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 acti-vator 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 TranswellTM double chamber, BMECs cultured alone as control, cells was treated with insulin like growth factor-Ⅰ receptor ( IGF-ⅠR ) inhibitor AG1024 and signal blocking agent AG490. After 24 h, the relative expression abundances of B-cell lympho-ma/leukemia-2 ( Bcl-2) , Bcl-associated x protein ( Bax) , cysteine aspartic acid specific protease ( Caspase-3) were detected by real-time quantitative PCR ( RT-qPCR) , and flow cytometry was adopted to detect apoptosis changes of cells. The results showed as follows:the apoptosis rate of BMECs in UC-MSCs and BMECs co-cul-ture group was significantly lower than that in other groups ( P<0.01);compared with BMECs group, the rel-ative 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 ap-optosis relation genes in BMECs by IGF-Ⅰ mediated JAK/STAT signaling pathway, and reduce the apoptosis rate of BMECs.
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