Kang Li-Hua, Zhang Su, Jiang Sheng, Hu Nan
Eye Institute, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
Nanjing Medical University Affiliated Eye Hospital, Nanjing 210000, Jiangsu Province, China.
Int J Ophthalmol. 2019 Sep 18;12(9):1395-1401. doi: 10.18240/ijo.2019.09.04. eCollection 2019.
To investigate the activation of autophagy in rat retina after optic nerve crush (ONC) and evaluate its relationship with apoptosis of retinal ganglion cells (RGCs).
The ONC model was established. Western blots were performed to investigate expression of p62, LC3 and Beclin-1. Transmission electron microscopy was performed to discover the autophagosomes in the retina after ONC. Immunohistochemistry was used to confirm the distribution of LC3. TUNEL was performed to confirm the relationship between autophagy and RGC apoptosis.
p62/Beclin-1 ratio was declined shortly after ONC until to day 7 after ONC and then restored to a normal level at day 21. There was an opposite change in the LC3-II/LC3I ratio in the retina compared to the p62/Beclin-1 ratio. Increased autophagosomes were found after ONC using transmission electron microscopy, and most of the LC3-stained cells were colocalized with RGCs and Müller cells. More LC3-immunoreactive cells and apoptotic RGCs were found on day 7 following ONC.
Possible activation of autophagy in RGCs after ONC; autophagy mainly occurred in RGCs and Müller cells, and the apoptosis of RGCs after ONC may be partly associated with autophagic activation.
研究视神经挤压伤(ONC)后大鼠视网膜自噬的激活情况,并评估其与视网膜神经节细胞(RGCs)凋亡的关系。
建立ONC模型。采用蛋白质免疫印迹法检测p62、LC3和Beclin-1的表达。采用透射电子显微镜观察ONC后视网膜中的自噬体。采用免疫组织化学法确定LC3的分布。采用TUNEL法确定自噬与RGC凋亡之间的关系。
ONC后不久,p62/Beclin-1比值下降,直至ONC后第7天,然后在第21天恢复到正常水平。视网膜中LC3-II/LC3I比值与p62/Beclin-1比值呈相反变化。采用透射电子显微镜观察发现ONC后自噬体增加,且大多数LC3染色细胞与RGCs和Müller细胞共定位。ONC后第7天发现更多LC3免疫反应性细胞和凋亡RGCs。
ONC后RGCs中可能存在自噬激活;自噬主要发生在RGCs和Müller细胞中,ONC后RGCs的凋亡可能部分与自噬激活有关。