【摘要】 为了解双峰驼血浆外泌体对其脂质代谢的调控机制，试验分别选取体型消瘦和正常双峰驼各5头，采血后对血浆外泌体进行了提取和检测，提取外泌体总RNA后进行miRNAs测序与差异表达miRNAs的筛选，采用实时荧光定量PCR法对不同体型双峰驼血浆外泌体的差异表达miRNAs进行验证，并利用在线软件对筛选到的miRNAs靶基因进行预测、GO功能和KEGG通路富集分析。结果表明:体型消瘦和正常双峰驼血浆外泌体均呈现杯状或圆形，平均径粒分别为123.6 nm和117.9 nm，分布范围为50～200 nm;与体型正常双峰驼相比，体型消瘦双峰驼血浆外泌体中miR-21-5p、miR-1246、miR-320c等15个miRNAs下调表达，miR-144-5p、miR-4454L-2、miR-582-5p等26个miRNAs上调表达，其中miR-144-5p与脂质代谢密切相关; miR-144-5p表达水平在不同体型间的变化趋势与测序结果一致;在预测到的miR-144-5p靶基因中，PGC-1α基因与脂质代谢密切相关; PGC-1α基因参与的与脂质代谢相关的GO功能二级条目包括胆固醇平衡、脂质磷酸化，KEGG信号通路包括三酰甘油代谢、胆固醇代谢、脂质与动脉粥样硬化。说明双峰驼血浆外泌体miR-144-5p可能通过调节PGC-1α基因参与脂质代谢过程。更多还原

【Abstract】 To understand the regulatory mechanism of plasma exosomes on lipid metabolism in bactrian camels,5 emaciated and 5 normal bactrian camels were selected respectively in this experiment. After blood collection,the plasma exosomes were extracted and detected. After extracting the total RNA of the exosomes,miRNAs sequencing and screening of differentially expressed miRNAs were performed. The differentially expressed miRNAs in exosomes of bactrian camels of different body types were verified by real-time fluorescence quantitative PCR.The target genes of the screened miRNAs were predicted using online software,and GO functions and KEGG pathway enrichment analysis were conducted. The results showed that the plasma exosomes of both emaciated and normal bactrian camels were cup-shaped or round,with average particle sizes of 123. 6 nm and 117. 9 nm respectively,and the distribution range was 50-200 nm. Compared with normal-sized bactrian camels,15 miRNAs such as miR-21-5p、miR-1246 and miR-320c were down-regulated in the plasma exosomes of emaciated bactrian camels,and 26 miRNAs such as miR-144-5p、miR-4454 and miR-582-5p were up-regulated. Among them,miR-144-5p was closely related to lipid metabolism.The changing trend of miR-144-5p gene expression level analyzed by real-time fluorescence quantitative PCR among different body types was consistent with the sequencing results. Among the predicted miR-144-5p target genes,the PGC-1α gene was closely related to lipid metabolism. The GO functional secondary items related to lipid metabolism involving the PGC-1α gene included cholesterol balance and lipid phosphorylation. The KEGG signaling pathways included glycerolipid metabolism,cholesterol metabolism,lipids and atherosclerosis. The results indicated that miR-144-5p in Bactrian camel plasma exosomes might be involved in the lipid metabolism process by regulating the PGC-1ɑ gene.更多还原