The serine protease 2 gene regulates lipid metabolism through the LEP/ampkα1/SREBP1 pathway in bovine mammary epithelial cells.
The serine protease 2 gene regulates lipid metabolism through the LEP/ampkα1/SREBP1 pathway in bovine mammary epithelial cells.
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DOI：
10.1016/j.bbrc.2024.149558
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发表时间：
2024-01
影响因子：
3.1
通讯作者：
Huixian Lu;Zhihui Zhao;Haibin Yu;Ambreen Iqbal;P. Jiang
中科院分区：
文献类型：
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作者：
Huixian Lu;Zhihui Zhao;Haibin Yu;Ambreen Iqbal;P. Jiang
分子育种在21世纪给牛奶市场和生产系统带来了重大变革。奶制品生产的主要经济特性与乳脂含量有关。我们先前的转录组分析表明，丝氨酸蛋白酶2（PRSS 2）是一个候选基因，可能会影响中国荷斯坦奶牛乳腺上皮细胞（BMEC）的乳脂合成。为了阐明PRSS 2基因在乳脂合成中的功能，我们构建了PRSS 2过表达和干扰的载体，并评估了BMEC中细胞内甘油三酯（TG）、胆固醇（CHOL）和非酯化脂肪酸（NEFA）的含量。采用气相色谱-质谱联用（GC-MS）技术对脂肪酸种类和组分进行了定量分析。通过qPCR、ELISA和WB技术验证了PRSS 2介导的调控途径。基于我们的研究结果，PRSS 2成为调节相关基因表达的关键基因，从而通过瘦素（LEP）/腺苷酸活化蛋白激酶，α 1催化亚基（AMPKα1）/固醇调节元件结合蛋白1（SREBP 1）途径通过抑制TG和CHOL积累而对脂质代谢做出实质性贡献，同时潜在地促进NEFA在BMEC中的合成。此外，PRSS 2基因通过调节与LEP/AMPKα1/SREBP 1通路相关的基因增强细胞内中链和长链脂肪酸代谢，导致不饱和脂肪酸C17：1 N7和C22：4 N6含量增加。本研究为进一步研究PRSS 2影响奶牛脂质代谢的分子机制提供了一个强有力的理论框架。
Molecular breeding has brought about significant transformations in the milk market and production system during the twenty-first century. The primary economic characteristic of dairy production pertains to milk fat content. Our previous transcriptome analyses revealed that serine protease 2 (PRSS2) is a candidate gene that could impact milk fat synthesis in bovine mammary epithelial cells (BMECs) of Chinese Holstein dairy cows. To elucidate the function of thePRSS2gene in milk fat synthesis, we constructed vectors forPRSS2overexpression and interference and assessed intracellular triglycerides (TGs), cholesterol (CHOL), and nonesterified fatty acid (NEFA) contents in BMECs. Fatty acid varieties and components were also quantified using gas chromatography‒mass spectrometry (GC‒MS) technology. The regulatory pathway mediated byPRSS2was validated through qPCR, ELISA, and WB techniques. Based on our research findings,PRSS2emerges as a pivotal gene that regulates the expression of associated genes, thereby making a substantial contribution to lipid metabolism via the leptin (LEP)/Adenylate-activated protein kinase, alpha 1 catalytic subunit (AMPKα1)/sterol regulatory element binding protein 1(SREBP1)pathway by inhibiting TGs and CHOL accumulation while potentially promoting NEFA synthesis in BMECs. Furthermore, thePRSS2gene enhances intracellular medium- and long-chain fatty acid metabolism by modulating genes related to theLEP/AMPKα1/SREBP1pathway, leading to increased contents of unsaturated fatty acids C17:1N7 and C22:4N6. This study provides a robust theoretical framework for further investigation into the underlying molecular mechanisms through whichPRSS2influences lipid metabolism in dairy cows.