Dong Muchen, Di Guohu, Zhang Xiaoping, Zhou Qingjun, Shi Weiyun
School of Medicine and Life Sciences, University of Jinan-Shandong Academy of Medical Sciences, Jinan, China 2Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of Medical Sciences, Qingdao, China.
Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of Medical Sciences, Qingdao, China.
Invest Ophthalmol Vis Sci. 2017 Mar 1;58(3):1469-1477. doi: 10.1167/iovs.16-20926.
To investigate the effects of subconjunctival bevacizumab injection on the corneal nerve, sensitivity, and epithelial wound healing in mice.
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-β3-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.
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.
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.
研究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。
对成年C57BL/6小鼠进行结膜下注射1、2、5或25mg/mL贝伐单抗治疗。采用全层抗β3-微管蛋白荧光染色观察角膜神经。用Cochet-Bonnet感觉计测量角膜敏感性。通过酶联免疫吸附测定法测量色素上皮衍生因子（PEDF）、神经生长因子（NGF）、胶质细胞源性神经营养因子（GDNF）和睫状神经营养因子（CNTF）的蛋白水平。通过荧光素染色评估角膜上皮伤口愈合率。通过联合注射PEDF、NGF或CNTF评估贝伐单抗注射后受损小鼠角膜神经支配的恢复情况及上皮伤口愈合率。
结膜下注射贝伐单抗导致正常小鼠出现明显的角膜神经变性、角膜敏感性减弱以及角膜上皮伤口愈合和神经再生延迟，且随着贝伐单抗注射浓度和次数的增加，这种情况更为显著。然而，单次注射1至5mg/mL贝伐单抗的小鼠角膜神经和敏感性逐渐改善并恢复。此外，贝伐单抗注射显著降低了角膜PEDF、NGF和CNTF含量，而外源性补充PEDF、NGF或CNTF可减轻结膜下注射贝伐单抗后角膜神经、敏感性及上皮伤口愈合的损伤。
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降低所致。
