应用两种不同建模方式构建子痫前期SD大鼠动物模型的实验研究

Experimental Analysis of SD Rat Model of Pre-eclampsia by Using Two Different Modeling Methods

目的:探讨分析应用两种不同建模方式来构建子痫前期SD大鼠的动物模型的特异性。方法:SD孕鼠随机均分3组,空白对照组同前饲养,合体滋养细胞微绒毛(STBM)组用STBM诱导,STBM+亚硝基左旋精氨酸甲酯(L-NAME)组用STBM+L-NAME诱导。检测蛋白尿(U-TP)、血压(BP)、部分凝血活酶时间(APTT)、凝血酶原时间(PT)和D-二聚体(D-D),HE染色胎盘组织,Real time PCR和Western blot测胎盘组织可溶性血管内皮生长因子受体-1(s Flt-1)、可溶性内皮因子(SEng)、胎盘生长因子(PLGF)mRNA及蛋白表达,滋养细胞分离、培养后行功能学实验(EDU测增殖、FCM测凋亡、Transwell测侵袭)。结果:U-TP、BP、APTT、PT及D-D在孕10天3组比较差异无统计学意义(P>0.05),孕15天及20天U-TP、BP STBM+L-NAME组较STBM组显著增加(P<0.05),孕20天APTT、PT STBM+L-NAME组较STBM组显著缩短,D-D显著增加,空白对照组无明显变化。STBM+L-NAME组HE染色见胎盘蜕膜带水肿明显,炎性细胞浸润,基带和迷路带见滋养细胞增生。3组比较s Flt-1、SEng、PLGF mRNA及蛋白表达差异有统计学意义(P<0.05)。STBM+L-NAME组s Flt-1、SEng mRNA及蛋白比STBM组明显增高(P<0.05),PLGF mRNA及蛋白明显减低(P<0.05)。功能学实验中3组细胞增殖和凋亡差异均有统计学意义(P<0.05)。STBM+L-NAME组滋养细胞增殖和侵袭最少,凋亡最多。与STBM组比较差异有统计学意义(P<0.05)。结论:在构建子痫前期动物模型中,STBM合用L-NAME诱导比单独应用STBM特异性更强,为临床子痫前期诊疗提供可靠模式研究依据。

Objective：This study was aimed to compare the advantages and specifity of two different models to construct the animal model of preeclampsia in prague-Dawley（SD） rats. Methods：Female pregnancy SD rats were randomly divided into three groups（ n =30 per group） ： Control group, STBM group and L-NAME ＋ STBM group. The model of preeclampsia（PE） was established via intravenous or subcutaneous injection of STBM or L-NAME ＋ STBM. In the STBM group,STBM were injected via the vena caudalis. In the L-NAME ＋ STBM group, Nw-nitro-L-arginine-methyl ester（L-NAME） was injected subcutaneously. Control group got no treatment. Protein- uria（ U-TP）, blood pressure（ BP）, coagulation parameters （ APTT, PT, and D-D） were tested at different times. Placental sections were histopathologically examined by H-E stanning. SFIt-1 ,SEng, PLGF mRNA and protein ex- pression in placenta were tested by Real time PCR and Western blot respectively. Placental trophoblast cells were isolated and cultured,for advanced functional experiment. Results：There was no significant difference in U-TP, BP,APTT,PT and D-D between the 3 groups on pregnant 10D（P〉0.05） ,when compared with STBM group, U-TP,APTT, PT and D-D were significantly increased in STBM ＋ L-NAME group on pregnant 15 d and 20 d（ P 〈 0.05） ,and there was no significant change in control group. The expression of SFIt-1, SEng mRNA and protein were significantly different between the 3 groups （ P 〈 0, 05）. The expression of sFIt-1, SEng mRNA and protein were significantly higher in STBM ＋ L-NAME groupthan that in STBM group（ P 〈 0.05） ,with sigficantly reduced PLGF mRNA and protein in STBM ＋ L-NAME group（ P 〈 0. 05）. There was a significant difference between the 3 groups in functional experiment（ P 〈0.05）. The cell proliferation and invasion rate was lowest and the apoptosis rate was highest,with significant difference when compared to those in STBM group （ P 〈 0. 05）. Conclusions： STBM combined with L-NAME was more specific than STBM alone,to construct the model of preeclampsia in SD rats and provide a reliable basis for clinical diagnosis and treatment of preeclampsia.