pcDNA3-KiSS-1质粒致孕鼠妊娠高血压综合征样病变的实验研究

›› 2005, Vol. 25 ›› Issue (2): 77-82.

pcDNA3-KiSS-1质粒致孕鼠妊娠高血压综合征样病变的实验研究

1.苏州大学附属第二医院妇产科,苏州 215004；2.上海第二医科大学附属仁济医院妇产科,上海 200001

收稿日期:2004-12-21 出版日期:2005-01-25 发布日期:2013-03-19
基金资助:
国家自然科学基金(30271362)

pcDNA3-KiSS-1-Induced Pregnant Rats Exhibit Signs and Symptoms Mimicking Preeclampsia

1.Department of Obstetric and Gynecology, The Second Hospital Affiliated Suzhou University, Suzhou 215006,China；2.Department of Obstetric and Gynecology,Renji Hospital Affiliated Second Shanghai Medical University,Shanghai 200001, China

Received:2004-12-21 Online:2005-01-25 Published:2013-03-19

摘要/Abstract

摘要： 目的探讨肿瘤转移抑制基因KiSS-1表达增高抑制滋养细胞浸润及其在大鼠妊娠高血压综合征（妊高征）发病中的作用。方法妊娠Wistar大鼠随机分为A、B、C三组，每组10只，A组（实验组）于妊娠0.5、7.5、14.5d腹腔注射脂质体包埋的pcDNA3-KiSS-1质粒，B组（空质粒转染组）于同期腹腔注射脂质体包埋的pcDNA3空质粒，C组（对照组）同期腹腔注射生理盐水，观测血压、尿蛋白。于妊娠21.5d剖宫产，计数幼仔数量，并分别称取幼仔体重及胎盘重量，RTPCR检测胎盘KiSS-1 mRNA、基质金属蛋白酶9（MMP-9）mRNA表达，并取大鼠胎盘、胎盘附着处子宫壁、肾、心、肝等脏器进行病理检查。结果（1）A组妊娠大鼠于妊娠15.5 d起血压增高（P＜0.05），并于妊娠17.5 d及19.5d最为明显（P＜0.01），与B组、C组同期相比差异有统计学意义（P＜0.01）；（2）A组大鼠于妊娠14.5d出现尿蛋白升高并持续至分娩，显著高于B组及C组（P＜0.01）；（3）A组孕鼠所产仔鼠总数及体重均较B组及C组降低（P＜0.05），并且A组中有2例剖宫产时发现有流产胚胎；（4）经RT-PCR检测到A组10例大鼠中有7例其胎盘KiSS-1mRNA表达阳性，而B组及C组均为阴性，A组胎盘MMP-9mRNA表达显著低于B组及C组（P＜0.05）；（5）病理学检查结果提示A组大鼠肾小球肿胀，血管壁内皮细胞增生，肾小囊腔缩小，肾小动脉管壁增厚，管腔狭窄。三组胎盘、胎盘附着处子宫壁、心、肝等脏器未见明显差别。结论脂质体包埋的pcDNA3-KiSS-1质粒经整合至妊娠Wistar大鼠可在胎盘表达，出现血压升高、尿蛋白增高等妊高征样变化，并且胎盘MMP-9mRNA表达下降，提示KiSS-1基因表达增高导致滋养细胞浸润不足影响胎盘血管的生理性重铸，在妊高征发病中起重要作用。

关键词: KiSS-1基因, 妊娠高血压综合征, 滋养细胞, 浸润

Abstract: Objective To investigate inhibition effect of KiSS-1 gene on the trophoblast cell in order to explore the role of high expression of KiSS-1 gene in pregnancy induced hypertension syndrome. Methods Thirty pregnant Wistar rats were divided into three groups randomly with 10 rats in each group. Group A （pcDNA3-KiSS-1 treated group） was transferred recombinant eukaryotic expression vector pcDNA3-KiSS-l encapsulated with lipofectamine by intraperitoneal injection on day 0.5, day 7.5 and day 14.5 of gestation respectively. Group B （pcDNA3 treated group） was transferred pcDNA3 encapsulated with lipofectamine by intraperitoneal injection at the same day. Group C （control group） was injected 0.9% sodium chloride solution at the same day. Blood pressure and urinary protein were assessed serially. All rats were sacrificed on day 21.5 of gestation. Amount of pup, pup weight and weight of placenta were measured. Expression of KiSS-lmRNA and MMP-9niRNA in placenta were determined by reverse transcription- polymerase chain reaction （RT-PCR）.Placenta, uterus attached to placenta, kidney, heart and liver were examined histologically. Results（1） Compared with Group B and Group C, Blood pressure in Group A began rise in day 15.5 of gestation （P＜0.05）, gradually increased to peak levels until delivery. （2）Group A show increased urinary protein in day 14.5 of gestation and persisted to delivery. Daily urinary protein in Group was significantly higher than that in Group B and C （P＜0.01） (3) Pup weight in Group A was significantly lighter than that in Group B and C （P＜0.05）, and resorbed pups were observed in Group A.（4） Expression of KiSS-lmRNA was positive in 7 of 10 placenta in Group A, all in Group B and C was negative. Expression of MMP-9mRNA in placenta was significantly decreased in Group A compared with that of Group B and C （P＜0.05）.（5）Histological exami-nation show proliferation of the glomerular tufts and thickening of media of the kidney arteriolar wall in Group A compared with Group B and C. There was no significant difference in placenta, uterus attachment to placenta, heart, liver among three group. Conclusion The pcDNA3-KiSS-l vector encapsulated by lipofectamine was transfected and integrated into pregnancy Wistar rats. Expression of KiSS-lmRNA was positive in 7 of 10 pcDNA3-KiSS-1-treated rats while negative in pcDNA3-treated and control. pcDNA3-KiSS-1-treated rats exhibited the feature of preeclampsia including hypertension and proteinuria. It suggests that over expression of KiSS-1 cause deficiency of invasion of trophoblast and insufficient of remodeling of spiral arteries in placenta. It may play main role in shallow implantation of placenta in pregnancy induced hypertension syndrome.

Key words: KiSS-1 gene, Pregnancy induced hypertension, Trophoblast, Invasion

张弘1, 林其德2. pcDNA3-KiSS-1质粒致孕鼠妊娠高血压综合征样病变的实验研究[J]. , 2005, 25(2): 77-82.

ZHANG Hong-1, LIN Qi-De-2. pcDNA3-KiSS-1-Induced Pregnant Rats Exhibit Signs and Symptoms Mimicking Preeclampsia[J]. , 2005, 25(2): 77-82.

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