香猪睾丸间质细胞TAS1R3基因干扰后对自噬相关因子的影响

doi:10.11843/j.issn.0366-6964.2023.04.016

Abstract

Abstract: The purpose of this study was to explore the effect of TAS1R3 gene on the expression of autophagy-related genes in Congjiang Xiang pigs through mTOR. In this experiment, three 30-day-old healthy male Congjiang Xiang pigs were selected, testis tissue was collected and Leydig cells were cultured. The interference vector and empty vector were transfected into the cells, and their effects on the expression of TAS1R3 gene were detected by qRT-PCR. The effects of TAS1R3 gene interference on the expression of taste receptor family 1 receptor (TAS1R1), extracellular signal-regulated kinase 1 (ERK1), extracellular signal-regulated kinase 2 (ERK2), rapamycin target protein (mTOR), BECN1 and microtubule-associated protein 1 light chain 3 β (MAP1LC3B) were detected by qRT-PCR and Western blot. The results showed that when TAS1R3 gene was interfered, compared with shRNA-NC control group, the mRNA expression of TAS1R1, ERK1, ERK2 and mTOR genes decreased significantly(P<0.01), while the expression of autophagy factors BECN1 and MAP1LC3B increased significantly (P<0.01). The results of Western blot showed that when TAS1R3 gene was silenced, compared with shRNA-NC control group, the expression of T1R3, T1R1, p-mTOR, p-S6K1 and p-ERK12 decreased significantly(P<0.01), while the expression of autophagy protein Beclin 1 increased significantly(P<0.01). In this experiment, by transfecting TAS1R3 gene interference vector into Leydig cells, it was found that when TAS1R3 gene was interfered, it could affect the expression of mTOR gene, and then regulate the expression of downstream autophagy factors, suggesting that TAS1R3 gene is closely related to autophagy of Leydig cells, which provides a reference for further study on the regulation mechanism of TAS1R3 gene affecting the reproduction of Xiang pigs by regulating autophagy.

Key words: Congjiang Xiang pig, T1R3, RNA interference, leydig cell, autophagy

CLC Number:

S828.2

WANG Han, MENG Lijie, LIU Wenjiao, XU Yongjian, GONG Ting. Effect of TAS1R3 Gene Interference on Autophagy Related Factors in Leydig Cells of Xiang Pig[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1525-1534.

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Effect of TAS1R3 Gene Interference on Autophagy Related Factors in Leydig Cells of Xiang Pig

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