Department of Ophthalmology, Otorhinolaryngology and Head and Neck Surgery, Ribeirão Preto Medical School, University of São Paulo, Brazil.
Invest Ophthalmol Vis Sci. 2018 Dec 3;59(15):6036-6044. doi: 10.1167/iovs.17-22322.
The aims of this study were (1) to determine the efficacy of adenovirus vector serotype 5 (Ad) encoding human soluble VEGF receptor 1 (s-VEGFR1) gene transfer to the lacrimal gland (LG); (2) to investigate whether expression of s-VEGFR1 prevents corneal neovascularization (CNV) induced by alkali burns; and (3) to evaluate the safety of the procedure.
AdVEGFR1 vectors (25 μL, 1 × 1010 pfu/mL) were injected in the right LGs of rats and were compared with AdNull vector (25 μL, 1 × 1010 pfu/mL) or 25 μL of saline (Control) before cornea alkali burns with 1 M NaOH. After 7 days, CNV was documented at the slit lamp. Tear secretion was measured with phenol red threads. The animals were tested for s-VEGFR1 mRNA and protein in the LG by quantitative (q)PCR and immunohistochemistry staining, respectively. qPCR was used to compare the mRNA levels of IL-1β, IL-6, and TNF-α in the LG and ipsilateral trigeminal ganglion (TG).
Ad-VEGFR1 transfected 83% (10/12) of the rats. VEGFR1 was present in LG acinar cells. CNV was prevented in 9 of 12 animals in the Ad-VEGFR1 group, compared with the Ad-Null (3:10) and Control groups (1:10) (P = 0.0317). The tear secretion and cytokine mRNA levels in the LG and TG were similar in all three groups (P > 0.05).
Adenoviral vector gene transfer was safe for LG structure and function. The LG as the target tissue showed local expression of human s-VEGFR1, and CNV was prevented in most of the eyes exposed to alkali burns.
本研究旨在：（1）确定腺病毒载体 5 型（Ad）转染人可溶性血管内皮生长因子受体 1（s-VEGFR1）基因至泪腺（LG）的疗效；（2）探讨 s-VEGFR1 的表达是否能预防碱烧伤诱导的角膜新生血管化（CNV）；以及（3）评估该程序的安全性。
AdVEGFR1 载体（25 μL，1×1010 pfu/mL）注入大鼠右侧 LG，与 AdNull 载体（25 μL，1×1010 pfu/mL）或生理盐水（对照组，25 μL）进行比较，之后在角膜上用 1 M NaOH 烧伤。伤后 7 天，裂隙灯观察 CNV。用苯酚红丝测量泪液分泌。通过定量（q）PCR 和免疫组化染色分别检测 LG 中 s-VEGFR1 mRNA 和蛋白。qPCR 用于比较 LG 和同侧三叉神经节（TG）中 IL-1β、IL-6 和 TNF-α的 mRNA 水平。
Ad-VEGFR1 转染了 12 只大鼠中的 10 只（83%）。VEGFR1 存在于 LG 腺泡细胞中。与 Ad-Null（3:10）和对照组（1:10）相比，Ad-VEGFR1 组 12 只动物中有 9 只（9/12）预防了 CNV（P=0.0317）。三组大鼠的 LG 和 TG 的泪液分泌和细胞因子 mRNA 水平均无差异（P＞0.05）。
腺病毒载体基因转移对 LG 结构和功能是安全的。LG 作为靶组织，局部表达人 s-VEGFR1，大多数暴露于碱烧伤的眼睛均预防了 CNV。
