Document Type : Research Paper
Author
Department of Animal Sciences, Faculty of Agriculture, Yasouj University
Abstract
Candidate miRNAs were selected based on a multi-step criterion. Initially, a systematic review of previous studies that experimentally (via RNA-Seq, qPCR, or DNA microarray) reported the specific expression of miRNAs in inflammatory or infectious mastitis samples in cattle was conducted. Subsequently, miRNAs identified in at least two independent studies or in more than one reliable database (miRBase, NCBI GEO, miRTarBase) as being involved in inflammatory or immune responses were considered. Using bioinformatics analyses and reputable databases such as MirtarBase, TargetScan, DAVID, and NCBI, candidate genes associated with mastitis were identified, and their target signaling pathways were examined. This multi-step selection process was employed to ensure the accuracy and reliability of the selected miRNAs. The results of these analyses identified five key miRNAs, including bta-mir-146a, bta-mir-16a, bta-mir-181, bta-mir-21-5p, and bta-mir-223, each of which plays a role in critical biological pathways such as TLR4/NF-κB, MAPK, PI3K-AKT, TGFβ, and estrogen and progesterone hormone signaling. Furthermore, the findings revealed that bta-mir-146a, bta-mir-21-5p, and bta-mir-223 influence 7, 9, and 11 key proteins, respectively, within these biological pathways. These pathways are involved in regulating immune response, inflammation, cell growth, differentiation, and apoptosis, disruptions of which may lead to the onset or exacerbation of mastitis. Each of these miRNAs, by targeting multiple key proteins, could serve as sensitive and specific biomarkers for the early detection of mastitis in dairy cattle.
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