miR-221对奶牛乳腺上皮细胞乳脂、乳蛋白合成的影响及作用机制  

Effect of miR-221 on the synthesis of milk fat and milk protein in bovine mammary epithelial cells and its mechanism

作　　者：冉耀祥 魏祥飞 曲波[1] 王春梅[1] 姜毓君[1] 张莉[1] RAN Yaoxiang;WEI Xiangfei;QU Bo;WANG Chunmei;JIANG Yujun;ZHANG Li(Key Laboratory of Dairy Science,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]东北农业大学乳品科学教育部重点实验室,哈尔滨150030

出　　处：《中国乳品工业》2025年第1期26-31,共6页China Dairy Industry

基　　金：国家自然科学基金项目(31501033)。

摘　　要：文章利用体外分离培养奶牛乳腺上皮细胞研究miR-221对乳脂和乳蛋白合成的影响。将miR-221转染至奶牛乳腺上皮细胞中,应用QRT-PCR检测miR-221表达量,应用Western blot技术和组织细胞甘油三酯(TG)含量酶法分别检测β-酪蛋白和甘油三酯含量。利用双荧光素酶试验分析miR-221和IGF-1的靶向关系,检测转染miR-221后IGF-1和相关泌乳信号通路蛋白表达的变化。结果表明,在奶牛乳腺上皮细胞中转染miR-221后,miR-221表达显著上升,β-酪蛋白和甘油三酯含量显著降低;泌乳调节相关基因IGF-1与miR-221具有靶向关系,过表达miR-221后,IGF-1、AKT、p-AKT、mTOR、p-mTOR的表达也显著降低,表明miR-221可能通过靶向抑制IGF-1进而影响AKT/mTOR信号通路来影响乳脂和乳蛋白合成。In this study,the effects of miR-221 on milk fat and milk protein synthesis were investigated by using bovine mammary epithelial cells isolated and cultured in vitro.Firstly,miR-221 was transfected into bovine mammary epithelial cells,and the expression of miR-221 was detected by QRT-PCR.Western blotting and triglyceride(TG)content assay were used to detectβ-casein and TG content.Secondly,the targeting relationship between miR-221 and IGF-1 was analyzed by double luciferase assay,and the changes of IGF-1 and related lactation signaling pathway protein expression after transfection of miR-221 were detected.The results showed that the expression of miR-221 was significantly increased in bovine mammary epithelial cells transfected with miR-221,and the contents ofβ-casein and TG were significantly decreased.The IGF-1 gene related to lactation regulation had a targeting relationship with miR-221,and the expression of IGF-1,AKT,p-AKT,mTOR,and p-mTOR were significantly decreased after overexpression of miR-221,suggesting that miR-221 may affect milk fat and milk protein synthesis by targeting IGF-1 and then affecting the AKT/mTOR signaling pathway.

关 键 词：奶牛乳腺上皮细胞 MIR-221 IGF-1 乳脂 乳蛋白 

分 类 号：S823[农业科学—畜牧学]