miR-455-3p对奶山羊化学诱导乳腺上皮细胞乳脂代谢的影响

doi:10.16431/j.cnki.1671-7236.2024.05.018

Abstract

Abstract: 【Objective】 This study was aimed to explore the effect of miR-455-3p on milk fat metabolism in chemically induced mammary epithelial cells (CiMECs) of dairy goats,so as to provide a theoretical basis for the study of microRNA (miRNA) regulation of mammary gland function in goats.【Method】 Goat fibroblasts (GEFs) were induced by a single small molecule compound RepSox (TGFβR-1/ALK5 inhibitor),and the morphological changes of the cells were observed after 8 days of induction,and immunofluorescence staining and oil red O staining,which were specific markers,were used to identify whether the mammary epithelial cells were successfully induced.The overexpression vectors (miR-455-3p mimics and miR-455-3p mimics-NC) and interference vectors (miR-455-3p inhibitors and miR-455-3p inhibitors-NC) for miR-455-3p were transfected into CiMECs using liposomes,which were analyzed by the cell proliferation and toxicity assay (CCK-8) kit,triglyceride assay kit and Real-time quantitative PCR to detect the proliferation rate,triglyceride secretion and the expression of milk fat metabolism genes of miR-455-3p for CiMECs.【Result】 GEFs induced the formation of insular,multinucleated and cobblestone cell shapes similar to goat mammary epithelial cells after 8 days of induction.The immunofluorescence results showed that CiMECs could express epithelial-specific marker antigens,including cytokeratin 14 (CK14) and CK18.The results of oil-red O staining showed that CiMECs possessed the function of secretion of lipid droplets.The CiMECs transfection results showed that compared with control group,miR-455-3p mimics extremely significantly promoted the expression of miR-455-3p (P＜0.01),and miR-455-3p inhibitors significantly inhibited the expression of miR-455-3p expression (P＜0.05).The results of CCK-8 and triglyceride assay showed that compared with control group,the cell proliferation rate and triglyceride secretion were extremely significantly decreased in miR-455-3p mimics group (P＜0.01),and the cell proliferation rate and triglyceride secretion were significantly increased in miR-455-3p inhibitors group (P＜0.05).Real-time quantitative PCR results showed that compared with control group,the expression of HADHB gene was extremely significantly decreased in CiMECs after induction (P＜0.01),was extremely significantly increased in miR-455-3p mimics group (P＜0.01),and was significantly decreased in miR-455-3p inhibitors group (P＜0.05).【Conclusion】 The inhibition of miR-455-3p promoted CiMECs proliferation and reduced the expression of the fat metabolism gene HADHB in dairy goats,thereby increasing the fatty acid content of goat milk.The results could provide a basis for the subsequent use of miRNA to improve the milk quality in goats.

Key words: dairy goats; chemically induced mammary epithelial cells (CiMECs); miR-455-3p; milk fat

CLC Number:

S814.8

LI Weiqing, WANG Guodong, LYU Danwei, ZHANG Dandan, LIU Quanhui, PAN Siyao, HUANG Ben. Effect of miR-455-3p on Milk Fat Metabolism in Chemically Induced Mammary Epithelial Cells in Dairy Goats[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(5): 1969-1978.

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Effect of miR-455-3p on Milk Fat Metabolism in Chemically Induced Mammary Epithelial Cells in Dairy Goats

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