Effects of Interleukin-1β, -1Rα on the Expression of Matrix Metalloproteinases-9 and Intercellular Adhesion Molecule -1 in the Embryo and Endometrium Co-Culture System

doi:10.3877/cma.j.issn.1673-5250.2012.02.011

Objective

To research the effects of interleukin(IL)-1β and IL-1Rα on the embryo and endometrium co-culture system.

Methods

A total of 30 female Kunming white mice (6 to 8 weeks) and 15 male mice (12 to 16 weeks) were selected. 8 cell stage embryos and endometrial glandular epithelial cells were collected and the endometrial cells were primarily cultured. There were 7 copies primary endometrial cells were successful cultured and were inoculated into 35 holes respectively. Control group were 5 hole cells without any treatment (n=5×7). 30 holes was divided into 6 groups according to different concentrations of IL-1β and different raio of IL-1β+ IL-1Rα. 3 groups were GⅡ with 1 ng/mL IL-1β, 10 ng/mL IL-1β and 100 ng/mL IL-1β, 3 groups were GⅡ with 1 ng/mL IL-1β+ 10 μg/mL IL-1Rα, 10 ng/mL IL-1β+ 10 μg/mL IL-1Rα and 100 ng/mL IL-1β+ 10 μg/mL IL-1Rα (n=5×7). 8 cell embryos were co-cultured with endometrial glandular epithelial cells in different media (6 embryos each hole). Supernatant were collected after 48 h co-culture, and the levels of matrix metalloproteinase-9 (MMP-9) / intercellular adhesion molecule-1 (ICAM-1) were measured by ELISA.

Results

Expressions of MMP-9 / ICAM-1 were significantly increased in 1 ng/mL IL-1β group and 10 ng/mL IL-1β group compared with those in control group respectively (P<0.05). While expression of MMP-9/ICAM-1 levels had no significant differences in 1 ng/mL IL-1β group and in 10 ng/mL IL-1β group, and also in 100 ng/mL IL-1β group and in control group (P>0.05). MMP-9/ICAM-1 levels were significant decreased in 10 ng/mL IL-1β+ 10 μg/mL IL-1Ra group compared with those control group (P<0.05); While expression of MMP-9/ICAM-1 levels had no significant differences in 1 ng/mL IL-1β+ 10 μg/mL IL-1Ra group and in control group, and also had no significant in 100 ng/mL IL-1β+ 10 μg/mL IL-1Ra group and in control group (P>0.05). They also had no significant difference in 1 ng/mL IL-1β+ 10 μg/mL IL-1Ra group compared with those in 100 ng/mL IL-1β+ 10 μg/mL IL-1Ra group (P>0.05).

Conclusions

Appropriate proportion of IL-1β and IL-1Rα can regulate embryo implantation.

Key words: interleukin-1β, interleukin-1Rα, co-culture, matrix metalloproteinase-9, intercellular adhesion molecule-1

表1 1 ng/mL，10 ng/mL与100 ng/mL IL－1β干预后共培养体系中MMP－9/ICAM－1的表达比较(±s，ng/mL)

Table 1 Comparison of expressions of MMP－9 / ICAM－1 with 1 ng/mL, 10 ng/mL and 100 ng/mL IL－1β in the co－culture system(±s，ng/mL)

组别	n	MMP－9	ICAM－1
1 ng/mL IL－1β组	35(5×7)	3.927±0.227^a	2.175±0.213^a
10 ng/mL IL－1β组	35(5×7)	4.045±0.438^ab	2.507±0.545^ab
100 ng/mL IL－1β组	35(5×7)	2.743±0.509^c	1.038±0.496^c
对照组	35(5×7)	2.418±0.517	1.313±0.382

表1 1 ng/mL，10 ng/mL与100 ng/mL IL－1β干预后共培养体系中MMP－9/ICAM－1的表达比较(±s，ng/mL)

Table 1 Comparison of expressions of MMP－9 / ICAM－1 with 1 ng/mL, 10 ng/mL and 100 ng/mL IL－1β in the co－culture system(±s，ng/mL)

组别	n	MMP－9	ICAM－1
1 ng/mL IL－1β组	35(5×7)	3.927±0.227^a	2.175±0.213^a
10 ng/mL IL－1β组	35(5×7)	4.045±0.438^ab	2.507±0.545^ab
100 ng/mL IL－1β组	35(5×7)	2.743±0.509^c	1.038±0.496^c
对照组	35(5×7)	2.418±0.517	1.313±0.382

表2 1 ng/mL，10 ng/mL与100 ng/mL IL－1β＋10 μg/mL IL－1Rα干预后共培养体系中MMP－9/ICAM－1的表达比较(±s，ng/mL)

Table 2 Comparison of expressions of MMP－9/ICAM－1(ng/mL) with 1 ng/mL, 10 ng/mL and 100 ng/mL IL－1β＋10 μg/mL IL－1Rα in the co－culture system(±s，ng/mL)

组别	n	MMP－9	ICAM－1
1 ng/mL L－1β＋10 μg/mL IL－1Rα组	35(7×5)	2.523±0.736^a	1.406±0.235^a
10 ng/mLL－1β＋10 μg/mL IL－1Rα组	35(7×5)	1.008±0.304^b	0.267±0.434^b
100 ng/mL IL－1β＋10 μg/mL IL－1Rα组	35(7×5)	2.873±0.997^ac	1.176±0.763^ac
对照组	35(7×5)	2.418±0.517	2.418±0.517

表2 1 ng/mL，10 ng/mL与100 ng/mL IL－1β＋10 μg/mL IL－1Rα干预后共培养体系中MMP－9/ICAM－1的表达比较(±s，ng/mL)

Table 2 Comparison of expressions of MMP－9/ICAM－1(ng/mL) with 1 ng/mL, 10 ng/mL and 100 ng/mL IL－1β＋10 μg/mL IL－1Rα in the co－culture system(±s，ng/mL)

组别	n	MMP－9	ICAM－1
1 ng/mL L－1β＋10 μg/mL IL－1Rα组	35(7×5)	2.523±0.736^a	1.406±0.235^a
10 ng/mLL－1β＋10 μg/mL IL－1Rα组	35(7×5)	1.008±0.304^b	0.267±0.434^b
100 ng/mL IL－1β＋10 μg/mL IL－1Rα组	35(7×5)	2.873±0.997^ac	1.176±0.763^ac
对照组	35(7×5)	2.418±0.517	2.418±0.517

1	Bonetti TC, Salomao R, Brunialti M, et al. Cytokine and hormonal profile in serum samples of patients undergoing controlled ovarian stimulation: Interleukin－1 beta predicts ongoing pregnancy[J]. Hum Reprod, 2010, 25(8):2101-2106.
2	Walker CG, Meier S, Littlejohn MD, et al. Modulation of the maternal immune system by the pre－implantation embryo[J]. BMC Genomics, 2010, 11(8):474-487.
3	Jasper MJ, Care AS, Sullivan B, et al. Macrophage－derived LIF and IL1B regulate alpha (1，2) fucosyltransferase2 (Fut2) expression in mouse uterine epithelial cells during early pregnancy [J]. Biol Reprod, 2011, 84(1):179-188.
4	Gonzalez M, Neufeld J, Reimann K, et al. Expansion of human trophoblastic spheroids is promoted by decidualized endometrial stromal cells and enhanced by heparin－binding epidermal growth factor－like growth factor and interleukin－1 β[J]. Mol Hum Reprod, 2011, 17(7):421-433.
5	Hogan BF, Lacy CE. Manipulating the mouse embryo: A laboratory manual [M]. NY：Cold Spring Harbor Laboratory Press，2003, 221-223.
6	Ratnam S, Mertineit C, Ding F, et al. Dynamics of dnmt1 methyl transferase expression and intracellular localization during oogenesis and preimplantation development[J]. Dev Biol, 2002, 245(2):304-314.
7	Liu P, Hao CF, Sheng XF, et al. Role of interleukin－1β in regulating the expression of matrix metalloproteinase－9 and intercellular adhesion molecule－1 in cultured mouse endometrial epithelial cells taken during the implantation window[J]. J Reprod Med, 2010, 12(19):514-517.
8	Raga F, Casa EM, Bonilla-Musoles F．Gonadotropin－releasing hormone (GnRH)－Ⅰ regulation of interleukin (IL)－1b and IL－1 receptor antagonist expression in cultured human endometrial stromal cells[J]. J Obstet Gynaecol Res, 2008, 34(4):464-472.
9	Simon C, Frances A, Piquette GN. Embryonic implantation in mice is blocked by interleukin－1 receptor antagonist[J]. Endocrinology, 1994, 134(2):521-528.
10	Li L, Xing FQ. Role of interleukin－1β in regulating human cultured endometrial cell MMP－9 and TIMP－3 expressions in the mid－secretory phase[J]. J Southern Med Univ, 2006, 26(8):1143-1145.
11	Borgatti M, Rizzo R, Canto MB, et al. Release of sICAM－1 in oocytes and in vitro fertilized human embryos[J]. PLoS One, 2008, 3(12):e3970.
12	Yurdakan G, Ekem TE, Bahadir B, et al. Expression of adhesion molecules in first trimester spontaneous abortions and their role in abortion pathogenesis [J]. Acta Obstet Gynecol Scand, 2008, 87(7):775-782.
13	Fontana VA, Sanchez M, Cebral E, et al.Interleukin－1 beta regulates metalloproteinase activity and leptin secretion in a cytotrophoblast model[J]. Biocell, 2010, 34(1):37-43.

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