脐带间充质干细胞通过类胰岛素样生长因子-Ⅰ介导的Janus激酶/信号转导及转录活化因子信号通路抑制奶牛乳腺上皮细胞凋亡的体外研究  

An in Vitro Study of Umbilical Cord Mesenchymal Stem Cells Inhibite Bovine Mammary Gland Epithelial Cells Apoptosis by Insulin Like Growth Factor-Ⅰ Mediated Janus Kinase/Signal Transducer and Activator of Transcription Signaling Pathway

作　　者：赵艳坤[1] 邵伟[1] 雒诚龙 武开乐 余雄[1] 

机构地区：[1]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,乌鲁木齐830052

出　　处：《动物营养学报》2017年第4期1335-1342,共8页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家奶产业体系(CARS-37);2015年国家自然科学基金项目(31560645);新疆农业大学畜牧学博士后工作站;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);新疆肉乳用草食动物营养实验室开放课题

摘　　要：本试验旨在探究Janus激酶/信号转导及转录活化因子(JAK/STAT)信号通路是否参与脐带间充质干细胞(UC-MSCs)通过类胰岛素样生长因子-Ⅰ(IGF-Ⅰ)抑制奶牛乳腺上皮细胞(BMECs)凋亡的调节。将UC-MSCs和BMECs利用TranswellTM小室双层共培养,以BMECs单纯培养为对照,给予类胰岛素样生长因子-Ⅰ受体(IGF-ⅠR)抑制剂AG1024进行干预,并用信号阻断剂AG490处理细胞,24 h后采用实时荧光定量PCR检测各组细胞B细胞淋巴瘤/白血病-2(Bcl-2)、B细胞淋巴瘤/白血病基因伴随蛋白x(Bax)、半胱氨酸蛋白酶3(Caspase-3)基因的相对表达丰度,流式细胞仪检测细胞凋亡情况。结果表明:UC-MSCs和BMECs共培养组BMECs的凋亡率极显著低于其他各组(P<0.01);UC-MSCs和BMECs共培养组Bcl-2基因的相对表达丰度较BMECs组极显著上调(P<0.01),Caspase-3、Bax基因的相对表达丰度则显著或极显著下调(P<0.05或P<0.01);AG1024和AG490单独处理或二者共同处理升高了单独培养的BMECs和与UC-MSCs共培养的BMECs的凋亡率,并上调了Bax、Caspase-3基因的相对表达丰度,下调了Bcl-2基因的相对表达丰度,均具有统计学意义(P<0.05或P<0.01)。由此得出,UC-MSCs能够通过IGF-Ⅰ介导JAK/STAT信号通路调节BMECs凋亡相关基因的表达,降低BMECs的凋亡率。This experiment was conducted to explore whether Janus kinase（ JAK）/signal transducer and activator of transcription（ STAT） signaling pathway was involved in regulation of umbilical cord mesenchymal stem cells（ UC-MSCs） by insulin like growth factor-Ⅰ（ IGF-Ⅰ） inhibited bovine mammary gland epithelial cells（ BMECs） apoptosis.UC-MSCs and BMECs were co-cultured by TranswellT Mdouble chamber,BMECs cultured alone as control,cells was treated with insulin like growth factor-Ⅰ receptor（ IGF-Ⅰ R） inhibitor AG1024 and signal blocking agent AG490.After 24 h,the relative expression abundances of B-cell lymphoma/leukemia-2（ Bcl-2）,Bcl-associated x protein（ Bax）,cysteine aspartic acid specific protease（ Caspase-3）were detected by real-time quantitative PCR（ RT-q PCR）,and flowcytometry was adopted to detect apoptosis changes of cells.The results showed as follows： the apoptosis rate of BMECs in UC-MSCs and BMECs co-culture group was significantly lower than that in other groups（ P〈0.01）; compared with BMECs group,the relative expression abundance of Bcl-2 gene in UC-MSCs and BMECs co-culture group was significantly increased（ P〈0.01）,while the relative expression abundances of Caspase-3 and Bax genes were significantly decreased（ P〈0.05 or P〈0.01）.Treated by AG1024 or/and AG490,the apoptosis rate of single cultured BMSCs and co-cultured BMSCs with UC-MSCs was raised,and the relative expression abundances of Bax and Caspase-3 genes were up-regulated,while the relative expression abundance of Bcl-2 gene was down-regulated,and they were statistically significant（ P〈0.05 or P〈0.01）.In conclusion,UC-MSCs can regulate the expression of apoptosis relation genes in BMECs by IGF-Ⅰ mediated JAK/STAT signaling pathway,and reduce the apoptosis rate of BMECs.

关 键 词：脐带间充质干细胞 乳腺上皮细胞 共培养 IGF-Ⅰ JAK/STAT 凋亡 

分 类 号：S852.2[农业科学—基础兽医学]