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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
张晓丽;赵晓勇
0
(南方医科大学附属花都医院)
摘要:
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。
关键词:  建模  构建  子痫前期  动物模型  分析
DOI:
基金项目:广东省科技厅科技项目(编号:2015A030302004,2015A030302003);广东省中医药局科技项目(编号:20161191,20151061);广东省自然科学基金(编号:2016A030313419)
Experimental Analysis of SD Rat Model of Pre-eclampsia by Using Two Different Modeling Methods
ZHANG Xiaoli;ZHAO Xiaoyong
(Huadu Hospital of Southern Medical University)
Abstract:
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-Larginine-methyl ester( L-NAME) was injected subcutaneously. Control group got no treatment. Proteinuria( UTP),blood pressure( BP),coagulation parameters( APTT,PT,and D-D) were tested at different times. Placental sections were histopathologically examined by H-E stanning. SFlt-1,SEng,PLGF mRNA and protein expression 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 SFlt-1,SEng mRNA and protein were significantly different between the 3 groups( P<0. 05). The expression of s Flt-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
Key words:  Modeling  Construction  Preeclampsia  Animalmodel  Analysis

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