IGF-Ⅰ对奶牛乳腺上皮细胞增殖和细胞周期的影响
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摘要
研究添加不同浓度的外源胰岛素样生长因子Ⅰ(IGF-Ⅰ)对奶牛乳腺上皮细胞(BMECs)增殖的影响。试验分为对照组和试验组,对照组仅添加与试验组等体积的完全培养基,试验组添加分别含25、50、75μg/mL IGF-Ⅰ的完全培养基,采用细胞计数试剂盒与流式细胞仪检测细胞在0、12、24、48、72 h时IGF-Ⅰ对细胞增殖和细胞周期的影响。结果表明,添加25μg/mL IGF-Ⅰ,培养12 h细胞增殖极显著高于对照组及其他试验组(P<0.01)。细胞培养12 h后,对照组细胞增殖显著高于试验组(P<0.05)。培养12 h添加IGF-Ⅰ缩短了细胞周期(P<0.01)。说明IGF-Ⅰ对细胞增殖具有一定的促进作用,可以缩短细胞周期。在培养基中添加25μg/mL IGF-Ⅰ,培养12 h,对奶牛乳腺上皮细胞的增殖作用最佳。
To study the effects of exogenous insulin-like growth factor-Ⅰ(IGF-Ⅰ) on the proliferation of bovine mammary gland epithelial cells(BMECs). The experiment was divided into the control group and the experimental group. The same volume of complete culture medium was added to the control group as that added to the experimental group. The experimental group was added the concentrations of 25,50,75 μg/mL of IGF-Ⅰ. The effects of cell proliferation and cell cycle at 0,12,24,48 and 72 h were detected by cell counting kit and flow cytometry. The results showed that the cell proliferation at 12 h was significantly higher when 25 μg/mL was added than that in the control group and other experimental groups(P<0.01). 12 hours later,the cell proliferation of the control group was significantly higher than that of the experimental group(P<0.05). The addition of IGF-Ⅰ at 12 h decreased the cell cycle(P<0.01). These results suggest that IGF-Ⅰ can promote cell proliferation and shorten cell cycle and the optimal concentration of IGF-Ⅰ was 25 μg/mL for 12 hours.
To study the effects of exogenous insulin-like growth factor-Ⅰ(IGF-Ⅰ) on the proliferation of bovine mammary gland epithelial cells(BMECs). The experiment was divided into the control group and the experimental group. The same volume of complete culture medium was added to the control group as that added to the experimental group. The experimental group was added the concentrations of 25,50,75 μg/mL of IGF-Ⅰ. The effects of cell proliferation and cell cycle at 0,12,24,48 and 72 h were detected by cell counting kit and flow cytometry. The results showed that the cell proliferation at 12 h was significantly higher when 25 μg/mL was added than that in the control group and other experimental groups(P<0.01). 12 hours later,the cell proliferation of the control group was significantly higher than that of the experimental group(P<0.05). The addition of IGF-Ⅰ at 12 h decreased the cell cycle(P<0.01). These results suggest that IGF-Ⅰ can promote cell proliferation and shorten cell cycle and the optimal concentration of IGF-Ⅰ was 25 μg/mL for 12 hours.
引文
[1] 邢艳苹.IGF-Ⅰ和HGF对奶牛乳腺上皮细胞合成乳脂乳蛋白的调节IGF-Ⅰ[D].长春:吉林大学,2012.
[2] Yan-Kun Z,Wei S,Cheng-Long L,et al.Proliferation-promoting effect of umbilical cord mesenchymal stem cells on co-cultured bovine mammary gland epithelial cells[J].Acta Laboratorium Animalis Scientia Sinica,2017,25(4):391-398.
[3] Rose M T,Weekes T E,Rowlinson P.Correlation of blood and milk components with the milk yield response to bovine somatotropin in dairy cows [J].Domestic Animal Endocrinology,2005,28(3):296-307.
[4] Burgos S A,Cant J P.IGF-1 stimulates protein synthesis by enhanced signaling through mTORC1 in bovine mammary epithelial cells[J].Domestic Animal Endocrinology,2010,38(4):211-221.
[5] 罗锐,张瑞智,龚正林,等.IGF-1对体外培养髁突软骨细胞增殖与凋亡的影响[J].实用口腔医学杂志,2016,32(6):810-814.
[6] 陈建晖,佟慧丽,李庆章,等.奶牛乳腺上皮细胞系的建立[J].畜牧兽医学报,2009,40(5):743-747.
[7] 薛亚梅,李坤,吕杰强.IGF-1对细胞凋亡的抑制调控[J].生命科学,2007,19(1):68-72.
[8] Curi R A,Oliveira H N,Silveira A C,et al.Effects of polymorphic microsatellites in the regulatory region of IGF1 and GHR on growth and carcass traits in beef cattle [J].Animal Genetics,2005,36(1):58-62.
[9] 何伯萍,邢艳苹,雷连成,等. IGF-Ⅰ对奶牛乳腺上皮细胞合成乳脂、乳蛋白的影响[J].中国兽医学报,2012,32(8):1231-1234.
[10] 吴云娣.牛奶中IGF-Ⅰ含量调查及其原核表达与活性分析[D].长春:吉林大学,2013.
[11] Manjarín R,Steibel J P,Kirkwood R N,et al.Transcript abundance of hormone receptors,mammalian target of rapamycin pathway-related kinases,insulin-like growth factor I,and milk proteins in porcine mammary tissue[J].Journal of Animal Science,2012,90(1):221-230.
[12] 高玉红,李庆章.IGFs对小鼠乳腺发育及细胞凋亡的影响[J].中国乳品工业,2007,35(12):26-28.
[13] 莫继先,佟慧丽,张莉,等.牛初乳IGF-I对体外培养人乳腺上皮细胞系HBL-100影响的研究[J].药物生物技术,2008,15(6):431-434.
[14] 张童非.奶牛乳腺上皮细胞胰岛素样生长因子及受体IGF-IR的表达及作用[D].哈尔滨:东北农业大学,2012.
[15] 李妍妍,张辉,秦晓毅,等.PI3K/Akt信号通路激活抑制细胞凋亡的研究进展[J].浙江临床医学,2013(4):574-577.
[16] 雒诚龙,赵艳坤,武开乐,等.添加通路阻断剂对IGF-1影响奶牛乳腺上皮细胞增殖与凋亡的研究[J].中国畜牧兽医,2017,44(8):2277-2282.
[17] Lochrie J D,Phillips K,Tonner E,et al.Insulin-like growth factor binding protein (IGFBP)-5 is upregulated during both differentiation and apoptosis in primary cultures of mouse mammary epithelial cells[J].Journal of Cellular Physiology,2006,207(2):471–479.
[18] 高玉红,李庆章.IGFs对小鼠乳腺IGFBP-5和cyclin D1表达的影响[J].中国乳品工业,2007,35(11):41-43.
[19] 季昀.GH和IGF-I对奶牛乳腺上皮细胞中酪蛋白合成的调节作用机制[D].扬州大学,2013.
[20] 李莹莹,夏小静,吴云娣,等.胰岛素样生长因子-Ⅰ、肝细胞生长因子、转化生长因子-β1、干扰素-γ对奶牛乳腺上皮细胞增殖活性的影响[J].中国畜牧兽医,2014,41(4):37-42.
[21] 岑石强,张峻梅,黄富国,等.IGF-1促原代人胚骨骼肌成肌细胞体外增殖与分化的研究[J].中国修复重建外科杂志,2008,22(1):84-87.
[22] 秦君,李庆章,高学军.王不留行主要成分对小鼠乳腺上皮细胞增殖及β-酪蛋白表达的影响[J].中国农业科学,2008,41(8):2442-2447.
[23] 金贞爱,公明花,赵一坤,等.胰岛素样生长因子-1对高氧暴露的A549细胞周期的影响[J].中国妇幼保健,2014,29(12):1943-1946.
[24] 邹向阳,李连宏.细胞周期调控与肿瘤[J].国际遗传学杂志,2006,29(1):70-73.
[25] 张艳平,丁矢,李雪兰,等.shRNA表达载体沉默BHRF1对鼻咽癌CNE2细胞生长、凋亡及细胞周期的影响[J].中国医药指南,2015(5):55-55.
[2] Yan-Kun Z,Wei S,Cheng-Long L,et al.Proliferation-promoting effect of umbilical cord mesenchymal stem cells on co-cultured bovine mammary gland epithelial cells[J].Acta Laboratorium Animalis Scientia Sinica,2017,25(4):391-398.
[3] Rose M T,Weekes T E,Rowlinson P.Correlation of blood and milk components with the milk yield response to bovine somatotropin in dairy cows [J].Domestic Animal Endocrinology,2005,28(3):296-307.
[4] Burgos S A,Cant J P.IGF-1 stimulates protein synthesis by enhanced signaling through mTORC1 in bovine mammary epithelial cells[J].Domestic Animal Endocrinology,2010,38(4):211-221.
[5] 罗锐,张瑞智,龚正林,等.IGF-1对体外培养髁突软骨细胞增殖与凋亡的影响[J].实用口腔医学杂志,2016,32(6):810-814.
[6] 陈建晖,佟慧丽,李庆章,等.奶牛乳腺上皮细胞系的建立[J].畜牧兽医学报,2009,40(5):743-747.
[7] 薛亚梅,李坤,吕杰强.IGF-1对细胞凋亡的抑制调控[J].生命科学,2007,19(1):68-72.
[8] Curi R A,Oliveira H N,Silveira A C,et al.Effects of polymorphic microsatellites in the regulatory region of IGF1 and GHR on growth and carcass traits in beef cattle [J].Animal Genetics,2005,36(1):58-62.
[9] 何伯萍,邢艳苹,雷连成,等. IGF-Ⅰ对奶牛乳腺上皮细胞合成乳脂、乳蛋白的影响[J].中国兽医学报,2012,32(8):1231-1234.
[10] 吴云娣.牛奶中IGF-Ⅰ含量调查及其原核表达与活性分析[D].长春:吉林大学,2013.
[11] Manjarín R,Steibel J P,Kirkwood R N,et al.Transcript abundance of hormone receptors,mammalian target of rapamycin pathway-related kinases,insulin-like growth factor I,and milk proteins in porcine mammary tissue[J].Journal of Animal Science,2012,90(1):221-230.
[12] 高玉红,李庆章.IGFs对小鼠乳腺发育及细胞凋亡的影响[J].中国乳品工业,2007,35(12):26-28.
[13] 莫继先,佟慧丽,张莉,等.牛初乳IGF-I对体外培养人乳腺上皮细胞系HBL-100影响的研究[J].药物生物技术,2008,15(6):431-434.
[14] 张童非.奶牛乳腺上皮细胞胰岛素样生长因子及受体IGF-IR的表达及作用[D].哈尔滨:东北农业大学,2012.
[15] 李妍妍,张辉,秦晓毅,等.PI3K/Akt信号通路激活抑制细胞凋亡的研究进展[J].浙江临床医学,2013(4):574-577.
[16] 雒诚龙,赵艳坤,武开乐,等.添加通路阻断剂对IGF-1影响奶牛乳腺上皮细胞增殖与凋亡的研究[J].中国畜牧兽医,2017,44(8):2277-2282.
[17] Lochrie J D,Phillips K,Tonner E,et al.Insulin-like growth factor binding protein (IGFBP)-5 is upregulated during both differentiation and apoptosis in primary cultures of mouse mammary epithelial cells[J].Journal of Cellular Physiology,2006,207(2):471–479.
[18] 高玉红,李庆章.IGFs对小鼠乳腺IGFBP-5和cyclin D1表达的影响[J].中国乳品工业,2007,35(11):41-43.
[19] 季昀.GH和IGF-I对奶牛乳腺上皮细胞中酪蛋白合成的调节作用机制[D].扬州大学,2013.
[20] 李莹莹,夏小静,吴云娣,等.胰岛素样生长因子-Ⅰ、肝细胞生长因子、转化生长因子-β1、干扰素-γ对奶牛乳腺上皮细胞增殖活性的影响[J].中国畜牧兽医,2014,41(4):37-42.
[21] 岑石强,张峻梅,黄富国,等.IGF-1促原代人胚骨骼肌成肌细胞体外增殖与分化的研究[J].中国修复重建外科杂志,2008,22(1):84-87.
[22] 秦君,李庆章,高学军.王不留行主要成分对小鼠乳腺上皮细胞增殖及β-酪蛋白表达的影响[J].中国农业科学,2008,41(8):2442-2447.
[23] 金贞爱,公明花,赵一坤,等.胰岛素样生长因子-1对高氧暴露的A549细胞周期的影响[J].中国妇幼保健,2014,29(12):1943-1946.
[24] 邹向阳,李连宏.细胞周期调控与肿瘤[J].国际遗传学杂志,2006,29(1):70-73.
[25] 张艳平,丁矢,李雪兰,等.shRNA表达载体沉默BHRF1对鼻咽癌CNE2细胞生长、凋亡及细胞周期的影响[J].中国医药指南,2015(5):55-55.