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| 妊娠期接触糖皮质激素对子代大鼠心肌血清和糖皮质激素调节的蛋白激酶1表达和心脏功能的印迹效应 |
| 张治宇1,李杰2,左乔3,高颖4,徐永君4,倪鑫4,高路4* |
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(1. 第二军医大学长征医院贵宾诊疗科, 上海 200003;
2. 中央军委机关事务管理总局保健处, 北京 100034;
3. 第二军医大学长海医院神经外科, 上海 200433;
4. 第二军医大学基础部生理学教研室, 上海 200433
*通信作者) |
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| 摘要: |
| 目的 揭示妊娠期暴露于糖皮质激素(glucocorticoids,GCs)对子代大鼠心脏功能的印迹效应,探索GCs对子代心肌保护性因子的调节作用及其可能机制。方法 构建妊娠后期GCs暴露大鼠模型,通过2,3,5-氯化三苯基四氮唑(TTC)染色观察子代心肌缺血再灌注损伤后的梗死程度;实时定量PCR检测心肌保护因子血清和糖皮质激素调节的蛋白激酶1(Sgk1)、促肾上腺皮质激素释放激素(CRH)受体2(Crhr2)、CRH家族肽urocortin (Ucn)和Ucn2等基因的表达; 蛋白质印迹分析检测SGK1蛋白的表达;亚硫酸氢盐处理后测序确定Sgk1基因启动子的甲基化水平。结果 妊娠期GCs暴露大鼠子代出生时和成年后体质量减轻(P<0.01),子代雄性大鼠心肌对缺血再灌注损伤的耐受性下降(P<0.01);SGK1的 mRNA和蛋白在子代雄性大鼠心肌中的表达降低(P<0.01或P<0.05);Ucn和Ucn2在子代雌性大鼠心肌中的表达下降(P<0.05);Sgk1基因启动子区域存在多个CpG岛,且近端CpG岛的甲基化水平在妊娠期GCs暴露的子代雄性大鼠心肌中升高(P<0.01)。结论 妊娠期GCs暴露可能通过下调心肌保护因子SGK1的表达对子代雄性大鼠心肌功能产生印迹效应,而Sgk1基因启动子区域CpG岛的高甲基化改变可能是介导GCs对Sgk1的印迹效应的重要机制。 |
| 关键词: 糖皮质激素类 印迹效应 血清和糖皮质激素调节的蛋白激酶 心肌 甲基化 |
| DOI：10.16781/j.0258-879x.2016.06.0668 |
| 投稿时间：2015-09-04修订日期：2016-04-13 |
| 基金项目:国家自然科学基金面上项目(81270756). |
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| Programming effect of glucocorticoid exposure during rat pregnancy on cardiac functions and expression of serum- and glucocorticoid-regulated protein kinase 1 in offspring |
| ZHANG Zhi-yu1,LI Jie2,ZUO Qiao3,GAO Ying4,XU Yong-jun4,NI Xin4,GAO Lu4* |
(1. VIP Clinic, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China;
2. Health Office of the Administration Organization Affairs, Central Military Commission of People's Liberation Army, Beijing 100034, China;
3. Department of Neurosurgery, Changhai Hospital, Second Military Medical University, Shanghai 200433, China;
4. Department of Physiology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China
*Corresponding author) |
| Abstract: |
| Objective To investigate the programming effects of glucocorticoids (GCs) exposure during rat pregnancy on the cardiac functions of adult offspring, so as to explore the cardiac protective effect of GCs and the underlying mechanisms. Methods Advanced pregnancy GCs exposure model was established with rats. The infarction degrees of myocardium of offspring rats were evaluated by 2,3,5-triphenyltetrazolium chloride (TTC) staining after ischemia-reperfusion (I/R) injury. The mRNA levels of cardio-protective factors serum- and glucocorticoid-regulated protein kinase 1(Sgk1), corticotropin-releasing hormone receptor 2 (Crhr2), urocortin (Ucn), and Ucn2 were determined by real-time PCR. The protein level of SGK1 was detected using Western blotting analysis. Bisulfite sequencing PCR (BSP) was employed to determine the methylation level of Sgk1 promoter. Results The body mass of the offsprings of GCs-exposed pregnant rats were significantly lower than that of the normal saline-injected pregnant rats (P<0.01). The ratio between infarction area and risk area of hearts after ischemia-reperfusion in the male adult offspring from GCs-exposed group was significantly larger than that from the vehicle group(P<0.01). The mRNA and protein levels of SGK1 were significantly decreased in male adult offspring hearts exposed to GCs prenatally (P<0.01, P<0.05), whereas Ucn and Ucn2 mRNA expressions were significantly decreased in the hearts of female adult offspring exposed to GCs prenatally (P<0.05). There were multiple CpG islands in Sgk1 promoter, with the proximal CpG island in the Sgk1 promoter being significantly hypermethylated in the heart of adult male offspring exposed to GCs during late pregnancy (P<0.01). Conclusion GCs exposure during pregnancy can cause programming effects on cardiac functions of male adult offspring in rats, probably via the down-regulation of SGK1 expression in the heart, which is largely due to the hypermethylation on Sgk1 promoter. |
| Key words: glucocorticoids programming effect serum- and glucocorticoid-regulated kinase myocardium methylation |
