基于2D-DIGE/MALDI-TOF-MS技术筛选奶牛低血钙症生物标志物

doi:10.11843/j.issn.0366-6964.2015.07.022

畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (7): 1238-1245.doi: 10.11843/j.issn.0366-6964.2015.07.022

基于2D-DIGE/MALDI-TOF-MS技术筛选奶牛低血钙症生物标志物

舒适，夏成^*，张洪友，许楚楚，李昌盛，肖鑫焕，王刚，白云龙

(黑龙江八一农垦大学动物科技学院，大庆 163319)

收稿日期:2014-11-02 出版日期:2015-07-23 发布日期:2015-07-23
通讯作者: 夏成，E-mail：xcwlxyf2014@163.com
作者简介:舒适(1986-)，女，浙江宁波人，博士生，主要从事临床兽医学的研究，E-mail：519296311@qq.com
基金资助:
国家自然科学基金项目主任基金（30840060）；国家自然科学基金面上项目（30972235）；黑龙江省自然科学基金面上项目（C200916）

Biomarker Discovery for Cows with Hypocalcemia Using 2D-DIGE

SHU Shi，XIA Cheng^*，ZHANG Hong-you，XU Chu-chu，LI Chang-sheng，XIAO Xin-huan，WANG Gang，BAI Yun-long

(College of Animal Science and Veterinary Medicine，Heilongjiang Bayi Agricultural University，Daqing 163319，China)

Received:2014-11-02 Online:2015-07-23 Published:2015-07-23

摘要/Abstract

摘要：

奶牛乳热（MF）是一种严重的营养代谢性疾病，通常被分为临床型及亚临床型低血钙症（SH）。奶牛乳热的诊断通常是应用奶牛血浆中的钙浓度进行判定的，但是在奶牛乳热整个发生发展的过程中，发病的具体机制仍是研究的盲点。在本试验中通过筛选奶牛乳热、亚临床低血钙症及健康奶牛间生物标志物进而解释可能的发生机制。选取27头奶牛作为实验动物，并根据其血浆钙浓度及有无临床症状分为乳热组（MF组）、亚临床低血钙组（SH组）和正常组（C组）。在组内每3个样品混合成为1个混合样本应用于2D-DIGE试验。结果得到110个差异蛋白质点，选取其中80个点进行MALDI-TOF-MS质谱分析，得到66个阳性结果并整合成为16种蛋白质。根据试验结果，选取A2M、HP和PON1进行Western blotting验证试验。本试验首次证实A2M、HP和PON1是奶牛乳热症3种新的生物标志物，并分析了其在乳热症发生过程中的可能作用机制。

Abstract:

Milk fever (MF) is a seriously nutrition metabolism disease which is divide into clinical and subclinical hypocalcemia (SH).The level of Ca in blood combining the clinical signal were always used to diagnose MF.But in the total development progress of this disease the pathogenesis was not clearly.In this study we screened the biomarkers of MF and SH，and want to reveal the possibly pathogenesis.Twenty-seven dairy cows were selected as the experiment animal，and there were 3 groups MF，SH and C according to the blood calcium concentration and the clinical signal of these dairy cows.Every 3 samples were mixed into a hybrid sample in each group，and 9 hybrid samples were crossed into the 2D-DIGE.Eighty of 110 spots，which were found via 2D-DIGE，were selected into mass spectrum.Sixty-six spots were found，and were identified into 16 proteins.A2M，HP，PON1 were selected in verification test by using Western blotting，and the results support the results of DIGE experiment.In this study 16 proteins were found among three groups，and 3 proteins of them were verified first time.Collectively，the results indicate that A2M，HP，PON1 are 3 novel biomarkers for MF，and they may contribute to explore the possible mechanism of MF.

中图分类号:

S857.2

舒适，夏成，张洪友，许楚楚，李昌盛，肖鑫焕，王刚，白云龙. 基于2D-DIGE/MALDI-TOF-MS技术筛选奶牛低血钙症生物标志物[J]. 畜牧兽医学报, 2015, 46(7): 1238-1245.

SHU Shi，XIA Cheng，ZHANG Hong-you，XU Chu-chu，LI Chang-sheng，XIAO Xin-huan，WANG Gang，BAI Yun-long. Biomarker Discovery for Cows with Hypocalcemia Using 2D-DIGE[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(7): 1238-1245.

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基于2D-DIGE/MALDI-TOF-MS技术筛选奶牛低血钙症生物标志物

Biomarker Discovery for Cows with Hypocalcemia Using 2D-DIGE

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