Mechanism of PBMCs-derived exosomes on improving endometrial receptivity by miR-1306
10.3760/cma.j.cn101441-20220620-00267
- VernacularTitle:PBMCs源性外泌体经miR-1306改善子宫内膜容受性的机制研究
- Author:
Yungai XIANG
1
;
Guo YU
1
;
Yan LI
1
;
Lijing WAN
1
;
Liying MA
1
;
Xujing GENG
1
;
Linlin TAN
1
;
Li TAN
1
Author Information
1. 郑州大学第二附属医院生殖医学部，郑州 450001
- Publication Type:Journal Article
- Keywords:
Exosomes;
Implantation disorder;
Peripheral blood mononuclear cell;
miR-1306;
Endometrial receptivity
- From:
Chinese Journal of Reproduction and Contraception
2023;43(4):381-388
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the mechanism of peripheral blood mononuclear cells (PBMCs)-derived exosomes on improving endometrial receptivity by miR-1306.Methods:The models of mice with implantation disorders were prepared by subcutaneous injection of mifepristone. And they were divided into implantation disorder group and PBMCs intervention group, 12 mice in each group. The normal group was set up. PBMCs intervention group was given intrauterine injection of PBMCs-derived exosomes, while normal group and implantation disorder group were given the same volume of buffer solution. The pregnancy rates and number of embryos implantation in different groups were compared. The expression of miR-1306 in endometrial tissues was detected by RT-PCR. The levels of reactive oxygen species (ROS), interleukin-6 (IL-6), monocyte chemcattractant protein-1 (MCP-1) and superoxide dismutase (SOD) in endometrial tissues were detected by enzyme-linked immunosorbent assay. The expressions of pregnancy-related proteins in endometrial tissues were detected by Western blotting. The endometrial epithelial cells were divided into control group, experimental group, negative control group and miR-1306 inhibitor group. Except control group, cells were co-cultured with PBMCs-derived exosomes by Transwell in the other groups. The activity, proliferation and apoptosis of cells were detected by MTT, Edu and Annexin-V/PI flow, respectively. The level of ROS in cells was detected by kits. And expressions of the related proteins were detected by Western blotting.Results:The pregnancy rate [8.33% (1/12), number of embryos implantation [0(0, 0)], expression of miR-1306 in endometrial tissue (0.24±0.05) and SOD level [(5.66±0.72) U/mL] in implantation disorder group were significantly lower than those in normal group [100% (12/12), 16.50 (14.00, 19.00), 1.03±0.05, (8.69±1.21) U/mL, P<0.05], while levels of ROS [(4.87±0.39) U/mL], IL-6 [(116.51±5.78) ng/L] and MCP-1 in endometrial tissues [(36.84±3.56) μg/L], and expressions of KIR2DL4 (0.87±0.06), nuclear factor erythroid-2-related factor 2 (Nrf2, 0.76±0.06), Kelch-like ECH-associated protein1 (Keap1, 0.79±0.05), receptor interacting protein 1 (RIP1, 0.94±0.04) and RIP3 (0.86±0.05) were significantly higher than those in normal group [(2.41±0.19) U/mL, (83.79±6.68) ng/L, (12.32±2.09) ng/mL, 0.27±0.03, 0.31±0.05, 0.23±0.04, 0.34±0.03, 0.31±0.05, all P<0.05]. The pregnancy rate [75.00% (9/12)], number of embryos implantation [13.00 (13.00, 14.75)], expression of miR-1306 in endometrial tissues (0.82±0.05) and SOD level [(7.24±0.84) U/mL] in PBMCs intervention group were significantly higher than those in implantation disorder group (all P<0.05), while levels of ROS [(3.43±0.30) U/mL], IL-6 [(94.69±3.99) ng/L] and MCP-1 in endometrial tissues [(27.03±3.48) μg/L], and expressions of KIR2DL4 (0.54±0.08), Nrf2 (0.48±0.05), Keap1 (0.43±0.05), RIP1 (0.56±0.05) and RIP3 (0.49±0.03) were significantly lower than those in implantation disorder group (all P<0.05). The cells activity [(126.63±1.25)%], proliferation rate [(53.54±2.82)%] and ROS level [(3.12±0.31) U/mL] in experimental group were significantly higher than those in control group [100%, (23.18±3.07)%, (2.51±0.28) U/mL, all P<0.05], while apoptosis rate [(5.69±0.47)%], expressions of KIR2DL4 (0.36±0.06), Nrf2 (0.30±0.06), Keap1 (0.26±0.04), RIP1 (0.27±0.05) and RIP3 (0.26±0.06) were lower than those in control group [(27.13±2.97)%, 0.84±0.06, 0.75±0.05, 0.68±0.05, 0.80±0.06, 0.80±0.07, all P<0.05]. The cells activity [(83.48±5.34)%] and proliferation rate [(38.42±4.28)%] in miR-1306 inhibitor group were significantly lower than those in negative control group [(127.12±4.08)%, (53.57±2.09)%, all P<0.05], while apoptosis rate [(13.63±1.77)%], ROS level [(6.49±0.62) U/mL], expressions of KIR2DL4 (0.67±0.07), Nrf2 (0.57±0.05), Keap1 (0.50±0.05), RIP1 (0.64±0.06) and RIP3 (0.61±0.08) were significantly higher than those in negative control group [(5.71±0.78)%, (3.23±0.31) U/mL, 0.36±0.07, 0.30±0.07, 0.27±0.06, 0.28±0.07, 0.28±0.06, all P<0.05]. Conclusion:PBMCs-derived exosomes can improve pregnancy rate in mice with implantation disorders, which may improve inflammatory response by promoting miR-1306 expression and inhibiting KIR2DL4 expression, and is also related to relieving excessive oxidative stress and apoptosis of endometrium.
