miR-let-7a对脂磷壁酸诱导炎性牛乳腺上皮细胞增殖和凋亡的作用  

Effect of miR-let-7a on proliferation and apoptosis of inflammatory bovine mammary epithelial cells induced by lipoteichoic acid

作　　者：徐萍 炊寒冰 王彤 申祥 张晓建[2] XU Ping;CHUI Han-bing;WANG Tong;SHEN Xiang;ZHANG Xiao-jian(School of Biological Engineering,Xinxiang University/Animal Disease Molecular Diagnostic of Henan Engineering Laboratory,Xinxiang,Henan 453003,China;College of Animal Science and Technology,Henan Institute of Science and Technology,Xinxiang,Henan 453003,China)

机构地区：[1]新乡学院生物工程学院/动物疫病分子诊断河南省工程实验室,河南新乡453003 [2]河南科技学院动物科技学院,河南新乡453003

出　　处：《西南农业学报》2024年第7期1622-1627,共6页Southwest China Journal of Agricultural Sciences

基　　金：河南省国际科技合作项目(232102520014);河南省高等学校重点科研项目(23A180025、21A230002);河南省科技攻关项目(222102110046)。

摘　　要：[目的]研究微小RNA let-7a(microRNA let-7a,miR-let-7a)对脂磷壁酸(Lipoteichoic acid,LTA)诱导的炎性奶牛乳腺上皮细胞(MAC-T)的增殖和凋亡作用。[方法]通过实时荧光定量PCR(Quantitative real time polymerase chain reaction,qRT-PCR)、实时细胞分析仪和流式细胞术检测过表达和抑制表达miR-let-7a在炎性MAC-T中的影响。[结果]qRT-PCR检测结果显示,转染miR-let-7a模拟物会使炎性MAC-T细胞中miR-let-7a的表达量极显著上调(P<0.001),转染miR-let-7a抑制剂会使细胞中miR-let-7a的表达量极显著下降(P<0.001);RTCA结果显示,转染miR-let-7a模拟物能明显促进MAC-T细胞增殖,转染miR-let-7a抑制物会使MAC-T细胞的增殖受到抑制;流式细胞术结果显示,转染miR-let-7a模拟物的MAC-T细胞的凋亡率显著降低(P<0.01);而转染miR-let-7a抑制剂则与之相反,MAC-T细胞的凋亡率显著升高(P<0.01)。即过表达miR-let-7a可以促进MAC-T细胞增殖,并降低细胞的凋亡率。而抑制表达miR-let-7a的作用与之相反,抑制表达miR-let-7a使MAC-T细胞的增殖下降,细胞的凋亡率增大。[结论]miR-let-7a是一种抑制炎症调节因子,其过表达可以缓解奶牛乳腺炎,为阐明奶牛乳腺炎的分子调控机制奠定基础。[Objective]The study aimed to investigate the effect of microRNA let-7a(miR-let-7a)on the proliferation and apoptosis of inflam-matory bovine mammary epithelial cells(MAC-T)induced by lipoteichoic acid(LTA).[Method]miR-let-7a was over-expressed and inhibi-ted on inflammatory MAC-T,and its function was detected by Quantitative real time polymerase chain reaction(qRT-PCR),ANNEXIN V-FITC/PI,and real time cell analysis.[Result]The qRT-PCR results showed that transfection of miR-let-7a mimetic significantly upregulated the expression of miR-let-7a in inflammatory MAC-T cells(P<0.001),while transfection of miR-let-7a inhibitor significantly decreased the expression of miR-let-7a in MAC-T cells(P<0.001).The RTCA results showed that transfection of miR-let-7a mimetic significantly pro-moted the proliferation of MAC-T cells,while transfection of miR-let-7a inhibitors inhibited the proliferation of MAC-T cells.And the results of flow cytometry showed that the apoptosis rate of MAC-T cells transfected with miR-let-7a mimetic was significantly reduced(P<0.01);On the contrary,transfection with miR-let-7a inhibitor significantly increased the apoptosis rate of MAC-T cells(P<0.01).In summary,the results showed that over-expressed of miR-let-7a could promote the proliferation of MAC-T,and reduce the apoptosis rate of MAC-T cells,The effect of inhibiting the expression of miR-let-7a was opposite.Inhibiting the expression of miR-let-7a leaded to a decrease in prolifer-ation of MAC-T cells and an increase in cell apoptosis rate.[Conclusion]miR-let-7a is an anti-inflammatory regulator factor,and its over-ex-pression can alleviate bovine mastitis.These findings lay a foundation for elucidating the molecular regulation mechanism of bovine mastitis.

关 键 词：miR-let-7a 奶牛乳腺上皮细胞 炎症 脂磷壁酸 

分 类 号：S823[农业科学—畜牧学]