不同抗氧化剂对胎膜组织MMP-9表达的影响  

Effects of different antioxidants on MMP-9 protein expression in fetal membrane tissue

作　　者：李玲[1] 崔金晖[1] 陈新娟[1] 范建辉[1] 

机构地区：[1]中山大学附属第三医院产科,广东广州510630

出　　处：《中国病理生理杂志》2014年第5期909-913,共5页Chinese Journal of Pathophysiology

基　　金：广东省科技计划(No.2011B061300030)

摘　　要：目的:检测不同抗氧化剂α-硫辛酸(α-LA)和维生素C(Vit C)+维生素E(Vit E)对体外培养的胎膜早破(PROM)胎膜组织基质金属蛋白酶9(MMP-9)表达的影响。方法:选取发生胎膜早破的孕妇8例,未临产,剖宫产终止妊娠,其中未足月胎膜早破组(PPROM)和足月胎膜早破组(TPROM)各4例。每例胎膜组织分成3组,分别加入磷酸盐缓冲液(PBS,即对照组)、0.5 mmol/Lα-LA和28.8 g/L Vit C+1.8 g/L Vit E后进行体外培养24h,采用免疫组化染色方法和Western blotting技术检测胎膜组织中MMP-9蛋白表达。结果:(1)免疫组化结果显示:TPROM组织中MMP-9定位于绒毛膜细胞和中性粒细胞的胞浆,且3组间MMP-9平均积分吸光度值无显著差异(P>0.05),而PPROM组织中MMP-9除在绒毛膜细胞和中性粒细胞的胞浆中表达外,在羊膜上皮细胞的胞浆中也强阳性表达,α-LA组及Vit C+Vit E组MMP-9积分吸光度值(0.0212±0.0057和0.0107±0.0009)低于对照组(0.0618±0.0098),Vit C+Vit E组较α-LA组更低(P<0.01)。(2)Western blotting结果显示,TPROM组织中,3组中pro-MMP-9表达水平均无显著差异(P>0.05),但α-LA组及Vit C+Vit E组active MMP-9蛋白相对表达水平(0.64±0.07和0.28±0.13)低于对照组(0.99±0.22),Vit C+Vit E组较α-LA组更低(P<0.05)。在PPROM组织中,α-LA组和Vit C+Vit E组pro-MMP-9/active MMP-9蛋白相对表达水平(0.69±0.08/0.35±0.07和0.38±0.03/0.16±0.05)低于对照组(1.00±0.20/1.00±0.10),Vit C+Vit E组较α-LA组更低(P<0.05)。结论:抗氧化剂可使胎膜早破胎膜组织的MMP-9表达水平下调,且28.8 g/L Vit C+1.8 g/L Vit E作用后的MMP-9表达水平较0.5 mmol/Lα-LA下调更明显,这为胎膜早破胎膜组织的防治提供理论依据。AIM： To find out the influences of different antioxidants on the protein expression of matrix metalloproteinase 9 （ MMP-9 ） in the membrane tissue of premature rupture cuhured in vitro. METHODS ： Eight pregnant women of premature rupture of membranes （PROM） who had delivered by cesarean section before labor were enrolled in the study and divided into 2 groups： term premature rupture of membranes （TPROM） and preterm premature rupture of membranes （PPROM）. The fetal membrane tissues from each case was collected and divided into 3 sub-groups, which were cultured for 24 h with phosphate buffered saline （ PBS, control group） , 0. 5 mmol/L alpha-lipoic acid （ α-LA ） , or 28.8 g/L vitamin C ＋ 1. 8 g/L vitamin E in vitro. The protein expression of MMP-9 was determined by immunohistochemistry and Western blotting. RESULTS： MMP-9 was located in the chorionic cells and neutrophils in TPROM tissues and there was no significant difference among samples of different treatments. In PPROM tissues, MMP-9 was found not only in the chorionic cells and neutrophils, but also in the amniotic epithelial cells. The integral absorbance values in antioxidanttreated samples were lower than that in PBS-treated samples, and the significant difference between α-LA-treated samples and vitamin C ＋ vitamin E-treated samples was observed. The expression of pro-MMP-9 in TPROM tissues showed no signif- icant difference after treated with different antioxidants, but the active MMP-9 expression was decreased after treated with antioxidants, and the influence of vitamin C ＋ vitamin E was stronger than α-LA. The expression of pro-MMP-9/active MMP-9 in PPROM tissues was decreased after treated with antioxidants, and the influence of vitamin C ＋ vitamin E was stronger than α-LA. CONCLUSION： Antioxidants result in the down-regulation of MMP-9 protein expression, and the influence of vitamin C ＋ vitamin E is stronger than α-LA, indicating that these antioxidants can be used clinically to prevent premature ru

关 键 词：胎膜早破 Α-硫辛酸 维生素C 维生素E 基质金属蛋白酶9 

分 类 号：R363[医药卫生—病理学]