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腺苷A1受体在妊娠期高血压以及子痫前期中的作用
张雪蕊1,胡建国1,战玉峰1,孟莹1,邓钦尹2,董晓静1*
0
(1. 重庆医科大学附属第二医院妇产科, 重庆 400010;
2. 重庆医科大学附属第一医院妇产科, 重庆 400016
*通信作者)
摘要:
目的 探讨腺苷A1受体(A1R)在妊娠期高血压以及子痫前期发病机制中的作用。方法 取剖宫产分娩的妊娠期高血压(n=6)、子痫前期(n=14)和正常对照(n=15)产妇胎盘组织,采用免疫荧光检测A1R在胎盘组织中的表达部位,蛋白质印迹法检测A1R在胎盘中的表达量。建立常氧、缺氧(8、24、48、72 h)以及缺氧复氧(8 h)滋养细胞模型,运用实时荧光定量PCR和蛋白质印迹法检测A1R在细胞中的表达情况,Transwell和蛋白质印迹法检测加入 A1R 拮抗剂8-环戊基-1,3-二丙基黄嘌呤(DPCPX)后对缺氧的胎盘滋养细胞侵袭性和蛋白激酶A(PKA)表达的影响。结果 A1R在胎盘表面的滋养细胞以及内皮细胞中均有表达。与正常胎盘比较,A1R在轻度和重度子痫前期胎盘组织中表达升高(P<0.01)。A1R在体外胎盘滋养细胞缺氧模型中的表达随着缺氧时间的延长而逐渐升高(P<0.05)。胎盘滋养细胞缺氧模型中滋养细胞的侵袭能力较正常组下降(P<0.01),PKA表达下降(P<0.05);加入拮抗剂DPCPX可以增强滋养细胞的侵袭性(P<0.05),同时上调PKA的表达水平(P<0.05)。结论 缺氧程度的增加可以使A1R表达升高,A1R在胎盘滋养细胞中的作用机制可能与调节PKA相关。
关键词:  腺苷A1受体  子痫前期  胎盘  滋养细胞  缺氧  蛋白激酶A
DOI:10.16781/j.0258-879x.2017.03.0323
投稿时间:2016-06-28修订日期:2016-09-05
基金项目:
Role of adenosine A1 receptor in gestational hypertension and preeclampsia
ZHANG Xue-rui1,HU Jian-guo1,ZHAN Yu-feng1,MENG Ying1,DENG Qin-yin2,DONG Xiao-jing1*
(1. Department of Obstetrics and Gynecology, the Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China;
2. Department of Obstetrics and Gynecology, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
*Corresponding author)
Abstract:
Objective To explore the role of adenosine A1 receptor (A1R) in pathogenesis of gestational hypertension and preeclampsia. Methods Placenta tissues derived from puerperae with gestational hypertension (n=6), preeclampsia (n=14) and from normal control placentae (n=15) were obtained. The expression location of A1R protein in placenta tissues was detected by immunofluorescence technique, and the expression quantity of A1R protein was performed by Western blotting analysis. We created the normal, hypoxia (8, 24, 48 and 72 h) and hypoxia-reoxygenation trophoblastic cell models, and detected the A1R mRNA and protein expressions in trophoblastic cell models by real-time quantification PCR and Western blotting analysis. Transwell assay was used to determine the invasion ability of trophoblastic cells treated with the A1R antagonist, 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), and Western blotting analysis was used to detect the expression of protein kinase A (PKA) protein. Results We observed that A1R protein was expressed in both trophoblastic cells and vascular endothelial cells. A1R expression was significantly higher in mild and severe preeclampsia placenta tissues than that in normal placenta tissues (P<0.01). A1R expression in the hypoxia model of trophoblast cells significantly was increased with the prolongation of hypoxia (P<0.05). The invasion ability and PKA expression in the hypoxia model of trophoblast cells were significantly lower than those in normal placenta tissues (P<0.01, P<0.05). DPCPX significantly increased the invasion ability of trophoblastic cells and the PKA expression (P<0.05). Conclusion The prolongation of hypoxia can increase the expression of A1R in the trophoblast cells, which may be related to regulated PKA expression.
Key words:  adenosine A1 receptor  pre-eclampsia  placenta  trophoblast cells  hypoxia  protein kinase A