Han Xuelei, Qi Kunlong, Song Chenglei, Dou Yaqing, Liu Yingke, Li Chenlei, Wei Yilin, Qiao Ruimin, Li Xiuling, Yang Feng, Wang Kejun, Li Xinjian
College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China.
Life (Basel). 2022 Nov 7;12(11):1814. doi: 10.3390/life12111814.
A miRNA-mRNA combination analysis was performed on the longissimus dorsi muscle of adult Queshan Black and Large White pigs by RNA-seq technology to reveal the molecular mechanism affecting pork quality traits. The sequencing results showed that 39 miRNAs were differentially expressed between Queshan Black and Large White pigs, which targeted 5234 mRNAs, and 15 differentially expressed miRNAs targeted 86 differentially expressed mRNAs. The qRT-PCR results showed that miRNAs showed similar expression patterns to RNA-seq. The GO analysis indicated that differentially expressed miRNAs with differential target mRNAs were primarily involved in biological processes such as phospholipase activity, MAP-kinase scaffold activity, lipase activity, and regulation of the extent of cell growth. The KEGG analysis also revealed that such mRNAs were significantly enriched in the ECM-receptor interaction, sphingolipid metabolism, apoptosis, PI3K-Akt signaling pathway, and AMPK signaling pathway. In addition, software predictions showed that 17 (13 of which were upregulated and four were downregulated) of 39 differentially expressed miRNAs targeted 118 negatively correlated expression mRNAs. The upregulated miRNAs contained 103 negatively correlated target mRNAs, whereas the downregulated miRNAs contained 15 negatively correlated target mRNAs. The GO analysis showed that such mRNAs were primarily involved in MAP-kinase scaffold activity, myoblast development, and peptidyl-lysine methylation, and the KEGG analysis showed significant enrichment in ECM-receptor interaction and focal adhesion. The functional enrichment analysis of miRNA target genes revealed that miR-328 was screened out as a key miRNA, and preliminary functional validation was performed. Moreover, the overexpressed miR-328 could affect the expression of proliferation-related genes, such as CDK2, CDK4, CCNB1, CCND1, CCNE1, and PCNA. These results indicated that miR-328 may regulate fat deposition and affect meat quality by influencing related pathways. This study revealed that the miRNA-mRNA regulatory axis affects fat deposition and skeletal muscle development, which provides a theoretical basis for further study on the molecular mechanism of meat quality.
采用RNA测序技术对成年确山黑猪和大白猪的背最长肌进行miRNA-mRNA联合分析，以揭示影响猪肉品质性状的分子机制。测序结果表明，确山黑猪和大白猪之间有39个miRNA差异表达，这些miRNA靶向5234个mRNA，其中15个差异表达的miRNA靶向86个差异表达的mRNA。qRT-PCR结果显示，miRNA的表达模式与RNA测序相似。GO分析表明，具有差异靶mRNA的差异表达miRNA主要参与磷脂酶活性、MAP激酶支架活性、脂肪酶活性和细胞生长程度调节等生物学过程。KEGG分析还显示，这些mRNA在细胞外基质-受体相互作用、鞘脂代谢、细胞凋亡、PI3K-Akt信号通路和AMPK信号通路中显著富集。此外，软件预测显示，39个差异表达的miRNA中有17个（其中13个上调，4个下调）靶向118个负相关表达的mRNA。上调的miRNA包含103个负相关的靶mRNA，而下调的miRNA包含15个负相关的靶mRNA。GO分析表明，这些mRNA主要参与MAP激酶支架活性、成肌细胞发育和肽基赖氨酸甲基化，KEGG分析显示在细胞外基质-受体相互作用和粘着斑中显著富集。miRNA靶基因的功能富集分析筛选出miR-328作为关键miRNA，并进行了初步功能验证。此外，过表达的miR-328可影响增殖相关基因如CDK2、CDK4、CCNB1、CCND1、CCNE1和PCNA的表达。这些结果表明，miR-328可能通过影响相关途径调节脂肪沉积并影响肉质。本研究揭示了miRNA-mRNA调控轴影响脂肪沉积和骨骼肌发育，为进一步研究肉质分子机制提供了理论依据。
