与脐带间充质干细胞共培养对乳腺上皮细胞乳脂合成及关键基因表达的影响

doi:10.11843/j.issn.0366-6964.2017.05.008

摘要/Abstract

摘要：

旨在探究奶牛乳腺上皮细胞（BMECs）与脐带间充质干细胞（UC-MSCs）共培养对（BMECs）乳脂合成及关键基因表达的影响。试验共分为8组：共培养组为UC-MSCs和BMECs共培养条件下的不处理组、IGF-1 R抑制剂AG1024处理组、Janus激酶和转录活化因子（JAK/STAT）信号通路信号阻断剂AG490处理组及AG1024+AG490处理组，对照组为BMECs单培养条件下的不处理组、IGF-1 R抑制剂AG1024组、Janus激酶和转录活化因子（JAK/STAT）信号通路信号阻断剂AG490组及AG1024+AG490处理组。检测各组上清IGF-1、甘油三酯（TG）含量变化；RT-qPCR检测乙酰辅酶A羧化酶（ACACA），脂肪酸合成酶（FASN）和固醇调节元件结合蛋白（Sterol regulatory element binding proteins， SREBP 1）基因的相对表达丰度。结果表明，共培养组IGF-1、TG含量均显著高于对照组（P<0.05）；AG1024处理对IGF-1具有极显著抑制效果（P<0.01），显著降低TG含量及ACACA、FASN、SREBP1 mRNA相对表达丰度（P<0.05）；AG490处理对ACACA、FASN、SREBP 1 mRNA的表达无显著影响（P>0.05）；AG1024和AG490共同处理较AG1024单独处理各项指标表现差异不显著（P>0.05）。综上表明，脐带间充质干细胞能够通过IGF-1促进乳腺上皮细胞乳脂合成及关键基因的表达，JAK2/STAT5信号通路不参与脐带间充质干细胞对乳腺上皮细胞乳脂调控。

Abstract:

In order to explore the effect on milk fat synthesis and the expression of key genes of milk fat synthesis in bovine mammary gland epithelial cells co-cultured with umbilical cord mesenchymal stem cells, 8 groups were divided in the study: UC-MSCs and BMECs co-cultured under the condition of no treatment group, IGF-1 R inhibitor AG1024 treatment group, Janus kinase activators of transcription (JAK/STAT) pathway inhibitor AG490 treatment group and AG1024+AG490 treatment group, control group: BMECs alone cultured under the condition of no treatment group, IGF-1 R inhibitor AG1024 group, Janus kinase and transcription activating factor (JAK/STAT) signal pathway AG490 group and AG1024+AG490 treatment group. The IGF-1, Triglyceride (TG) content of each group supernatant were determined, the relative expression abundance of Acetyl-CoA carboxylase A (ACACA), fatty acid synthetase (FASN) and sterol regulatory element binding proteins (SREBP 1) mRNA were detected by RT-PCR. Results showed that the IGF-1 and TG content of co-culture group were significantly higher than control group (P<0.05); AG1024 treatment had a extremely significant effect on inhibiting IGF-1, and the TG content and the relative expression abundance of ACACA, FASN, SREBP 1 mRNA were significantly decreased(P<0.05); AG490 had no significant effect on the relative expression abundance of ACACA, FASN, SREBP 1 mRNA (P>0.05); AG1024 and AG490 co-treatment were not significant difference for each index than AG1024 treatment alone. (P>0.05). In conclusion, umbilical cord mesenchymal stem cells can promote milk fat synthesis and the expression of key gene in bovine mammary gland epithelial cells via IGF-1, but JAK2/STAT5 signaling pathway is not involving in the regulation of umbilical cord mesenchymal stem cells on milk fat of bovine mammary gland epithelial cells.

中图分类号:

S823.9⁺1.2

赵艳坤, 邵伟, 王立文, 余雄. 与脐带间充质干细胞共培养对乳腺上皮细胞乳脂合成及关键基因表达的影响[J]. 畜牧兽医学报, 2017, 48(5): 846-853.

ZHAO Yan-kun, SHAO Wei, WANG Li-wen, YU Xiong. Effect of Co-culturing with Umbilical Cord Mesenchymal Stem Cells on Milk Fat Synthesis and Expression of Key Genes in Bovine Mammary Gland Epithelial Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(5): 846-853.

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