猪子宫腔液外泌体来源的TIMP2蛋白对胚胎附植的影响

doi:10.11843/j.issn.0366-6964.2022.04.012

Abstract

Abstract: This study aimed to investigate the effect of TIMP2 protein derived from porcine uterine fluid exosomes on embryo implantation during early pregnancy. Exosomes were isolated from the uterine fluid of Yorkshire pigs at day 9 (n=3) and 12 (n=3) of pregnancy by ultracentrifugation. The relative abundance of TIMP2 protein in exosomes was determined using Western blot (WB)， and the expression and location of TIMP2 in uterus was detected by immunohistochemistry. Subsequently, co-culture experiments of exosomes and porcine trophoblast cells (PTr2) were used to investigate the targeted transport of exosomes for TIMP2 protein. Finally, the TIMP2 gene was interfered or overexpressed in PTr2, and the effects of TIMP2 on proliferation and migration of PTr2 cells were determined using CCK-8, EdU and wound healing assay. The exosomes were successfully isolated from uterine fluid by ultracentrifugation, and their particle sizes were mainly distributed from 30 to 150 nm. WB results showed that the TIMP2 protein abundance in exosomes of pig uterine fluid at day 12 of pregnancy was significantly increased (P<0.05). While the co-culture experiments of exosomes and trophoblast cells showed that TIMP2 protein could be targeted transport to trophoblast cells through exosomes. Immunohistochemical results showed that TIMP2 protein was found in both the embryo and endometrium, and the abundance level in endometrium at day 12 of pregnancy was significantly higher than that at day 9 of pregnancy (P<0.01). In addition, in vitro cell culture experiments found that interfering with TIMP2 gene in PTr2 significantly enhanced cell proliferation (P<0.01) and migration ability (P<0.05)， while the overexpression of TIMP2 impaired the proliferation (P<0.01) and migration ability (P<0.05). These results suggest that TIMP2 protein in uterine fluid exosomes may be secreted by the maternal endometrium, and could be targeted into the trophoblast cells through exosomes to regulate their proliferation and migration, thereby affecting embryo implantation.

Key words: exosomes, TIMP2, PTr2, embryo implantation

CLC Number:

S828.3

KUANG Jingjing, HE Yanjuan, HU Qun, GU Ting, WU Zhenfang, CAI Gengyuan, HONG Linjun. Effect of TIMP2 Protein Derived from Porcine Uterine Fluid Exosomes on Embryo Implantation During Early Pregnancy[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1122-1132.

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Effect of TIMP2 Protein Derived from Porcine Uterine Fluid Exosomes on Embryo Implantation During Early Pregnancy

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