SRIF-14对绵羊子宫内膜上皮细胞增殖和凋亡的影响

doi:10.11843/j.issn.0366-6964.2023.08.018

摘要/Abstract

摘要： 旨在初步探索生长抑素（somatotropin release inhibitory factor，SRIF）在绵羊子宫内膜上皮细胞中的作用。本研究中，绵羊子宫组织样品来自新疆石河子市牛羊定点屠宰场，利用免疫组织化学染色方法明确SRIF在绵羊子宫内膜中的表达情况；使用雌二醇（estradiol，E₂）、孕酮（progesterone，P₄）和干扰素-tau （interferon-tau，IFNT）模拟绵羊子宫内膜中胚胎植入时的激素变化水平，RT-qPCR和ELISA检测促孕体延长因子和前列腺素（prostaglandins，PGs）的表达，并使用Western blot方法检测SRIF的表达水平；在绵羊子宫内膜上皮细胞中添加SRIF-14，使用CCK-8试剂盒和RT-qPCR法检测细胞增殖能力；Western blot方法检测Caspase 9、Bax、Bcl-2和Cyt-c的蛋白表达水平；ELISA试剂盒检测细胞分泌PGE₂和PGF_2α的水平。免疫组织化学染色结果显示，SRIF在绵羊子宫内膜组织中普遍表达，且细胞染色结果证实SRIF在绵羊子宫内膜上皮细胞中表达。激素联合处理绵羊子宫内膜上皮细胞的结果显示，促孕体伸长因子ISG15、CXCL10、HOXA10和RSAD2 mRNA表达水平显著升高，PGE₂分泌水平提高、PGF_2α分泌水平降低，SRIF的蛋白表达水平在激素处理后显著上升。在绵羊子宫内膜上皮细胞中外源性添加SRIF-14的结果表明，SRIF-14抑制绵羊子宫内膜上皮细胞的增殖能力，CCNA mRNA表达水平显著下降；Bax、Csapase 9和Cyt-c蛋白表达水平显著升高，Bcl-2蛋白表达水平显著下降；PGE₂的分泌水平显著提高，PGF_2α分泌水平显著下降。本研究明确了SRIF在绵羊子宫内膜组织和上皮细胞中广泛表达，在绵羊子宫内膜上皮细胞中加入SRIF-14能够抑制细胞增殖，诱导细胞发生凋亡。

关键词: 生长抑素, 绵羊子宫内膜上皮细胞, 增殖, 凋亡

Abstract: The purpose of this study was to explore the role of somatostatin release inhibitory factor (SRIF) in sheep endometrial epithelial cells. The uterine tissue samples of sheep were took from the designated slaughterhouse of cattle and sheep in Shihezi city, Xinjiang. The expression of SRIF in the endometrium of sheep was determined by immunohistochemical staining. Estradiol (E₂), progesterone (P₄) and interferon-tau (IFNT) were used to simulate the hormone changes during embryo implantation in sheep endometrium. RT-qPCR and ELISA were used to detect the expression of progestogen and prostaglandins (PGs), and Western blot was used to detect the expression level of SRIF. SRIF-14 was added to sheep endometrial epithelial cells, and cell proliferation was detected by CCK-8 kit and RT-qPCR. The protein expression levels of Caspase 9, Bax, Bcl-2 and Cyt-c were detected by Western blot. The levels of PGE₂ and PGF_2α secreted by cells were detected by ELISA kit. Immunohistochemical staining results showed that SRIF was widely expressed in sheep endometrial tissues, and cell staining results confirmed that SRIF was expressed in sheep endometrial epithelial cells. The results of hormone combined treatment of sheep endometrial epithelial cells showed that the mRNA expression levels of ISG15, CXCL10, HOXA10 and RSAD2 were significantly increased, the secretion level of PGE₂ was increased, the secretion level of PGF_2α was decreased, and the protein expression level of SRIF was significantly increased after hormone treatment. The results of exogenous addition of SRIF-14 in sheep endometrial epithelial cells showed that SRIF-14 inhibited the proliferation of sheep endometrial epithelial cells and the expression level of CCNA mRNA was significantly decreased. The expression level of Bax, Caspase 9 and Cyt-c proteins were significantly increased, and the expression level of Bcl-2 protein was significantly decreased. The secretion level of PGE₂ increased and the secretion level of PGF_2α decreased. This study confirmed that SRIF is widely expressed in sheep endometrial tissues and epithelial cells. The addition of SRIF-14 to sheep endometrial epithelial cells can inhibit cell proliferation and induce apoptosis.

Key words: SRIF, sheep endometrial epithelial cells, proliferation, apoptosis

中图分类号:

S826.3

宋美君, 郝科兴, 海思妤, 陈岩, 王静, 胡广东. SRIF-14对绵羊子宫内膜上皮细胞增殖和凋亡的影响[J]. 畜牧兽医学报, 2023, 54(8): 3325-3334.

SONG Meijun, HAO Kexing, HAI Siyu, CHEN Yan, WANG Jing, HU Guangdong. Effects of SRIF-14 on Proliferation and Apoptosis of Endometrial Epithelial Cells in Sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3325-3334.

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