金属硫蛋白对热应激下体外培养奶牛淋巴细胞的影响

doi:10.11686/cyxb20140325

草业学报 ›› 2014, Vol. 23 ›› Issue (3): 215-223.DOI: 10.11686/cyxb20140325

金属硫蛋白对热应激下体外培养奶牛淋巴细胞的影响

占今舜^1,2,吴力专¹,李丽立³,张彬^1,*

1.湖南农业大学动物科学技术学院,湖南长沙 410128;
2.扬州大学动物科学技术学院,江苏扬州 225009;
3.中国科学院亚热带农业生态研究所动物生态营养与健康养殖联合实验室农业生态工程重点实验室,湖南长沙 410125

收稿日期:2013-06-21 出版日期:2014-06-20 发布日期:2014-06-20
通讯作者: E-mail:zhb8236@126.com
作者简介:占今舜(1985-),男,江西玉山人,硕士。E-mail:zhanjs1023@sina.com
基金资助:
国家自然科学基金项目(31072053,30671516),湖南省自然科学基金重点项目(11JJ2014)和教育部高等学校博士学科点专项科研基金(博导类)项目(20104320110001)资助

Effect of metallothionein on lymphocyte apoptosis of the dairy cow cultured in vitro under heat stress

ZHAN Jin-shun^1,2,WU Li-zhuan¹,LI li-li³,ZHANG Bin¹

1.College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China;
2.College of Animal Science and Technology, Yangzhou University, yangzhou 225009, China;
3.Key Laboratory of Agro-ecology, Laboratory of Animal Nutrition and Human Health, Institute of Subtropical Agriculture, the Chinese Academy of Science, Changsha 410125, China

Received:2013-06-21 Online:2014-06-20 Published:2014-06-20

摘要/Abstract

摘要： 本研究的目的在于探讨外源性金属硫蛋白(MT)对热应激下体外培养奶牛淋巴细胞凋亡的影响。将屠宰获取的奶牛脾脏淋巴细胞随机分为4组,每组细胞培养液中MT的终浓度分别为0 μg/mL(对照组)、70 μg/mL(第Ⅰ组)、140 μg/mL(第Ⅱ组)、210 μg/mL(第Ⅲ组),同时置于细胞培养箱(37℃、5%CO₂)中培养20 h,之后同时置于43℃水浴锅中进行热应激处理1 h,再回细胞培养箱中培养3 h,然后利用形态学观察、RT-PCR等方法进行各项指标的检测。结果如下,1)通过形态学观察和AO/EB染色观察,表明添加MT可以有效缓解热应激对奶牛淋巴细胞的损伤,但缓解效果随MT浓度的增加而下降; 2)第Ⅰ~Ⅲ组的CAT活性、SOD活性和GSH-Px活性均高于对照组(P<0.01),MDA含量均低于对照组(P<0.01),表明MT能够有效改善淋巴细胞的抗氧化能力;3)试验Ⅰ~Ⅲ组的Bax基因相对表达量均显著低于对照组(P<0.01),但试验Ⅰ~Ⅲ组Bcl-2基因的表达量和p50基因的表达量均显著低于对照组(P<0.01),表明MT可以调节与细胞凋亡相关基因的表达。综上所述,添加MT可以降低奶牛淋巴细胞死亡率,改善奶牛淋巴细胞抗氧化性能,抑制奶牛淋巴细胞凋亡,以添加终浓度为70 μg/mL的MT效果最好。

Abstract: The effect of exogenous MT on lymphocyte apoptosis of the dairy cow was studied in vitro under heat stress. The splenic dairy cow lymphocytes were divided into 4 groups which had final concentrations of MT 0 (control group), 70 (group Ⅰ), 140 (group Ⅱ) and 210 μg/mL (group Ⅲ). The cells of each group were cultured in a cell incubator (37℃, 5%CO₂). After 20 h, the cells of each group were put into water bath pot (43℃) for 1 h, and then recultured in a cell incubator (37℃, 5%CO₂) for 3 h. Finally, various indexes were tested by observation of morphology and RT-PCR. 1) MT added in cell culture medium reduced the damage of dairy cow lymphocytes under heat stress, but the effect decreased with an increase in MT concentration. 2) The activities of CAT, SOD, GSH-Px in groups Ⅰ to Ⅲ were higher than those in the control group and the content of MDA were lower. It is suggested that MT could improve antioxidation of lymphocytes. 3) The relative transcript levels of Bax and p50 in group Ⅰ to Ⅲ were significantly (P<0.01) lower than those in the control group. The relative transcript level of Bcl-2 in groups Ⅰ to Ⅲ were significantly (P<0.01) higher than those in the control group. This indicated that MT could regulate the expression of genes related to lymphocyte apoptosis. In conclusion, exogenous MT decreased lymphocyte mortality rate, improved lymphocyte inoxidizability and restrained lymphocyte apoptosis . The best final concentration of MT was 70 μg/mL.

中图分类号:

S823.9⁺1

占今舜,吴力专,李丽立,张彬. 金属硫蛋白对热应激下体外培养奶牛淋巴细胞的影响[J]. 草业学报, 2014, 23(3): 215-223.

ZHAN Jin-shun,WU Li-zhuan,LI li-li,ZHANG Bin. Effect of metallothionein on lymphocyte apoptosis of the dairy cow cultured in vitro under heat stress[J]. Acta Prataculturae Sinica, 2014, 23(3): 215-223.

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金属硫蛋白对热应激下体外培养奶牛淋巴细胞的影响

Effect of metallothionein on lymphocyte apoptosis of the dairy cow cultured in vitro under heat stress

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