Zhang Bo, Wang Dan, Ji Tie-Feng, Shi Lei, Yu Jin-Lu
Department of Neurosurgery, The First Hospital of Jilin University, Changchun, P.R. China.
Department of Ophthalmology, The First Hospital of Jilin University, Changchun, P.R. China.
Oncotarget. 2017 Mar 7;8(10):17347-17359. doi: 10.18632/oncotarget.14468.
This study aimed to explore the effects of lncRNA ANRIL on vascular endothelial growth factor (VEGF) and angiogenesis in diabetes mellitus (DM) combined with cerebral infarction (CI) through NF-κB signaling pathway.
Adult male Wistar rats were randomly divided into control group and DM + CI group, and the DM + CI group were subdivided into Vector, shANRIL, PDTC, pcDNA-ANRIL, and pcDNA-ANRIL + PDTC groups. VEGF and FMS-like tyrosine kinase (FLT-1) expressions were measured by immunohistochemistry and endothelium dependent microvessel density (MVD) was detected by differentiation 31 (CD31) and para-amiuosalicylic acid (PAS) double staining. The qRT-PCR was applied to measure mRNA expressions of VEGF, FLT-1, Kinase insert domain protein receptor (FLK-1) and NF-κB, and Western blotting was conducted to detected expressions of VEGF, NF-κB and p-IκB/IκB.
Compared with the control group, protein expressions of VEGF, NF-κB, p-IκB/IκB, expression of ANRIL, and mRNA expressions of VEGF, FLT-1 and NF-κB were increased in the DM + CI group. Compared with the Vector group, protein expressions of VEGF, NF-κB, p-IκB/IκB, expression of ANRIL, mRNA expressions of VEGF, FLT-1 and NF-κB, and endothelium dependent MVD were increased in the pcDNA-ANRIL group, while decreased in the shANRIL group and PDTC group. Compared with the pcDNA-ANRIL group, protein expressions of VEGF, NF-κB, p-IκB/IκB, expression of ANRIL, mRNA expressions of VEGF, FLT-1 and NF-κB, and endothelium dependent MVD were decreased in the pcDNA-ANRIL + PDTC group.
Overexpressed lncRNA ANRIL upregulates VEGF and promotes angiogenesis by activating NF-κB signaling pathway in DM + CI rats. .
本研究旨在通过核因子κB（NF-κB）信号通路探讨长链非编码RNA（lncRNA）ANRIL对糖尿病（DM）合并脑梗死（CI）中血管内皮生长因子（VEGF）及血管生成的影响。
成年雄性Wistar大鼠随机分为对照组和DM + CI组，DM + CI组再细分为载体组、shANRIL组、吡咯烷二硫代氨基甲酸盐（PDTC）组、pcDNA-ANRIL组和pcDNA-ANRIL + PDTC组。采用免疫组织化学法检测VEGF和FMS样酪氨酸激酶（FLT-1）表达，用分化簇31（CD31）和对氨基水杨酸（PAS）双重染色检测内皮依赖性微血管密度（MVD）。应用实时定量聚合酶链反应（qRT-PCR）检测VEGF、FLT-1、激酶插入结构域蛋白受体（FLK-1）和NF-κB的mRNA表达，采用蛋白质印迹法检测VEGF、NF-κB和磷酸化IκB/ IκB的表达。
与对照组相比，DM + CI组中VEGF、NF-κB、磷酸化IκB/ IκB的蛋白表达、ANRIL表达以及VEGF、FLT-1和NF-κB的mRNA表达均增加。与载体组相比，pcDNA-ANRIL组中VEGF、NF-κB、磷酸化IκB/ IκB的蛋白表达、ANRIL表达、VEGF、FLT-1和NF-κB的mRNA表达以及内皮依赖性MVD均增加，而shANRIL组和PDTC组则降低。与pcDNA-ANRIL组相比，pcDNA-ANRIL + PDTC组中VEGF、NF-κB、磷酸化IκB/ IκB的蛋白表达、ANRIL表达、VEGF、FLT-1和NF-κB的mRNA表达以及内皮依赖性MVD均降低。
在DM + CI大鼠中，lncRNA ANRIL过表达通过激活NF-κB信号通路上调VEGF并促进血管生成。
