Transcriptomic Analysis of Short/Branched-Chain Acyl-Coenzyme a Dehydrogenase Knocked Out bMECs Revealed Its Regulatory Effect on Lipid Metabolism.
Transcriptomic Analysis of Short/Branched-Chain Acyl-Coenzyme a Dehydrogenase Knocked Out bMECs Revealed Its Regulatory Effect on Lipid Metabolism.
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敲除 bMEC 的短/支链酰基辅酶 a 脱氢酶的转录组分析揭示了其对脂质代谢的调节作用
DOI：
10.3389/fvets.2021.744287
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发表时间：
2021
影响因子：
3.2
通讯作者：
Zhao Z
中科院分区：
文献类型：
--
作者：
Jiang P;Iqbal A;Wang M;Li X;Fang X;Yu H;Zhao Z
酶的酶脱氢酶家族包括短/分支链酰基-COA脱氢酶（ACADSB），催化脂肪酸代谢中的脂肪酸代谢中的酰基-COA衍生物的脱氢作用，这表明我们先前的发现是通过对牛奶脂肪的重要群体进行比较的牛奶症状。奶牛生产高脂和在本研究中，低脂牛奶以及基因功能验证研究。和6,098 GO术语丰富了，其中637个GO术语大大增强，例如磷脂 - 转移ATPase Atpase活性（GO：0004012），脂蛋白脂肪酶活性（GO：0004465），酰基-COA拼命的活性（GO：0016215）。脂肪酸的废奴（路径： 01212），甘油的代谢（路径：00561）和脂肪细胞的信号传导（路径：04920）。当Acads B基因揭示了rt2 prifiler pcr rane的损失时，BMEC中的CHOL，TGS和FFA含量降低了。 ，包括ACADL，ACOX2，ACAT2和FABP3。总之，目前的发现表明，ACADSB是BMEC中脂质代谢的关键调节剂，这也是对与脂质和脂肪酸代谢相关的基因功能的未来研究的有用遗传材料和工具。
The acyl-CoA dehydrogenase family of enzymes includes short/branched-chain acyl-CoA dehydrogenase (ACADSB), which catalyzes the dehydrogenation of acyl-CoA derivatives in fatty acid metabolism. Our previous findings suggested that ACADSB was a critical candidate gene affecting milk fat synthesis by comparing the transcriptome in bovine mammary epithelial cells (bMECs) from Chinese Holstein dairy cows producing high-fat and low-fat milk as well as gene functional validation studies on the cellular level. In the present study, ACADSB in bMECs was knocked out (KO) using a CRISPR/Cas9 system, and mRNA transcriptome was further sequenced to verify the function of the ACADSB gene and analyze its correlation with lipid metabolism. The findings revealed that 15,693 genes were expressed, 1,548 genes were differentially expressed genes (DEGs), and 6,098 GO terms were enriched, of which 637 GO terms were greatly enhanced, such as phospholipid-translocation ATPase activity (GO:0004012), lipoprotein lipase activity (GO:0004465), acyl-CoA desaturase activity (GO:0016215), and so on. The analysis by KEGG showed that DEGs were distributed over 247 pathogens, of which 49 were significantly enriched, including the metabolism of fatty acids (PATH: 01212), metabolism of glycerolipid (PATH: 00561), and signaling of adipocytokines (PATH: 04920). The CHOL, TGs and FFA contents in bMECs were reduced when the ACADSB gene was knocked out. The RT2 Profiler PCR array also revealed that the loss of the ACADSB gene changed the expression levels of functional genes involved in lipid metabolism, including ACADL, ACOX2, ACAT2, and FABP3. In conclusion, the current findings show that ACADSB is a key regulator of lipid metabolism in bMECs. The ACADSB−/− bMECs could also be useful genetic material and tools for future research into gene functions related to lipid and fatty acid metabolism. It will be valuable for revealing the gene regulatory roles and molecular mechanisms in milk fat synthesis.
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