文献类型： 外文期刊

作者： Gao, Ruonan ¹ ; Li, Qingchun ¹ ; Qiu, Meiyu ¹ ; Xie, Su ¹ ; Sun, Xiaomei ¹ ; Huang, Tao ¹ ;

作者机构： 1.Shihezi Univ, Coll Anim Sci & Technol, 221 North Fourth Rd, Shihezi 832000, Peoples R China

2.Xinjiang Acad Anim Sci, Inst Biotechnol, Urumqi 830000, Peoples R China

3.Huazhong Agr Univ, Coll Anim Sci & Technol, Key Lab Anim Breeding & Reprod, Minstry Educ, Wuhan 430070, Peoples R China

4.Xinjiang Husb S&T Co Ltd, Xinjiang Pig Breeding Engn Technol Res Ctr, Changji 831100, Peoples R China

关键词： Biomarkers; Early Pregnancy; Exosomal miRNAs; ITGA4; miR-192

期刊名称：ANIMAL BIOSCIENCE （ 影响因子：2.2； 五年影响因子：2.3 ）

ISSN： 2765-0189

年卷期： 2023 年 36 卷 9 期

页码：

收录情况： SCI

摘要： Objective: The study was conducted to screen differentially expressed miRNAs in sows at early pregnancy by high-throughput sequencing and explore its mechanism of action on embryo implantation. Methods: The blood serum of pregnant and non-pregnant LandracexYorkshire sows were collected 14 days after artificial insemination, and exosomal miRNAs were purified for high throughput miRNA sequencing. The expression patterns of 10 differentially expressed (DE) miRNAs were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The qRT-PCR quantified the abundance of serum exosomal miR-192 in pregnant and control sows, and the diagnostic power was assessed by receiver operating characteristic (ROC) analysis. The target genes of DE miRNAs were predicted with bioinformatics software, and the functional and pathway enrichment analysis was performed on gene ontology and the Kyoto encyclopedia of genes and genomes terms. Furthermore, a luciferase reporter system was used to identify the target relation between miR-192 and integrin alpha 4 (ITGA4), a gene influencing embryo implantation in pigs. Finally, the expression levels of miRNAs and the target gene ITGA4 were analyzed by qRT-PCR, and western blot, with the proliferation of BeWo cells detected by cell counting kit-8 (CCK-8).Results: A total of 221 known miRNAs were detected in the libraries of the pregnant and non-pregnant sows, of which 55 were up-regulated and 67 were down-regulated in the pregnant individuals compared with the non-pregnant controls. From these, the expression patterns of 10 DE miRNAs were validated. The qRT-PCR analysis further confirmed a significantly higher expression of miR-192 in the serum exosomes extracted from pregnant sows, when compared to controls. The ROC analysis revealed that miR-192 provided excellent diagnostic accuracy for pregnancy (area under the ROC curve [AUC] = 0.843; p>0.001). The dual-luciferase reporter assay indicated that miR-192 directly targeted ITGA4. The protein expression of ITGA4 was reduced in cells that overexpressed miR-192. Overexpression of miR-192 resulted in the decreased proliferation of BeWo cells and regulated the expression of cell cycle-related genes.Conclusion: Serum exosomal miR-192 could serve as a potential biomarker for early pregnancy in pigs. miR-192 targeted ITGA4 gene directly, and miR-192 can regulate cellular proliferation.