Department of Ophthalmology, The First Affiliated Hospital of Fujian Medical University, Fuzhou, China.
Department of Ophthalmology, The First Affiliated Hospital of Fujian Medical University, Fuzhou, China.
Clinics (Sao Paulo). 2023 Jul 4;78:100241. doi: 10.1016/j.clinsp.2023.100241. eCollection 2023.
To explore the mechanism underlying Müller Cell Pyroptosis (MCP) and its role in the development of Proliferative Vitreoretinopathy (PVR).
The expression of pyroptosis-related factors, namely, cysteinyl aspartate-specific proteinase (caspase-1), interleukin (IL)-1β, IL-18, and Gasdermin D (GSDMD), was detected by quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) and western blotting at the mRNA and protein levels, respectively, in retinal tissues. Müller and spontaneously Arising Retinal Pigment Epithelia (ARPE)-19 primary cells with GSDMD overexpression or knockdown were cultivated. Western blotting was used to detect the levels of the following pyroptosis-related factors in retinal tissues: caspase-1, IL-1β, IL-18, and GSDMD. Through Cell Adhesion (CA) experiments, the changes in ARPE-19 CA in each group were observed. The migration and invasion of ARPE-19 cells were measured using the Transwell assay. The proliferation of ARPE-19 cells was measured with a Cell Counting Kit 8 (CCK-8) assay. Finally, the expression of the cytokines IL-1β and IL-18 in the ARPE-19 cell culture medium was detected using the Enzyme-Linked Immunosorbent Assay (ELISA).
Compared with the surrounding normal tissues, the expression of caspase-1, IL-1β, IL-18, and GSDMD at the protein and mRNA levels in the retinal proliferative membrane samples of the patients decreased significantly (p < 0.05). MCP significantly enhanced ARPE-19 CA, migration and invasion, proliferation, and cytokine expression (p < 0.05).
MCP can promote the development of PVR lesions.
探讨 Müller 细胞细胞焦亡（MCP）的发生机制及其在增生性玻璃体视网膜病变（PVR）发展中的作用。
通过定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法，分别检测视网膜组织中细胞焦亡相关因子胱天蛋白酶-1（caspase-1）、白细胞介素（IL）-1β、IL-18 和 Gasdermin D（GSDMD）的表达水平。培养过表达或敲低 GSDMD 的 Müller 细胞和自发出现的视网膜色素上皮细胞（ARPE）-19 原代细胞。Western blot 检测各组视网膜组织中焦亡相关因子 caspase-1、IL-1β、IL-18 和 GSDMD 的水平。通过细胞黏附（CA）实验观察各组 ARPE-19 CA 的变化。通过 Transwell 实验测量 ARPE-19 细胞的迁移和侵袭。用细胞计数试剂盒 8（CCK-8）法测量 ARPE-19 细胞的增殖。最后，用酶联免疫吸附试验（ELISA）检测 ARPE-19 细胞培养上清液中细胞因子 IL-1β和 IL-18 的表达。
与周围正常组织相比，患者视网膜增生性膜样本中 caspase-1、IL-1β、IL-18 和 GSDMD 的蛋白和 mRNA 水平表达均显著降低（p<0.05）。MCP 明显增强了 ARPE-19 CA、迁移和侵袭、增殖和细胞因子表达（p<0.05）。
MCP 可促进 PVR 病变的发展。
