摘要
目的探讨高血糖对胎鼠肺组织中磷酸化雷帕霉素靶蛋白(phosphorylation mammalian target of ra-pamycin,p-mTOR)、缺氧诱导因子.1仅(hypoxia inducible factor-1α,HIF-1α)及血管内皮生长因子A(vascular endothelial growth factor-A,VEGF-A)蛋白表达的影响及意义。方法妊娠大鼠分成实验组(糖尿病组,GDM组)、干预组(糖尿病模型+雷帕霉素)及对照组(柠檬酸缓冲液)。光镜下观察各组孕21d胎鼠肺组织的大体形态结构;利用免疫组化技术检测各组胎鼠肺组织中p-mTOR、HIF-1α及VEGF-A的定位表达;Western印迹技术检测胎鼠肺组织中p-mTOR、HIF-1α、VEGF-A蛋白水平;RT.PCR技术检测胎鼠肺组织中HIF-10l、VEGF-AmRNAs水平。结果免疫组化及Western印迹结果显示,与对照组相比。糖尿病组p-mTOR、HIF-1α、VEGF-A在胎鼠肺组织中的表达水平明显增加,干预组中各蛋白表达水平与GDM组相比显著降低。RT-PCR结果显示,与对照组相比,糖尿病组中p-mTOR、HIF-1α、VEGF-AmRNAs相对平均量显著增加,干预组中各基因mRNAs水平与GDM组相比显著降低。结论妊娠期糖尿病高血糖可导致胎鼠肺组织中异常激活的p-roTOR及其调控因子HIF-1及VEGFA表达增高,可能与胎鼠肺血管的发育异常相关。
Objective investigate the effect of hyperglycemia on the expression of p-mTOR, HIF-1 and VEGF-A vascular endothelial growth factor (VEGF) in fetal rat lung. Methods Pregnant rats were divided into experimental group (diabetic group ), intervention group (diabetic model + rapamycin) and control group (citric acid buffer) . The gross morphological structure of fetal lung tissue was observed under light microscope for 21 days ; The expression of p-mTOR, HIF-1 txand VEGF-A in fetal lung tissue was detected by immunohistochemistry; The expression level of p-mTOR, HIF-1α and VEGF-A protein in fetal lung tissue was detected by Western Blot technique, and the expression level of HIF-1 ctand VEGF-A in fetal lung tissue was detected by RT-PCR tech- nique. Results Compared with the control group, the alveolar cavity decreased and the number in- creased, the alveolar septum thickened and the fetal lung structure poorly developed in the DM group and the intervention group. Immunohistochemical and Western Blot results showed that, com- pared with the control group, the expression of P-mTOR, HIF-1 aand VEGF-A in the fetal lung tis-sue was significantly increased in the diabetic group, The protein expression level of the intervention group was between the control group and the diabetes group. RT-PCR results showed that compared with the control group, the relative average of HIF-1α Land VEGF-A mRNAs in the GDM group decreased significantly, while the level of HIF-1 otand VEGF-A mRNAs in the intervention group was significantly lower than that in the GDM group. Conclusion Hyperglycemia in gestational diabetes mellitus can induce abnormal activation of p-mTOR and its regulators, HIF-1α and VEGF-A, infetal lung tissues. It may be related to abnormal development of fetal pulmonary vessels.
作者
邓飞涛
宋婷
张青苗
欧阳为相
柴新群
DENG Feitao;SONG Ting;ZHANG Qingmiao;OUYANG Weixiang;CHAI Xinqun(Department of Obstetrics and Gynecology,Union Hospital,Huazhong University of Science and Technology,Wuhan,430022,China;Department of Hepatobiliary Surgery,Union Hospital,Huazhong University of Science and Technology,Wuhan,430022,China)
出处
《医学分子生物学杂志》
CAS
2018年第5期309-314,共6页
Journal of Medical Molecular Biology
基金
国家自然科学基金(No.81070059),湖北省自然科学基金(No.2017CFB749)