Subconjunctival Bevacizumab Injection Impairs Corneal Innervations and Epithelial Wound Healing in Mice
Subconjunctival Bevacizumab Injection Impairs Corneal Innervations and Epithelial Wound Healing in Mice
复制标题
结膜下注射贝伐珠单抗损害小鼠角膜神经支配和上皮伤口愈合
DOI：
10.1167/iovs.16-20926
复制
发表时间：
2017-03-01
影响因子：
4.4
通讯作者：
Shi, Weiyun
中科院分区：
文献类型：
--
作者：
Dong, Muchen;Di, Guohu;Shi, Weiyun
目的。探讨结膜下注射贝伐单抗对小鼠角膜神经、敏感性及上皮伤口愈合的影响。成年 C57BL/6 小鼠结膜下注射 1、2、5 或 25 mg/mL 贝伐珠单抗。采用整体抗-β3-微管蛋白荧光染色观察角膜神经。使用 Cochet-Bonnet 感觉计测量角膜敏感性。通过ELISA测定色素上皮衍生因子(PEDF)、神经生长因子(NGF)、胶质源性神经营养因子(GDNF)和睫状神经营养因子(CNTF)的蛋白水平。通过荧光素染色评价角膜上皮伤口愈合率。通过共同注射 PEDF、NGF 或 CNTF 来评估注射贝伐单抗后受损的小鼠角膜神经支配的恢复和上皮伤口愈合率。结果。结膜下注射贝伐单抗可引起正常小鼠明显的角膜神经变性、角膜敏感性减弱、角膜上皮伤口愈合和神经再生延迟，且随贝伐单抗注射浓度和次数的增加而更为显着。然而，单次注射1至5 mg/mL贝伐珠单抗的小鼠，角膜神经和敏感性逐渐改善和恢复。此外，贝伐单抗注射显着降低了角膜PEDF、NGF和CNTF含量，而外源性PEDF、NGF或CNTF补充剂减轻了结膜下注射贝伐单抗后对角膜神经、敏感性和上皮伤口愈合的损害。结论。结膜下注射贝伐单抗会损害正常小鼠的角膜神经支配、上皮伤口愈合和神经再生，这可能是由于角膜中神经营养因子含量减少所致。
PURPOSE. To investigate the effects of subconjunctival bevacizumab injection on the corneal nerve, sensitivity, and epithelial wound healing in mice.METHODS. Adult C57BL/6 mice were treated with subconjunctival injection of 1, 2, 5, or 25 mg/mL bevacizumab. The corneal nerve was observed with whole-mount anti-beta 3-tubulin fluorescence staining. Corneal sensitivity was measured with a Cochet-Bonnet esthesiometer. The protein levels of pigment epithelium-derived factor (PEDF), nerve growth factor (NGF), glial-derived neurotrophic factor (GDNF), and ciliary neurotrophic factor (CNTF) were measured by ELISA. The corneal epithelial wound-healing rate was evaluated by fluorescein staining. The recovery of impaired mouse corneal innervations and epithelial wound-healing rate following bevacizumab injection was evaluated with the co-injection of PEDF, NGF, or CNTF.RESULTS. Subconjunctival bevacizumab injection caused apparent corneal nerve degeneration, attenuated corneal sensitivity, and delayed corneal epithelial wound healing and nerve regeneration in normal mice, which was more significant with increased concentration and times of the bevacizumab injection. However, the corneal nerve and sensitivity gradually improved and recovered in mice with a single injection of 1 to 5 mg/mL bevacizumab. Moreover, the bevacizumab injection significantly decreased the corneal PEDF, NGF, and CNTF content, whereas exogenous PEDF, NGF, or CNTF supplement attenuated impairment of the corneal nerve, sensitivity, and epithelial wound healing after subconjunctival bevacizumab injection.CONCLUSIONS. Subconjunctival bevacizumab injection impairs corneal innervations, epithelial wound healing, and nerve regeneration in normal mice, which may be caused by the reduction of neurotrophic factor content in the cornea.