Lai Yu-Chang, Fujikawa Takuro, Maemura Tadashi, Ando Takaaki, Kitahara Go, Endo Yasuyuki, Yamato Osamu, Koiwa Masateru, Kubota Chikara, Miura Naoki
Veterinary Teaching Hospital, Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima, Japan.
The United Graduate School of Veterinary Science, Yamaguchi University, Yamaguchi, Japan.
PLoS One. 2017 May 17;12(5):e0177182. doi: 10.1371/journal.pone.0177182. eCollection 2017.
MicroRNA (miRNA) in tissue and liquid samples have been shown to be associated with many diseases including inflammation. We aimed to identify inflammation-related miRNA expression level in the bovine mastitis milk. Expression level of inflammation-related miRNA in milk from mastitis-affected and normal cows was analyzed using qPCR. We found that expression level of miR-21, miR-146a, miR-155, miR-222, and miR-383 was significantly upregulated in California mastitis test positive (CMT+) milk. We further analyzed these miRNA using a chip-based QuantStudio Digital PCR System. The digital PCR results correlated with those of qPCR, demonstrating upregulation of miR-21, miR-146a, miR-155, miR-222, and miR-383 in CMT+ milk. In conclusion, we identified miRNA that are upregulated in CMT+ milk. These miRNA exhibited sensitivity and specificity greater than 80% for differentiating between CMT+ milk and normal milk. Our findings suggest that inflammation-related miRNA expression level in the bovine milk was affected by mastitis, and miRNA in milk have potential for use as biomarkers of bovine mastitis.
组织和液体样本中的微小RNA(miRNA)已被证明与包括炎症在内的多种疾病相关。我们旨在确定牛乳炎乳汁中与炎症相关的miRNA表达水平。使用qPCR分析了患乳腺炎奶牛和正常奶牛乳汁中与炎症相关的miRNA表达水平。我们发现,在加利福尼亚乳腺炎检测阳性(CMT+)的乳汁中,miR-21、miR-146a、miR-155、miR-222和miR-383的表达水平显著上调。我们使用基于芯片的QuantStudio数字PCR系统进一步分析了这些miRNA。数字PCR结果与qPCR结果相关,表明CMT+乳汁中miR-21、miR-146a、miR-155、miR-222和miR-383上调。总之,我们鉴定出了在CMT+乳汁中上调的miRNA。这些miRNA在区分CMT+乳汁和正常乳汁方面表现出大于80%的敏感性和特异性。我们的研究结果表明,牛乳中与炎症相关的miRNA表达水平受乳腺炎影响,乳汁中的miRNA有潜力用作牛乳腺炎的生物标志物。