免疫印迹
荧光素
分子生物学
吞噬作用
逆转录聚合酶链式反应
染色
免疫学
炎症
男科
生物
化学
基因表达
病理
医学
生物化学
物理
量子力学
基因
荧光
作者
Sasa Zhang,Xing Lü,Cui Li,Qian Wang,Shanshan Yang,Xudong Peng,Liting Hu,Guiqiu Zhao,Jing Lin
标识
DOI:10.1080/02713683.2023.2262169
摘要
To confirm the expression and investigate the role of LC3-associated phagocytosis (LAP) in dry eye disease (DED).The DED model of mice was established by scopolamine subcutaneous injection in a low-humidity environment chamber. Tear secretion test and corneal fluorescein sodium staining were used to evaluate the severity of DED. Expression levels of Rubicon, microtubule-associated protein light chain 3-II (LC3-II), Beclin-1 and autophagy-related gene-7 (Atg-7) in corneas of mice with DED were tested by western blot. Cell Counting Kit-8 (CCK-8) assay was used to detect the effects of different concentrations of hypertonic solutions on the proliferation activity of human corneal epithelial cells (HCECs). The expression levels of Dectin-1, IL-6 and IL-1β in HCECs after stimulation with different concentrations of hypertonic solutions were tested. The expressions of Rubicon, LC3-II, Beclin-1 and ATG-7 in HCECs were detected by reverse transcription polymerase chain reaction (RT-PCR). After being pretreated with 10 μM si-Rubicon, the severity of the disease was documented by corneal fluorescein sodium staining. And the expression levels of IL-6 and IL-1β were also tested by RT-PCR.Compared with the normal control group, the corneal fluorescein sodium staining scores and tear secretion were significantly reduced. Rubicon, LC3-II, Beclin-1 and ATG-7 were significantly elevated. CCK-8 showed that the 400 and 450 mOsM hypertonic solutions did not affect the proliferation activity of HCECs. The expression of Dectin-1, IL-1β and IL-6 were elevated after stimulation with 450 mOsM solution. LC3-II, Rubicon, ATG-7 and Beclin-1 increased after stimulation with 450 mOsM hyperosmolar solution in HCECs. Corneal fluorescein staining showed that si-Rubicon increased the severity of DED in mice. Moreover, the mRNA expressions of inflammatory factors IL-1β and IL-6 in the cornea of mice were significantly increased.DED increased the expression of proteins associated with LAP. LAP could play an anti-inflammatory effect in DED.
科研通智能强力驱动
Strongly Powered by AbleSci AI