VIP Induces Changes in the F-/G-Actin Ratio of Schlemm's Canal Endothelium via LRRK2 Transcriptional Regulation
VIP Induces Changes in the F-/G-Actin Ratio of Schlemm's Canal Endothelium via LRRK2 Transcriptional Regulation
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VIP 通过 LRRK2 转录调节诱导施累姆氏管内皮 F-/G-肌动蛋白比率的变化
DOI：
10.1167/iovs.61.6.45
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发表时间：
2020
影响因子：
4.4
通讯作者：
Chen Liwen
中科院分区：
文献类型：
--
作者：
Yan Xiaoqin;Li Mu;Luo Zhaoxia;Zhao Yin;Zhang Hong;Chen Liwen
目的研究表明，血管活性肠肽(VIP)对大鼠青光眼模型具有调节Schlemm管(SC)内皮细胞骨架和扩张SC管腔的作用。本研究旨在探讨VIP对细胞骨架调控的分子机制。方法在大鼠体内实验中，应用免疫荧光法检测VIP作用后SC周围亮氨酸重复蛋白2(LRRK2)的表达及F-肌动蛋白/G-肌动蛋白(F-/G-肌动蛋白)的比值。在人脐静脉内皮细胞的体外实验中，应用定量聚合酶链式反应(QPCR)和免疫印迹法检测VIP(和Sp1/LRRK2抑制剂)作用后Sp1和LRRK2的表达。应用免疫荧光法和免疫印迹法检测VIP(和LRRK2抑制剂)作用后F-/G-肌动蛋白比值的变化。结果VIP在体内和体外均可诱导LRRK2的表达增加，并在体外引起Sp1的核转位。Sp1抑制剂的应用可在体外阻断VIP诱导的LRRK2表达增加。此外，VIP改变了F-/G-肌动蛋白的比例，这种作用在体内和体外均被LRRK2抑制剂所阻断。结论VIP可上调LRRK2的表达，其调节机制可能与Sp1的核转位有关。VIP进一步改变了F-/G-肌动蛋白的比例，调节了F-肌动蛋白结构的稳定和失稳之间的平衡。本研究阐明了VIP通过Sp1-LRRK2途径调节SC内皮细胞肌动蛋白细胞骨架的新机制，为青光眼的治疗提供了一种新的策略。
Purpose A previous study reported that vasoactive intestinal peptide (VIP) can regulate the cytoskeleton of Schlemm's canal (SC) endothelium and expand the SC lumen in a rat glaucoma model. In this study, we aimed to investigate the molecular mechanism of VIP on cytoskeleton regulation. Methods During in vivo experiments in rats, leucine-rich repeat kinase 2 (LRRK2) expression and the ratio of F-actin to G-actin (F-/G-actin) surrounding SC were examined by immunofluorescence after the application of VIP. For in vitro experiments in human umbilical vein endothelial cells, both quantitative PCR (qPCR) and western blotting were performed to evaluate Sp1 and LRRK2 expression after the application of VIP (and Sp1/LRRK2 inhibitor). In addition, the F-/G-actin ratio was examined by both immunofluorescence and western blotting after the application of VIP (and LRRK2 inhibitor). Results VIP induced increases in the expression of LRRK2 both in vivo and in vitro and the nuclear translocation of Sp1 in vitro. The application of Sp1 inhibitor abolished the increase in LRRK2 expression induced by VIP in vitro. In addition, VIP changed the F-/G-actin ratio, and this effect was abolished by the LRRK2 inhibitor both in vivo and in vitro. Conclusions VIP increased the expression of LRRK2, and this regulation was due to the nuclear translocation of Sp1. VIP further changed the F-/G-actin ratio and regulated the balance between the stabilization and destabilization of the F-actin architecture. This study elucidates a novel mechanism by which VIP regulates the actin cytoskeleton of SC endothelium via the Sp1–LRRK2 pathway, suggesting a potential novel treatment strategy for glaucoma.