中华妇幼临床医学杂志（电子版）
中華婦幼臨床醫學雜誌（電子版）
중화부유림상의학잡지（전자판）
CHINESE JOURNAL OF OBSTETRICS & GYNECOLOGY AND PEDIATRICS(ELECTRONIC VERSION)
2012年
2期
136-139
,共4页
张宁%刘萍%郝翠芳%王昕荣
張寧%劉萍%郝翠芳%王昕榮
장저%류평%학취방%왕흔영
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
白細胞介素-1β%白細胞介素-1Rα%共培養%基質金屬蛋白酶-9%細胞黏附分子-1
백세포개소-1β%백세포개소-1Rα%공배양%기질금속단백매-9%세포점부분자-1
interleukin-1β%interleukin-1Rα%co-culture%matrix metalloproteinase-9%intercellular adhesion molecule-1
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 IL1β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细胞胚胎随机置人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/mLIL-1Rα组与100 ng/mL IL-1β+10 μg/mL 1L-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.
目的 探討白細胞介素(IL)-1β,-1Rα在胚胎-內膜共培養體繫中的作用.方法 選取昆明種雌性小白鼠30隻(6～8週齡),雄性15隻(12～16週齡)進行實驗.30隻雌鼠成功妊娠後,收集雌鼠8細胞期胚胎併進行子宮內膜腺上皮細胞原代培養.共培養成功7份原代子宮內膜細胞,每份細胞分彆接種35孔(n=35×7).5孔細胞培養液中不做任何處理(對照組,n=5×7),其餘30孔根據上皮細胞培養液中所含IL-1β濃度不同以及IL-1β+IL-1Rα比例不同共分為6組,分彆為1 ng/mL IL1β組,10 ng/mL IL-1β組和100 ng/mL IL 1β組,以及1 ng/mL IL-1β+10 μg/mL IL-1Rα組,10 ng/mL IL 1β+10 μg/mL IL-1Rα組,100 ng/mL IL-1β+10 μg/mL IL-1Rα組(每組樣本量:n=5×7).將髮育正常的8細胞胚胎隨機置人含以上不同培養液的細胞孔內,每孔6枚胚胎,培養48 h後收集培養上清液,採用酶聯免疫吸附測定(ELISA)法檢測培養液中分泌蛋白基質金屬蛋白酶(MMP)-9/細胞間黏附分子(ICAM)-1的錶達水平.結果 1 ng/mL IL-1β組與10 ng/mL IL-1β組,可顯著促進共培養體繫MMP-9/ICAM-1的錶達,與對照組比較,差異有統計學意義(P＜0.05);而1 ng/mL IL-1β組與10 ng/mL IL-1β組,以及100 ng/mL IL-1β組分彆與對照組比較,差異均無統計學意義(P＞0.05).10 ng/mL IL-1β+ 10 μg/mL IL 1Rα組MMP-9/ICAM-1水平顯著低于對照組(P＜0.05);而1 ng/mL IL-1β+ 10 μg/mL IL-1Rα組,100 ng/mLIL-1β+10 μg/mL IL 1Rα組分彆與對照組比較,差異均無統計學意義(P＞0.05);1 ng/mL IL-1β+ 10 μg/mLIL-1Rα組與100 ng/mL IL-1β+10 μg/mL 1L-1Rα組比較,差異亦無統計學意義(P＞0.05).結論 IL-1β/IL-1Rα的適噹比例,可以調控胚胎著床.
목적 탐토백세포개소(IL)-1β,-1Rα재배태-내막공배양체계중적작용.방법 선취곤명충자성소백서30지(6～8주령),웅성15지(12～16주령)진행실험.30지자서성공임신후,수집자서8세포기배태병진행자궁내막선상피세포원대배양.공배양성공7빈원대자궁내막세포,매빈세포분별접충35공(n=35×7).5공세포배양액중불주임하처리(대조조,n=5×7),기여30공근거상피세포배양액중소함IL-1β농도불동이급IL-1β+IL-1Rα비례불동공분위6조,분별위1 ng/mL IL1β조,10 ng/mL IL-1β조화100 ng/mL IL 1β조,이급1 ng/mL IL-1β+10 μg/mL IL-1Rα조,10 ng/mL IL 1β+10 μg/mL IL-1Rα조,100 ng/mL IL-1β+10 μg/mL IL-1Rα조(매조양본량:n=5×7).장발육정상적8세포배태수궤치인함이상불동배양액적세포공내,매공6매배태,배양48 h후수집배양상청액,채용매련면역흡부측정(ELISA)법검측배양액중분비단백기질금속단백매(MMP)-9/세포간점부분자(ICAM)-1적표체수평.결과 1 ng/mL IL-1β조여10 ng/mL IL-1β조,가현저촉진공배양체계MMP-9/ICAM-1적표체,여대조조비교,차이유통계학의의(P＜0.05);이1 ng/mL IL-1β조여10 ng/mL IL-1β조,이급100 ng/mL IL-1β조분별여대조조비교,차이균무통계학의의(P＞0.05).10 ng/mL IL-1β+ 10 μg/mL IL 1Rα조MMP-9/ICAM-1수평현저저우대조조(P＜0.05);이1 ng/mL IL-1β+ 10 μg/mL IL-1Rα조,100 ng/mLIL-1β+10 μg/mL IL 1Rα조분별여대조조비교,차이균무통계학의의(P＞0.05);1 ng/mL IL-1β+ 10 μg/mLIL-1Rα조여100 ng/mL IL-1β+10 μg/mL 1L-1Rα조비교,차이역무통계학의의(P＞0.05).결론 IL-1β/IL-1Rα적괄당비례,가이조공배태착상.
Objective To research the effects of interleukin(IL) 1β3 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-1β3+IL-1Ra.3 groups were GⅡ with 1 ng/mL IL-1β,10 ng/mL IL1β and 100 ng/mL IL-1β,3 groups were G Ⅱ with 1 ng/mL IL-1β+ 10 μg/mL IL 1Rα,10 ng/mL IL-1β+ 10μg/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/ICAM1 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.
