通路中相关基因CACNA1B、CACNA1C、RYR1、CAMK2表达量显著升高，线粒体分裂蛋白FIS1表达量显著上升，促凋亡蛋白家族代表基因BAX和抗凋亡蛋白代表基因BCL2 mRNA表达量比值（BAX/BCL2）显著上升，细胞凋亡率增加；雌二醇合成关键酶CYP19A1表达量显著下调，雌二醇含量显著下降（P＜0.05）。热应激卵巢颗粒细胞中加入钙离子螯合剂（BAPTA-AM）明显降低钙离子浓度，同时减少细胞凋亡，缓解热应激造成的卵巢颗粒细胞雌二醇含量下调。综上表明，热应激打开了湖羊卵巢颗粒细胞膜电压门控钙离子通道和内质网膜上钙离子释放通道，导致细胞质中钙离子浓度上升，促进卵巢颗粒细胞凋亡，引起雌二醇含量下调。
Abstract:
Heat stress seriously affects the reproductive performance of animals, but there are few studies on the effects of heat stress on sheep reproduction by regulating the function of sheep ovarian granulosa cells. In this study, Hu sheep ovarian granulosa cells were used as the research objects, and real-time PCR, flow cytometry, Western blot, ELISA and other methods were used to investigate the effects of heat stress on calcium signaling pathway, apoptosis and estrogen synthesis in Hu sheep ovarian granulosa cells. The results showed that after heat stress, the concentration of Ca2+ in Hu sheep ovarian granulosa cells increased significantly (P＜0.05), the expression levels of calcium signaling pathway-related genes CACNA1B, CACNA1C, RYR1 and CAMK2 and mitochondrial fission protein FIS1 were significantly increased, the mRNA expression ratio of pro-apoptotic protein family representative gene BAX and anti-apoptotic protein family representative gene BCL2 (BAX/BCL2) increased significantly, and the apoptosis rate increased. The expression of CYP19A1, the key enzyme in estradiol synthesis, was down-regulated, and the content of estradiol was significantly decreased (P＜0.05). The addition of calcium chelator BAPTA-AM to heat-stressed ovarian granulosa cells significantly reduced Ca2+ concentration, decreased cell apoptosis, and alleviated the down-regulation of estradiol content in ovarian granulosa cells caused by heat stress. In conclusion, heat stress opens the voltage-gated calcium ion channel on the cell membrane of Hu sheep ovarian granulosa cells and the calcium ion release channel on the endoplasmic reticulum membrane, which leads to the increase of calcium ion concentration in the cytoplasm, promotes the apoptosis of ovarian granulosa cells, and causes the down-regulation of estradiol content.

参考文献/References:

[1]NICOLS-LPEZ P, MACAS-CRUZ U, MELLADO M, et al. Growth performance and changes in physiological, metabolic and hematological parameters due to outdoor heat stress in hair breed male lambs finished in feedlot [J]. International Journal Biometeorology, 2021, 65: 1451-1459.
[2]MACAS-CRUZ U, SAAVEDRA O R, CORREA-CALDERN A, et al. Feedlot growth, carcass characteristics and meat quality of hair breed male lambs exposed to seasonal heat stress (winter vs. summer) in an arid climate [J]. Meat Science, 2020, 169:108202.
[3]MEZA-HERRERA C A, VICENTE-PEREZ A, OSORIO-MARIN Y, et al. Heat stress, divergent nutrition level, and late pregnancy in hair sheep: effects upon cotyledon development and litter weight at birth [J]. Tropical Animal Health and Production, 2015, 47(5): 819-824.
[4]KLEEMANN D O, WALKER S K. Fertility in South Australian commercial Merino flocks: relationships between reproductive traits and environmental cues [J]. Theriogenology, 2005, 63(9): 2416-2433.
[5]SLUSARSKI D C, PELEGRI F. Calcium signaling in vertebrate embryonic patterning and morphogenesis[J]. Developmental Biology, 2007, 307(1):1-13.
[6]BERRIDGE M J, BOOTMAN M D, RODERICK H L. Calcium signalling: dynamics, homeostasis and remodelling [J]. Nature Reviews Molecular Cell Biology, 2003, 4:517-529.
[7]RUSSA A D, MAESAWA C, SATOH Y. Spontaneous [Ca2+]i oscillations in G1/S phase-synchronized cells [J]. Journal of Electron Microscopy, 2009, 58（5）:321-329.
[8]PATEL R, HOLT M, PHILIPOVA R, et al. Calcium/calmodulin-dependent phosphorylation and activation of human Cdc25-C at the G2/M phase transition in HeLa cells [J]. Journal of Biological Chemistry, 1999, 274(12):7958-7968.
[9]HEIM A, TISCHER T, MAYER T U. Calcineurin promotes APC/C activation at meiotic exit by acting on both XErp1 and Cdc20 [J]. EMBO Reports, 2018, 19(12): e46433.
[10]MARIN M. Calcium signaling in Xenopus oocyte [J]. Advances in Experimental Medicine and Biology, 2012,740:1073-1094.
[11]CHRISTOU M, FRASER T W K, BERG V, et al. Calcium signaling as a possible mechanism behind increased locomotor response in zebrafish larvae exposed to a human relevant persistent organic pollutant mixture or PFOS [J]. Environmental Research, 2020, 187:109702.
[12]MIAO Y L, WILLIAMS C J. Calcium signaling in mammalian egg activation and embryo development: influence of subcellular localization[J]. Molecular Reproduction and Development, 2012, 79(11):742-756.
[13]LI Y K, KONG L X, DENG M, et al. Heat stress-responsive transcriptome analysis in the liver tissue of Hu sheep [J]. Genes, 2019, 10(5): 395.
[14]KUMAR D, DE K, SHEKHAWAT I, et al. Combined effect of heat and nutritional stress on superovulation of Malpura ewes in a semi-arid region[J]. Journal of Thermal Biology, 2019, 80:158-163.
[15]LI L, TAN H P, GU Z T, et al. Heat stress induces apoptosis through a Ca2+-mediated mitochondrial apoptotic pathway in human umbilical vein endothelial cells[J]. PLoS One, 2014, 9(12): e111083.
[16]LI Y X, FENG X P, WANG H L, et al. Transcriptome analysis reveals corresponding genes and key pathways involved in heat stress in Hu sheep [J]. Cell Stress and Chaperones, 2019, 24(6):1045-1054.
[17]LI F, LONG Y, XIE J, et al. Generation of GCaMP6s-expressing zebrafish to monitor spatiotemporal dynamics of calcium signaling elicited by heat stress [J]. International Journal Molecular Sciences, 2021, 22(11):5551.
[18]李双君，潘君，崔玉红. 内皮细胞电压门控钙离子通道及其功能研究进展[J].生物化学与生物物理进展，2022，49（6）：1061-1074.
[19]LAVER D R. Ca2+ stores regulate ryanodine receptor Ca2+ release channels via luminal and cytosolic Ca2+ sites [J]. Biophysical Journal, 2007, 92(10):3541-3555.
[20]HOEFLICH K P, IKURA M. Calmodulin in action: diversity in target recognition and activation mechanisms [J]. Cell, 2002,108:739-742.
[21]李文俊,李强,钟良明,等. 基于线粒体DNA控制区序列的珠江和长江水系光倒刺鲃群体遗传变异分析[J].南方农业学报,2021,52(11):3121-3129.
[22]谢启明,柯瑞林,刘帆,等. 基于线粒体D-loop区分析安徽省5个翘嘴鳜养殖群体的遗传多样性[J].江苏农业科学,2021,49(9):143-147.
[23]邹辉,韦玲静,黄杰,等. 基于线粒体COⅠ基因和D-Loop区序列的7个鲤群体遗传差异分析[J].南方农业学报,2020,51(5):1209-1216.
[24]陶元，彭凯歌，但国蓉，等.线粒体融合/分裂对线粒体的质量控制作用 [J].现代生物医学进展, 2015, 15（23）：4588-4590.
[25]SLIMEN I B, NAJAR T, GHRAM A, et al. Reactive oxygen species, heat stress and oxidative-induced mitochondrial damage. A review [J]. International Journal Hyperthermia, 2014, 30(7):513-523.
[26]马汉宁，姬艳燕，陈伟，等. 甘草次酸通过抑制Caspase 3/Bax/Bcl-2凋亡信号通路保护心脏骤停心肺复苏大鼠心脏功能[J]. 中药药理与临床，2019, 35(4): 28-33.
[27]KARIMI A S， TAFVIZI F， TAJABADI E M. Heat-killed probiotic bacteria induce apoptosis of HT-29 human colon adenocarcinoma cell line via the regulation of Bax/Bcl2 and caspases pathway[J]. Human & Experimental Toxicology，2019，38 (9): 1069-1081.
[28]ROSENFELD C S, WAGNER J S, ROBERTS R M, et al. Intraovarian actions of oestrogen[J]. Reproduction, 2001, 122(2): 215-226.

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备注/Memo

备注/Memo:
收稿日期：2021-12-18基金项目：江苏省农业科技自主创新基金项目［CX(20)3013］；国家自然科学基金项目(31902155)作者简介：李樊（1997-），女，湖北孝感人，硕士研究生，主要从事动物分子生物学研究。（E-mail）lf15205159110@163.com通讯作者：曹少先，（E-mail）sxcao@jaas.ac.cn
更新日期/Last Update: 2022-11-07