LRTN4RL1-AS通过miR-27a-3p靶向PPARγ调节奶牛乳腺细胞乳脂合成

doi:10.11843/j.issn.0366-6964.2023.04.011

Abstract

Abstract: This study aimed to explore the role and regulatory mechanism of lncRNA-LRTN4RL1- AS/miR-27a-3p/PPARγ axis in milk fat synthesis of dairy cows. Bovine mammary epithelial cells (BMECs) were used as experimental objects. Three biological repeats were carried out for the treatment group and the control group. Real time quantitative PCR (RT-qPCR), Western blot (WB), Oil Red O staining, triglyceride detection and double luciferase report were used to detect the effect of lncRNA-LRTN4RL1-AS on the expression of triglyceride accumulation and lipid synthesis related genes in BMECs. LRTN4RL1-AS-miR-27a-3p-PPARγ had targeting relationship of competing endogenous RNAs(ceRNA) was verified. The results showed that LRTN4RL1-AS was mainly expressed in the cytoplasm. After interfering with LRTN4RL1-AS, the lipid droplet formation and triglyceride accumulation of BMECs were significantly inhibited (P<0.05), and the expression of lipid synthesis related genes CEBPα (P<0.01), CEBPβ (P<0.01) and PPARγ were significantly down-regulated (P<0.05), and the expression of lipid catabolism gene HSL was significantly up-regulated (P<0.05). Double luciferase report assay showed that the luciferase activity in mimics-miR-27a-3p in LRTN4RL1-AS mutant group was significantly higher than that in LRTN4RL1-AS wild type group (P<0.01). It was confirmed there was a targeted binding relationship between LRTN4RL1-AS and bta-miR-27a-3p. Cotransfection experiment result showed that inhibitor-miR-27a-3p reversed the inhibition effect of interference LRTN4RL1-AS on milk fat synthesis (P<0.05), and reduced the inhibition effect of si-LRTN4RL1-AS on PPARγ gene expression. The results show that LRTN4RL1-AS regulates milk fat synthesis of BMECs by competitively binding miR-27a-3p to regulate the expression of PPARγ.

Key words: milk fat, lncRNA, PPARγ, miR-27a-3p

CLC Number:

S823.91

JIA Hongru, YANG Chaoqun, WANG Meng, WU Zhangqing, ZAN Linsen, YANG Wucai. LRTN4RL1-AS Mediates Milk Fat Synthesis in Bovine Mammary Epithelial Cells through miR-27a-3p Targeting PPARγ[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1465-1477.

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LRTN4RL1-AS Mediates Milk Fat Synthesis in Bovine Mammary Epithelial Cells through miR-27a-3p Targeting PPARγ

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