白细胞介素－1β，－1Rα对胚胎－内膜共培养体系分泌基质金属蛋白酶－9/细胞间黏附分子－1的影响

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

中华妇幼临床医学杂志(电子版) ›› 2012, Vol. 08 ›› Issue (02) : 159 -162. doi: 10.3877/cma.j.issn.1673-5250.2012.02.011

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白细胞介素－1β，－1Rα对胚胎－内膜共培养体系分泌基质金属蛋白酶－9/细胞间黏附分子－1的影响

张宁¹, 刘萍², 郝翠芳¹^,^*^,^*(

), 王昕荣¹

^1. 山东烟台　264000，烟台毓璜顶医院　生殖医学中心
^2. 枣庄市妇幼保健院

收稿日期:2012-01-03 修回日期:2012-02-28 出版日期:2012-04-01
通信作者: 郝翠芳

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

Ning ZHANG¹, Pin LIU², Cui-fang HAO¹(), Xin-rong WANG¹

^1. Reproductive Medical Center, Yantai Yuhuangding Hospital, Yantai 264000, Shandong Province, China

Received:2012-01-03 Revised:2012-02-28 Published:2012-04-01
Corresponding author: Cui-fang HAO
About author:
Corresponding author: HAO Cui-fang, Email: yyyhcf@yahoo.com.cn

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张宁, 刘萍, 郝翠芳, 王昕荣. 白细胞介素－1β，－1Rα对胚胎－内膜共培养体系分泌基质金属蛋白酶－9/细胞间黏附分子－1的影响[J]. 中华妇幼临床医学杂志(电子版), 2012, 08(02): 159-162.

Ning ZHANG, Pin LIU, Cui-fang HAO, Xin-rong WANG. 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[J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2012, 08(02): 159-162.

目的

探讨白细胞介素(IL)－1β，－1Rα在胚胎－内膜共培养体系中的作用。

方法

选取昆明种雌性小白鼠30只(6～8周龄)，雄性15只(12～16周龄)进行实验。30只雌鼠成功妊娠后，收集雌鼠8细胞期胚胎并进行子宫内膜腺上皮细胞原代培养。共培养成功7份原代子宫内膜细胞，每份细胞分别接种35孔(n＝35×7)。5孔细胞培养液中不做任何处理(对照组，n＝5×7)，其余30孔根据上皮细胞培养液中所含IL－1β浓度不同以及IL－1β ＋ IL－1Rα比例不同共分为6组，分别为1 ng/mL IL－1β组，10 ng/mL IL－1β组和100 ng/mL IL－1β组，以及1 ng/mL IL－1β＋10 μg/mL IL－1Rα组，10 ng/mL IL－1β＋10 μg/mL IL－1Rα组，100 ng/mL IL－1β＋10 μg/mL IL－1Rα组(每组样本量：n＝5×7)。将发育正常的8细胞胚胎随机置入含以上不同培养液的细胞孔内，每孔6枚胚胎，培养48 h后收集培养上清液，采用酶联免疫吸附测定(ELISA)法检测培养液中分泌蛋白基质金属蛋白酶(MMP)－9/细胞间黏附分子(ICAM)－1的表达水平。

结果

1 ng/mL IL－1β组与10 ng/mL IL－1β组，可显著促进共培养体系MMP－9/ICAM－1的表达，与对照组比较，差异有统计学意义(P<0.05)；而1 ng/mL IL－1β组与10 ng/mL IL－1β组，以及100 ng/mL IL－1β组分别与对照组比较，差异均无统计学意义(P>0.05)。10 ng/mL IL－1β＋10 μg/mL IL－1Rα组MMP－9/ICAM－1水平显著低于对照组(P<0.05)；而1 ng/mL IL－1β＋10 μg/mL IL－1Rα组，100 ng/mLIL－1β＋10 μg/mL IL－1Rα组分别与对照组比较，差异均无统计学意义(P>0.05)；1 ng/mL IL－1β＋10 μg/mL IL－1Rα组与100 ng/mL IL－1β＋10 μg/mL IL－1Rα组比较，差异亦无统计学意义(P>0.05)。

结论

IL－1β/IL－1Rα的适当比例，可以调控胚胎着床。

关键词: 白细胞介素－1β, 白细胞介素－1Rα, 共培养, 基质金属蛋白酶－9, 细胞黏附分子－1

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

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