抑制miR-142-3p对奶山羊化学诱导乳腺上皮细胞泌乳功能的影响

doi:10.16431/j.cnki.1671-7236.2023.01.006

Abstract

Abstract: 【Objective】 The aim of this study was to explore the regulatory effect of miR-142-3p on the lactation function of chemical induced mammary epithelial cells (CiMECs) of dairy goats in vitro, in order to promote the development of transgenic mammary gland bioreactor and lay a foundation and provide new ideas for the commercial production of "petri dish milk".【Method】 Milk goat ear fibroblasts (GEFs) were selected as the study material.After 8 days of induction by a single inhibitor of small molecule compound TGFβR-1/ALK5 (RepSox), the GEFs were identified by morphological changes, immunofluorescence staining of specific markers and oil red O staining.After that, liposome transfection technology was used to transfect miR-142-3p inhibitor (miR-142-3p inhibitor) and inhibitor control (inhibitor-NC) respectively in CiMECs of dairy goats with successful induction, and the cells without transfection were used as blank control (BC).24 hours after transfection, the expression of miR-142-3p was detected by Real-time quantitative PCR, the cell proliferation rate was detected by CCK-8 method, the expression of β-casein was detected by Western blotting, and the content of triglyceride was determined by triglyceride enzyme method.【Result】 After 8 days of GEFs induction, insular polynucleolus-like epithelioid aggregation was formed.Immunofluorescence staining showed that the induced GEFs expressed CK14 and CK18, the specific markers of mammary epithelial cells (MECs).The results of oil red O staining showed that lipid droplets of different sizes stained red were dispersed in the cytoplasm of induced GEFs, indicating that CiMECs of dairy goats with lactation function were successfully obtained.CiMECs transfection test results of dairy goats showed that compared with blank control group, the relative expression of miR-142-3p in miR-142-3p inhibitor group was significantly decreased (P<0.05), but there was no significant difference in miR-142-3p expression in inhibitor-NC group (P>0.05).The proliferation rate, β-casein expression and triglyceride secretion of CiMECs in dairy goats with miR-142-3p inhibitor group were significantly increased (P<0.05).There were no significant differences in the proliferation rate, β-casein expression and triglyceride secretion of CiMECs in dairy goats with inhibitor-NC group (P>0.05).【Conclusion】 GEFs could be successfully transformed into dairy goat CiMECs with lactation function after 8 days of induction by RepSox.Inhibition of miR-142-3p could significantly promote the proliferation of dairy goat CiMECs and increase the secretion of β-casein and triglyceride, thus affecting the lactation function.

Key words: dairy goats; mammary epithelial cells (MECs); miR-142-3p

CLC Number:

S827

PAN Siyao, ZHANG Dandan, LIU Quanhui, WANG Guodong, LYU Danwei, HUANG Ben. Effects of Inhibition of miR-142-3p on Lactation Function of Chemical Induced Mammary Epithelial Cells in Dairy Goats[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 58-66.

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Effects of Inhibition of miR-142-3p on Lactation Function of Chemical Induced Mammary Epithelial Cells in Dairy Goats

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