Abstract：
Vasohibin-2 (VASH2) is confirmed to be associated with angiogenesis. To investigate the vitreous levels of VASH2 and how VASH2 induces angiogenesis in proliferative diabetic retinopathy (PDR), a total of 120 eyes were enrolled in this prospective and randomized controlled study and the vitreous level of VASH2 was quantified by Luminex liquid suspension chip. Vector systems were applied in human retinal microvascular endothelial cells (HRMECs) for VASH2 gene overexpression, along with interfering lentiviral vectors (VASH2-shRNA) for VASH2 gene silencing. Cell migration, autophagic flux, as well as the expression of α-tubulin, detyrosinated ⍺-tubulin, LC3 II/LC3 I, P62 were detected under normal, VASH2 overexpression, or interference conditions. The level of VASH2 in PDR patients was significantly higher (218.61 ± 30.14 pg/ml) than that in ERM/MH patients (80.78 ± 2.05 pg/ml) (P = 0.001). The migration ability of HRMECs was significantly increased in VASH2 overexpression group, while in the interfering group, the migration ability decreased. VASH2 increased the detyrosination of ⍺-tubulin. The high fluorescence intensity of autophagic flux showed an activation of autophagy in VASH2 overexpression group, which was also confirmed by the increase of LC3 II/LC3 I ratio and the decrease of P62. Collectively, the present study shows in PDR, vitreous level of VASH2 is higher. VASH2 promotes neovascularization by inducing autophagy, suggesting VASH2 could be a new anti-angiogenic drug target for PDR.
摘要：
Vasohibin-2(VASH2)被证实与血管生成相关。探讨VASH2在增生性糖尿病视网膜病变(PDR)中的玻璃体水平及VASH2诱导血管生成的作用,这项前瞻性随机对照研究共纳入120只眼,通过Luminex液体悬浮芯片对玻璃体VASH2水平进行了定量.载体系统应用于人视网膜微血管内皮细胞（HRMEC）中VASH2基因过表达，以及用于VASH2基因沉默的干扰慢病毒载体(VASH2-shRNA)。细胞迁移,自噬通量，以及α-微管蛋白的表达，去酪氨酸化^-微管蛋白，LC3II/LC3I,在正常情况下检测到P62，VASH2过表达,或干扰条件。PDR患者的VASH2水平（218.61±30.14pg/ml）明显高于ERM/MH患者（80.78±2.05pg/ml）（P=0.001）。VASH2过表达组HRMECs的迁移能力显著增强,在干扰组中，迁移能力下降。VASH2增加了α-微管蛋白的去酪氨酸作用。高荧光强度的自噬通量在VASH2过表达组显示自噬激活,LC3II/LC3I比值的增加和P62的降低也证实了这一点。总的来说，本研究在PDR中显示，VASH2的玻璃体水平较高。VASH2通过诱导自噬促进新生血管,提示VASH2可能是PDR新的抗血管生成药物靶点。
