基于PI3K/Akt通路探究蒙古羊ADAMTS1基因的作用机制

doi:10.16431/j.cnki.1671-7236.2023.09.002

Abstract

Abstract: 【Objective】 This study was aimed to analyze the expression pattern of a disintegrin and metalloproteinase with thrombospondin motifs 1 (ADAMTS1) on PI3K/Akt pathway in ovary of twin and singleton Mongolian sheep at different stages of estrous cycle,and explore the mechanism of ADAMTS1 gene influencing reproductive traits in Mongolian sheep.【Method】 A total of 40 Mongolian sheep with twin and single births were selected for natural estrus,ewe estrus and diestrus periods were determined.Groups of 10 Mongolian sheep each in twin estrus,twin diestrus,single estrus and single diestrus groups. The granulosa cells of 2-3 years old Mongolian sheep were extracted by syringe,which were divided into control and experiment groups.No drug added to granulosa cells in control group,and 15 μmol/L LY294002 added to granulosa cells in experiment group,the cell viability were measured at 24,48 and 72 h.The relative expression of ADAMTS1,PI3K,PTEN,Akt,RPS6,Bcl-2 and BAX genes in ovary and granulosa cells of ewe were detected by Real-time quantitative PCR.The expression of ADAMTS1,PI3K and Akt proteins in ovary and granulosa cells were detected by Western blotting.【Result】 The results of Real-time quantitative PCR showed that the expression of ADAMTS1 and Bcl-2 genes in twin estrus group were significantly higher than that in other groups (P＜0.05).The expression of PI3K,Akt and PTEN genes in twin estrus and twin diestrus groups were significantly higher than that in single estrus and single diestrus groups (P＜0.05).The expression of RPS6 gene in single diestrus group was significantly lower than that in other groups (P＜0.05).Compared with estrus,the expression of BAX gene was significantly upregulated during diestrus (P<0.05).Western blotting results showed that the expression of ADAMST1 protein in twin estrus group was significantly higher than that in other groups (P＜0.05).Compared with single group,the expression of PI3K and Akt proteins in twin group were upregulated,the expression of PI3K protein in twin estrus and twin diestrus were significantly higher than that in single estrus and single diestrus groups (P＜0.05),and the expression of Akt protein in twin estrous was the highest,which were significantly higher than that in other groups (P＜0.05).Compared with control group,the granulosa cells in experiment group of Mongolian sheep had abnormal morphology and slow proliferation,and the relative expression of ADAMTS1, PI3K and Akt genes and proteins were significantly decreased (P＜0.05).【Conclusion】 The expression trends of ADAMTS1,PI3K and Akt genes and proteins were consistent and showed a positive correlation in ovary and granulosa cells of Mongolian sheep,suggested that ADAMTS1 gene played an important role in twin birth of Mongolian sheep mediated by PI3K/Akt signaling pathway.

Key words: Mongolian sheep; ADAMTS1 gene; PI3K/Akt pathway; twin; single; estrus cycle

CLC Number:

S814.8

CHEN Xiaoliang, ZHAO Mengyao, LI Xiunan, SU Lede, TIAN Ying, JIAN Ruizhen, BAYIN Jiri Gala, YANG Yanyan. Study on the Mechanism of ADAMTS1 Gene in Mongolian Sheep Based on PI3K/Akt Pathway[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3459-3468.

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Study on the Mechanism of ADAMTS1 Gene in Mongolian Sheep Based on PI3K/Akt Pathway

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