Huang Jingyi, Zhang Yiran, Lin Tao, Yin Hui, Pan Yingzhe, Zhu Meijuan, Zhang Min
Department of Ophthalmology, Xiangyang No. 1 People's Hospital, Hubei University of Medicine, Xiangyang, 441000.
Xiangyang Hospital of Integrated Traditional Chinese and Western Medicine, Xiangyang, 441004.
Heliyon. 2023 Mar 31;9(4):e14869. doi: 10.1016/j.heliyon.2023.e14869. eCollection 2023 Apr.
To prepare an ophthalmic solution with a cell-permeable TAT peptide (TAT-N24) as the main cell-permeable peptide inhibitor of p55PIK signaling and observe its therapeutic effect on suture-induced corneal neovascularization (CNV) in rats. Sprague-Dawley rats were used to establish a corneal suture (CS) model of CNV. The vehicle and 0.9% TAT-N24 ophthalmic solution was topically administered. CNV induction was assessed on the basis of the clinical performance of each group. Hematoxylin-eosin staining was used to observe pathological changes, and immunohistochemical staining and confocal immunofluorescence were used to determine the localization of factors associated with corneal tissue. The mRNA expression levels of hypoxia-inducible factor (HIF-1α), vascular endothelial growth factor (VEGF-A), nuclear transcription factor κB (NF-κB p65), tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), and interleukin (IL)-6 were determined using real-time quantitative polymerase chain reaction. Western blotting was performed to detect the protein expression levels of HIF-1α and NF-κB p65. TAT-N24 slowed CNV production and reduced the expression of HIF-1α and inflammatory factors in CS models. The mRNA levels of HIF-1α, VEGF-A, NF-kB, TNF-α, IL-1β, and IL-6 significantly decreased. Moreover, the protein levels of HIF-1α and NF-κB p65 were significantly decreased. TAT-N24 can treat CNV and ocular inflammation by inhibiting the HIF-1α/NF-κB signaling pathway in CS. In the early treatment of corneal foreign body trauma, topical application of TAT-N24 can not only reduce the inflammatory response but also inhibit corneal neovascularization.
制备以细胞穿透性TAT肽(TAT-N24)作为p55PIK信号传导的主要细胞穿透性肽抑制剂的眼用溶液,并观察其对大鼠缝线诱导的角膜新生血管化(CNV)的治疗效果。采用Sprague-Dawley大鼠建立CNV的角膜缝线(CS)模型。局部给予赋形剂和0.9% TAT-N24眼用溶液。根据每组的临床表现评估CNV诱导情况。采用苏木精-伊红染色观察病理变化,采用免疫组织化学染色和共聚焦免疫荧光法确定与角膜组织相关因子的定位。使用实时定量聚合酶链反应测定缺氧诱导因子(HIF-1α)、血管内皮生长因子(VEGF-A)、核转录因子κB(NF-κB p65)、肿瘤坏死因子(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素(IL)-6的mRNA表达水平。进行蛋白质免疫印迹法检测HIF-1α和NF-κB p65的蛋白表达水平。TAT-N24减缓了CS模型中CNV的产生,并降低了HIF-1α和炎症因子的表达。HIF-1α、VEGF-A、NF-κB、TNF-α、IL-1β和IL-6的mRNA水平显著降低。此外,HIF-1α和NF-κB p65的蛋白水平也显著降低。TAT-N24可通过抑制CS中的HIF-1α/NF-κB信号通路治疗CNV和眼部炎症。在角膜异物创伤的早期治疗中,局部应用TAT-N24不仅可以减轻炎症反应,还可以抑制角膜新生血管化。