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蛋白质组学对牛乳腺上皮细胞中9,11-共轭亚油酸生物合成基因网络的见解

Proteomics Insights into the Gene Network of 9, 11-Conjugated Linoleic Acid Biosynthesis in Bovine Mammary Gland Epithelial Cells.

作者信息

Peng Liying, Bai Ge, Wang Chunzheng, Dong Jianan, Liu Yongjun, Sun Zhe, Zhen Yuguo, Qin Guixin, Zhang Xuefeng, Demelash Natnael, Wang Tao

机构信息

JLAU-Borui Dairy Science and Technology R&D Center, Key Laboratory of Animal Nutrition and Feed Science of Jilin Province, Key Laboratory of Animal Production Product Quality and Security Ministry of Education, College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China.

Changchun Institute of Biological Products Co., Ltd., Changchun 130012, China.

出版信息

Animals (Basel). 2022 Jul 2;12(13):1718. doi: 10.3390/ani12131718.

DOI:10.3390/ani12131718
PMID:35804617
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9264836/
Abstract

The objective of the study was to elucidate the stearoyl-coenzyme A desaturase (SCD1)-dependent gene network of c9, t11-CLA biosynthesis in MAC-T cells from an energy metabolism perspective. The cells were divided into the CAY group (firstly incubated with CAY10566, a chemical inhibitor of SCD1, then incubated with trans-11-octadecenoic acid, (TVA)), the TVA group (only TVA), and the control group (without CAY, TVA). The c9, t11-CLA, and TVA contents were determined by gas chromatography. The mRNA levels of SCD1 and candidate genes were analyzed via real-time PCR. Tandem mass tag (TMT)-based quantitative proteomics, bioinformatic analysis, parallel reaction monitoring (PRM), and small RNA interference were used to explore genes involved in the SCD1-dependent c9, t11-CLA biosynthesis. The results showed that the SCD1 deficiency led by CAY10566 blocked the biosynthesis of c9, t11-CLA. In total, 60 SCD1-related proteins mainly involved in energy metabolism pathways were primarily screened by TMT-based quantitative proteomics analysis. Moreover, 17 proteins were validated using PRM analysis. Then, 11 genes were verified to have negative relationships with SCD1 after the small RNA interference analysis. Based on the above results, we concluded that genes involved in energy metabolism pathways have an impact on the SCD1-dependent molecular mechanism of c9, t11-CLA biosynthesis.

摘要

本研究的目的是从能量代谢角度阐明MAC-T细胞中硬脂酰辅酶A去饱和酶1（SCD1）依赖性的c9，t11-共轭亚油酸（CLA）生物合成基因网络。将细胞分为CAY组（先用SCD1的化学抑制剂CAY10566孵育，再用反式-11-十八碳烯酸（TVA）孵育）、TVA组（仅用TVA）和对照组（不添加CAY、TVA）。采用气相色谱法测定c9，t11-CLA和TVA的含量。通过实时荧光定量PCR分析SCD1及候选基因的mRNA水平。运用基于串联质谱标签（TMT）的定量蛋白质组学、生物信息学分析、平行反应监测（PRM）和小干扰RNA来探索参与SCD1依赖性c9，t11-CLA生物合成的基因。结果显示，CAY10566导致的SCD1缺乏阻断了c9，t11-CLA的生物合成。基于TMT的定量蛋白质组学分析初步筛选出总共60种主要参与能量代谢途径的SCD1相关蛋白。此外，通过PRM分析验证了17种蛋白。小干扰RNA分析后，证实有11个基因与SCD1呈负相关。基于上述结果，我们得出结论，参与能量代谢途径的基因对SCD1依赖性的c9，t11-CLA生物合成分子机制有影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ab/9264836/8664e5c68599/animals-12-01718-g010.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ab/9264836/25750f31021f/animals-12-01718-g008.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ab/9264836/8664e5c68599/animals-12-01718-g010.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ab/9264836/9f8bf5037801/animals-12-01718-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ab/9264836/e3101a26fe61/animals-12-01718-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/76ab/9264836/25750f31021f/animals-12-01718-g008.jpg
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