与脐带间充质干细胞共培养促进奶牛乳腺上皮细胞乳蛋白合成及其机制

发布时间：2018-03-01 09:10

本文关键词： 脐带间充质干细胞(UCMSC) 牛乳腺上皮细胞(BMEC) 共培养乳蛋白 β酪蛋白(CSN) κ酪蛋白(CSN)　出处：《细胞与分子免疫学杂志》2017年02期 　论文类型：期刊论文

【摘要】：目的研究脐带间充质干细胞(UCMSC)调节奶牛乳腺上皮细胞(BMEC)乳蛋白合成的可能作用机制。方法利用TranswellTM小室将UCMSC和BMEC进行双层共培养,BMEC单纯培养为对照组,胰岛素样生长因子1受体(IGF-1R)抑制剂AG1024处理细胞,ELISA检测上清IGF-1和β酪蛋白(CSN2)、κ酪蛋白(CSN3)含量变化,实时定量PCR检测Janus激酶2/信号转导子与转录激活子5(JAK2/STAT5)信号通路相关基因的相对表达丰度;再用JAK2信号通路阻断剂AG490孵育细胞,实时定量PCR检测CSN2、CSN3 mRNA的相对表达丰度。结果与UCMSC共培养后,BMEC的CSN2、CSN3合成量和CSN2、CSN3、JAK2、STAT5、E74样ETS转录因子5(ELF5)mRNA相对表达丰度均显著高于单纯培养的BMEC;AG1024处理后,显著降低BMEC的CSN2、CSN3合成量,显著降低CSN2、CSN3、JAK2、STAT5、ELF5 mRNA的相对表达丰度;给予AG490阻断后,显著降低CSN2、CSN3 mRNA相对表达丰度;在AG1024基础上,加入AG490后显著降低CSN2、CSN3 mRNA相对表达丰度。结论UCMSC能够通过IGF-1介导JAK2/STAT5信号通路,上调BMEC乳蛋白合成关键基因mRNA的表达丰度,促进其乳蛋白的合成。
[Abstract]:Objective to study the possible mechanism of UCM SCC regulating milk protein synthesis of bovine mammary epithelial cells (BMECs). Methods UCMSC and BMEC were co-cultured in TranswellTM chamber as control group. Insulin like growth factor 1 receptor IGF-1R inhibitor (AG1024) was used to detect the changes of IGF-1 and 尾 casein (CSN2), 魏 casein (CSN3) levels in supernatants. Real-time quantitative PCR was used to detect the relative expression abundance of Janus kinase 2 / signal transducers associated with transcription activator 5 (JAK2 / STAT5) signaling pathway, and then incubated cells with AG490, a JAK2 signaling pathway blocker. Results after co-culture with UCMSC, the relative expression abundance of CSN2CN3 in BMEC and the relative expression of ETS transcription factor 5 ELF5N3 in CSN2CSN3- JAK2STAT5E74 were significantly higher than that in ECBMN1024 treated with UCMSC alone, and the synthesis of CSN2CSN3 in BMEC was significantly lower than that in ECBMN2A1024 treated with CSN2STAT5E74.Results the relative expression abundance of CSN2CSN3 in BMEC was significantly lower than that in cultured ECBMG1024, and the relative abundance of ETS transcription factor 5 ELF5N was significantly higher than that of ECBMG AG1024. After blocking AG490, the relative expression abundance of CSN2 mRNA was significantly decreased, and the relative abundance of mRNA expression was significantly decreased after the addition of AG4 90 on the basis of AG1024. Conclusion UCMSC can mediate the JAK2/STAT5 signaling pathway by IGF-1. Upregulate the expression abundance of mRNA, the key gene of BMEC milk protein synthesis, and promote its milk protein synthesis.
【作者单位】：新疆农业大学动物科学学院新疆肉乳用草食动物营养实验室;
【基金】：奶产业体系资助(CARS-37) 国家自然科学基金(31560645) 新疆农业大学畜牧学博士后工作站资助新疆维吾尔自治区高等学校科研计划项目(XJEDU2013S17) 2013年度新疆研究生科研创新项目(xjau-2013-yjsky-XJGRI2013113)
【分类号】：S823

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