| 胰岛素样生长因子-1(IGF-1)对小鼠卵丘细胞自噬与胞吞相关因子表达的影响 |
| |
| 引用本文： | 马 睿,潘阳阳,王 萌,李 秦,何翃闳,胡学权,赵 凌,王靖雷,崔 燕,余四九.胰岛素样生长因子-1(IGF-1)对小鼠卵丘细胞自噬与胞吞相关因子表达的影响[J].西北农业学报,2019,28(9):1394-1402. |
| |
| 作者姓名： | 马 睿 潘阳阳 王 萌 李 秦 何翃闳 胡学权 赵 凌 王靖雷 崔 燕 余四九 |
| |
| 作者单位： | (甘肃农业大学 动物医学院/甘肃省牛羊胚胎工程技术研究中心，兰州 730070) |
| |
| 基金项目： | 国家自然科学基金青年基金（31702311）；甘肃省省青年科技基金计划（17JR5RA141）；甘肃省重点研发项目（17YF1NA058）；甘肃农业大学动物医学院创新基金(JYCX-KX016)；甘肃农业大学伏羲青年英才基金(Gaufx-02Y10)；甘肃农业大学博士科研启动基金(GSAU-RCZX201701)。 |
| |
| 摘 要： | 卵丘细胞的质量对卵母细胞的生长发育至关重要,其自噬和胞吞可调控细胞生长及营养运输,以应对恶劣环境对卵母细胞质量的威胁。从促排处理的小鼠输卵管膨大部收集卵丘—卵母细胞复合体(Cumulus oocyte complexes, COCs),分离卵丘细胞进行体外培养,并添加0、50、100、150和200 ng/mL IGF-1处理细胞,分别提取细胞总RNA和总蛋白,实时荧光定量PCR检测各处理组 Atg5、 Beclin1、 LC3、 Cav1和 Cav2 mRNA的相对表达水平。免疫荧光定位不同蛋白在细胞中的表达,并结合蛋白免疫印迹(Western-blot,WB)测定不同质量浓度IGF-1处理组Atg5、Beclin1、LC3、Cav1和Cav2的蛋白表达水平,分析IGF-1对卵丘细胞自噬和胞吞的影响。结果显示:正常培养的卵丘细胞均可表达 Atg5、 Beclin1、 LC3、 Cav1、 Cav2 mRNA和蛋白,蛋白主要定位在细胞质。IGF-1可调控小鼠卵丘细胞Atg5、Beclin1、LC3、Cav1和Cav2蛋白的表达,与对照组相比,各处理组的Atg5蛋白表达量均升高;Beclin1和LC3蛋白表达量显著降低,且在100 ng/mL IGF-1处理组中最低。Cav1和Cav2蛋白在50 ng/mL和200 ng/mL处理组的表达量均显著降低,但在该质量浓度变化范围内,随着IGF-1质量浓度增大,2种蛋白的表达量不是线性降低,而是在100 ng/mL和150 ng/mL处理组表达量有回升现象。表明IGF-1可调控小鼠卵丘细胞的自噬和胞吞相关因子的表达,调控效果与IGF-1的质量浓度有关。
|
| 关 键 词： | 胰岛素样生长因子-1 自噬 胞吞 卵丘细胞 |
Effects of Insulin-like Growth Factors IGF-1 on Factors Expression Related to Autophagy and Endocytosis in Mouse Cumulus Cells |
| |
| MA Rui,PAN Yangyang,WANG Meng,LI Qin,HE Honghong,HU Xuequan,ZHAO Ling,WANG Jinglei,CUI Yan and YU Sijiu.Effects of Insulin-like Growth Factors IGF-1 on Factors Expression Related to Autophagy and Endocytosis in Mouse Cumulus Cells[J].Acta Agriculturae Boreali-occidentalis Sinica,2019,28(9):1394-1402. |
| |
| Authors: | MA Rui PAN Yangyang WANG Meng LI Qin HE Honghong HU Xuequan ZHAO Ling WANG Jinglei CUI Yan and YU Sijiu |
| |
| Abstract: | The quality of cumulus cells is crucial for the growth and development of oocytes. Autophagy and endocytosis can regulate cell growth and nutrient transport to cope with the threat of oocyte quality in harsh environments. Cumulus oocyte complexes (COCs) were collected from the oviductal enlargement of the mouse, and the cumulus cells were isolated and cultured in vitro, and 0, 50 ng/mL, 100 ng/mL, 150 ng/mL and 200 ng/mL IGF-1were added to treat cells. Extracted total RNA and total protein, and detected the relative mRNA of Atg5, Beclin1, LC3, Cav1 and Cav2 in each treatment group by Quantitative Real-time PCR (qRT-PCR). Immunofluorescence technology localized the expression of different proteins in cells and the expression levels of Atg5,Beclin1, LC3,Cav1 and Cav2 in different concentrations of IGF-1 were detected by Western-blot (WB) technique. The effects of IGF-1 on autophagy and endocytosis of cumulus cells were analyzed. The results showed that normal cultured cumulus cells could express Atg5, Beclin1, LC3, Cav1 and Cav2 mRNA and protein, and their proteins were mainly located in the cytoplasm. IGF-1 regulated the expression of Atg5,Beclin1, LC3,Cav1 and Cav2 proteins in mouse cumulus cells. Compared with the control group, the expression of Atg5 protein was increased in each treatment group;and the expression levels of Beclin1 and LC3 proteins were significantly decreased and the expression levels were lowest in the 100 ng/mL IGF-1 treated group. The expression levels of Cav1 and Cav2 protein were significantly decreased in the 50 ng/mL and 200 ng/mL treatment groups.Within the range of mass concentration variation, the expression levels of the two proteins did not decrease linearly with the increase of IGF-1 concentration. Rather, the expression levels in the 100 ng/mL and 150 ng/mL treatment groups rebounded. The results indicated that IGF-1 can regulate the expression of autophagy and endocytosis-related factors in mouse cumulus cells, and its regulation effect is related to the mass concentration of IGF-1. |
| |
| Keywords: | Insulin-like growth factor-1 Autophagy Endocytosis Cumulus cells |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《西北农业学报》浏览原始摘要信息 |
| 点击此处可从《西北农业学报》下载免费的PDF全文 |
|
