MicroRNA-376b-3p Suppresses Choroidal Neovascularization by Regulating Glutaminolysis in Endothelial Cells.
MicroRNA-376b-3p Suppresses Choroidal Neovascularization by Regulating Glutaminolysis in Endothelial Cells.
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MicroRNA-376b-3p 通过调节内皮细胞中的谷氨酰胺分解来抑制脉络膜新生血管形成
DOI：
10.1167/iovs.64.1.22
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发表时间：
2023-01-03
影响因子：
4.4
通讯作者：
中科院分区：
文献类型：
--
作者：
目的脉络膜新生血管（CNV）是多种眼部疾病的共同病理改变，严重损害中心视力。积累的证据表明，微小RNA（miRNA）与内皮代谢的调节密切相关，在血管生成中发挥着至关重要的作用。在此，我们研究了miR-376 b-3 p在CNV进展中调节内皮谷氨酰胺代谢的分子机制。方法将miR-376 b-3 p模拟物和对照转染人视网膜微血管内皮细胞（HRMEC），采用实时荧光定量PCR和Western blotting方法检测HRMEC中谷氨酰胺酶1（GLS 1）的表达。用相关试剂盒检测转染细胞的生物学功能和谷氨酰胺代谢。荧光素酶报告基因测定用于验证CCAAT/增强子结合蛋白β（CEBPB）是miR-376 b-3 p的靶标。进行染色质免疫沉淀和RNA免疫沉淀测定以验证CEBPB对GLS 1的启动子区域的结合。荧光素眼底血管造影和免疫荧光检测miR-376 b-3 p agomir对激光诱导大鼠CNV的影响。结果CNV大鼠视网膜色素上皮-脉络膜复合体中miR-376 b-3 p表达降低，GLS 1表达升高。用miR-376 b-3 p模拟物转染的HRMEC显示出CEBPB的抑制，导致GLS 1转录失活和精氨酸分解。此外，miR-376 b-3 p模拟物抑制HRMEC的增殖、迁移和管形成，但促进凋亡，而这些作用被α-酮戊二酸补充或用CEBPB过表达质粒转染抵消。最后，玻璃体内施用miR-376 b-3 p阿戈米尔抑制CNV形成。结论miR-376 b-3 p是内皮细胞谷氨酰胺代谢的抑制因子，有望成为CNV相关疾病的治疗靶点。
Purpose Choroidal neovascularization (CNV) is a common pathological change of various ocular diseases that causes serious damage to central vision. Accumulated evidence shows that microRNAs (miRNAs) are closely related with the regulation of endothelial metabolism, which plays crucial roles in angiogenesis. Here, we investigate the molecular mechanism underlying the regulation of endothelial glutamine metabolism by miR-376b-3p in the progression of CNV. Methods Human retinal microvascular endothelial cells (HRMECs) were transfected with control or miR-376b-3p mimics, and the expression of glutaminase 1 (GLS1), a rate-limiting enzyme in glutaminolysis, was detected by real-time PCR or Western blotting. The biological function and glutamine metabolism of transfected HRMECs were measured by related kits. Luciferase reporter assays were used to validate the CCAAT/enhancer-binding protein beta (CEBPB) was a target of miR-376b-3p. Chromatin immunoprecipitation and RNA immunoprecipitation assays were performed to verify the binding of CEBPB on the promoter region of GLS1. Fundus fluorescein angiography and immunofluorescence detected the effect of miR-376b-3p agomir on rat laser-induced CNV. Results The expression of miR-376b-3p was decreased, whereas GLS1 expression was increased in the retinal pigment epithelial–choroidal complexes of rats with CNV. HRMECs transfected with miR-376b-3p mimic showed inhibition of CEBPB, resulting in the inactivation of GLS1 transcription and glutaminolysis. Moreover, the miR-376b-3p mimic inhibited proliferation, migration and tube formation but promoted apoptosis in HRMECs, whereas these effects counteracted by α-ketoglutarate supplementation or transfection with CEBPB overexpression plasmid. Finally, the intravitreal administration of the miR-376b-3p agomir restrained CNV formation. Conclusions Collectively, miR-376b-3p is a suppressor of glutamine metabolism in endothelial cells that could be expected to become a therapeutic target for the treatment of CNV-related diseases.
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影响因子：
14.9
作者：
Dreos R;Ambrosini G;Cavin Périer R;Bucher P
通讯作者：
Bucher P
影响因子：
56.9
作者：
Lagos-Quintana, M;Rauhut, R;Tuschl, T
通讯作者：
Tuschl, T
影响因子：
64.5
作者：
De Bock, Katrien;Georgiadou, Maria;Carmeliet, Peter
通讯作者：
Carmeliet, Peter
YAP 通过改变 PFKFB3 驱动的内皮糖酵解促进眼部新生血管形成
影响因子：
9.8
作者：
Feng, Yifan;Zou, Rong;Yuan, Fei
通讯作者：
Yuan, Fei
DOI：
10.26355/eurrev_202005_21170
发表时间：
2020-05-01
影响因子：
3.3
作者：
Chen, L-M;Niu, Y-D;Lin, H.
通讯作者：
Lin, H.