苜蓿黄酮对热应激下体外培养奶牛乳腺上皮细胞凋亡的影响

doi:10.11686/cyxb2015317

草业学报 ›› 2016, Vol. 25 ›› Issue (4): 159-165.DOI: 10.11686/cyxb2015317

苜蓿黄酮对热应激下体外培养奶牛乳腺上皮细胞凋亡的影响

占今舜¹, 魏明吉^{1, 2}, 苏效双¹, 詹康¹, 刘明美^{1, 3}, 张春刚^{1, 4}, 赵国琦^{1*, *}

1.扬州大学动物科学与技术学院,江苏扬州 225009;
2.临沂大学生命科学学院,山东临沂 276000;
3.江苏联合职业技术学院淮安生物工程分院,江苏淮安 223200;
4.上海光明荷斯坦牧业有限公司,上海 200443

收稿日期:2015-06-25 出版日期:2016-04-20 发布日期:2016-04-20
作者简介:占今舜(1985-),男,江西玉山人,在读博士。E-mail:zhanjinshun1985@163.com
基金资助:
江苏省高校优势学科建设工程项目(PAPD),长三角地区生鲜乳质量安全追溯体系关键技术研究应用项目(14395810100)和江苏省高校研究生科研创新计划项目(KYZZ_0367)资助

Effect of alfalfa flavonoids on bovine mammary epithelial cell cultures in vitro under heat stress

ZHAN Jin-Shun¹, WEI Ming-Ji^{1, 2}, SU Xiao-Shuang¹, ZHAN Kang¹, LIU Ming-Mei^{1, 3}, ZHANG Chun-Gang^{1, 4}, ZHAO Guo-Qi^{1, *}

1.College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;
2.College of Animal Science, Linyi University, Linyi 276000, China;
3.Jiangsu Joint Institute of Technology of Profession of Huai’an Bio-engineering Branch, Huai’an 223200, China;
4.Shanghai Bright Holstan Co., Ltd, Shanghai 200443, China

Received:2015-06-25 Online:2016-04-20 Published:2016-04-20

摘要/Abstract

摘要： 本实验旨在研究苜蓿黄酮对热应激下体外培养奶牛乳腺上皮细胞的影响。将乳腺上皮细胞分成5组,每组培养基中分别含有0,25,50,75和100 μg/mL苜蓿黄酮,同时置于细胞培养箱37℃,5%CO₂培养72 h,再在 42℃恒温水浴锅中热应激1 h后返回细胞培养箱培养12 h,检测细胞活性、抗氧化指标和相关基因的表达。结果显示:1)添加25 μg/mL组的细胞活性显著高于0和50 μg/mL组(P<0.05),其他各组之间差异不显著。2)相对于0 μg/mL组,50~100 μg/mL组细胞的GSH-Px活性升高(P<0.01),LDH和MDA含量降低(P<0.01或P<0.05),而CAT活性无显著性差异。3)相对于0 μg/mL组,50和75 μg/mL组Caspase3和Socs3基因表达降低(P<0.01),25 μg/mL组P53、Stat1和Socs1基因表达升高(P<0.01或P<0.05),而Bcl-2和Fas基因表达无显著差异。综上所述,在热应激下,苜蓿黄酮能够提高体外培养奶牛乳腺上皮细胞的活性,改善抗氧化能力和抑制细胞凋亡,其中添加75 μg/mL效果较好。

Abstract: The aim of this study was to evaluate the effect of alfalfa flavonoids on in vitro bovine mammary epithelial cells under heat stress. Bovine mammary epithelial cells were cultured on media with different concentrations of alfalfa flavonoids (0, 25, 50, 75, and 100 μg/mL) at 37℃ with 5% CO₂ for 72 h. Then, the cultured cells were subjected to a heat-shock treatment (42℃ for 1 h) before being returned to 37℃ with 5% CO₂ for 12 h. The cell activity, antioxidant index, and gene transcript levels were evaluated. The results showed that the cell activity was significantly higher in the 25 μg/mL treatment group than in the 0 and 50 μg/mL treatment groups (P<0.05), but cell activity did not differ significantly among the other groups. Compared with the control group (0 μg/mL) the 50, 75, and 100 μg/mL treatment groups showed significantly higher glutathione peroxidase activity (P<0.01) and significantly lower lactate dehydrogenase activity and malondialdehyde contents (P<0.01 and P<0.05, respectively). The catalase activity did not differ significantly among the five groups. Compared with the control group (0 μg/mL), the 50 and 75 μg/mL treatment groups showed reduced transcript levels of Caspase3 and Socs3, and the 25 μg/mL group showed significantly increased transcript levels of P53, Stat1, and Socs1 (P<0.01 or P<0.05). The transcript levels of Bcl-2 and Fas did not differ significantly among the groups. Together, these results show that alfalfa flavonoids can increase cell activity and antioxidant capacity and inhibit cell apoptosis, and that the optimal concentration of alfalfa flavonoids is 75 μg/mL.

占今舜, 魏明吉, 苏效双, 詹康, 刘明美, 张春刚, 赵国琦. 苜蓿黄酮对热应激下体外培养奶牛乳腺上皮细胞凋亡的影响[J]. 草业学报, 2016, 25(4): 159-165.

ZHAN Jin-Shun, WEI Ming-Ji, SU Xiao-Shuang, ZHAN Kang, LIU Ming-Mei, ZHANG Chun-Gang, ZHAO Guo-Qi. Effect of alfalfa flavonoids on bovine mammary epithelial cell cultures in vitro under heat stress[J]. Acta Prataculturae Sinica, 2016, 25(4): 159-165.

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苜蓿黄酮对热应激下体外培养奶牛乳腺上皮细胞凋亡的影响

Effect of alfalfa flavonoids on bovine mammary epithelial cell cultures in vitro under heat stress

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