Impaired Autophagy Causes Severe Corneal Neovascularization

作者全名："Yi, Kun; Yang, Yuping; Yuan, Ye; Xiang, Yingqian; Zhou, Shanbi"

作者地址："[Yi, Kun; Yang, Yuping; Yuan, Ye; Xiang, Yingqian; Zhou, Shanbi] Chongqing Med Univ, Affiliated Hosp 1, Chongqing Key Lab Ophthalmol, Chongqing 400016, Peoples R China; [Yi, Kun; Yang, Yuping; Yuan, Ye; Xiang, Yingqian; Zhou, Shanbi] Chongqing Med Univ, Affiliated Hosp 1, Chongqing Eye Inst, Chongqing 400016, Peoples R China"

通信作者："Zhou, SB (通讯作者)，Chongqing Med Univ, Affiliated Hosp 1, Chongqing Key Lab Ophthalmol, Chongqing 400016, Peoples R China.; Zhou, SB (通讯作者)，Chongqing Med Univ, Affiliated Hosp 1, Chongqing Eye Inst, Chongqing 400016, Peoples R China."

来源：CELLS

ESI学科分类：BIOLOGY & BIOCHEMISTRY

WOS号：WOS:000897271700001

JCR分区：Q2

影响因子：6

年份：2022

卷号：11

期号：23

开始页：　

结束页：　

文献类型：Article

关键词：corneal neovascularization; autophagy; macrophage polarization; alkali burn

摘要："Purpose: To investigate the role of macrophage autophagy in the process of corneal neovascularization (CNV). Methods: In vivo, mice CNV was induced by alkali injury and compared with rapamycin-treated alkaline burn mice. Western blot was used to determine the autophagic status of the macrophages. We quantified the levels of macrophage polarization markers (CD86, INOS, CD163, CD206) by RT-qPCR and measured inflammatory factors through ELISA (IL-6 and TNF-alpha) in the early phase after injury. In vitro, the human umbilical vein endothelial cells (HUVECs) were co-cultured with macrophage-conditioned medium (MCM) induced by the THP-1 cell line to simulate the neovascular microenvironment. The vascularization capacity of HUVECs was examined using the CCK-8 assay kit, tube formation assay, and scratch wound-healing assay. Results: In vivo, the mRNA expression of Beclin-1 and ATG5 was increased, together with the upregulation of M1 macrophage markers (CD86 and INOS) in corneas after early alkali injury. The area of CNV is effectively relieved in the rapamycin-treated mice. In vitro, upregulation of autophagy level by pretreatment with 3-methyladenine (3-MA) could increase the mRNA expression of the M1 markers. Macrophage-conditioned medium with impaired autophagy contains more IL-6 and TNF-alpha compared to the M1 macrophage-conditioned medium, promoting HUVEC proliferation, migration, and tube formation capacity. Enhancing the autophagy level with rapamycin (RAPA) could reverse this phenomenon. Conclusions: Impaired autophagy promoted macrophage polarization toward M1 type and increased the expression of IL-6 and TNF-alpha, which led to severe CNV. Using the autophagy activator (RAPA) could effectively alleviate CNV by promoting autophagy."

基金机构：Key Generic Technologies for Third-Party Services Based on Health Management (CSTC) [2015shmh-ztzx10006]; Chongqing High-tech Zone Application Study of Domestic in Vitro Diagnostic Medical Devices (CSCT) [2020jscx-sbqwX0004]; Chongqing Key Laboratory of Ophthalmology (CSTC) [2008CA5003]

基金资助正文："This study was supported by Key Generic Technologies for Third-Party Services Based on Health Management (CSTC, 2015shmh-ztzx10006), Chongqing High-tech Zone Application Study of Domestic in Vitro Diagnostic Medical Devices (CSCT, 2020jscx-sbqwX0004), and the Chongqing Key Laboratory of Ophthalmology (CSTC, 2008CA5003)."