Eye Hospital, The First Affiliated Hospital of Harbin Medical University, China.
Eye Hospital, The First Affiliated Hospital of Harbin Medical University, China; Department of Ophthalmology, Second Hospital of Heilongjiang Province, Harbin, China.
Exp Eye Res. 2021 Nov;212:108786. doi: 10.1016/j.exer.2021.108786. Epub 2021 Oct 5.
The purpose of this study is to examine the effect of short-wavelength blue light (SWBL) on cultured human lens epithelial cells (hLECs). The pathogenesis of cataracts after SWBL exposure is discussed.
HLE-B3 hLECs were randomly divided into 3 groups: the NC group, which was grown in a dark incubator; the acetyl (Ac)-Tyr-Val-Ala-Asp-chloromethyl ketone (AC-YVAD-CMK) treatment group; and the SWBL exposure group. After SWBL (2500 lux) irradiation (for 8, 16, 24, and 32 h), caspase-1 and gasdermin D (GSDMD) expression levels in HLE-B3 hLECs were examined using ELISA, immunofluorescence staining, and Western blotting analyses. Double-positive staining of hLECs for activated and inhibited caspase-1 was used to determine pyroptosis in HLE-B3 hLECs.
SWBL led to hLEC death, but a caspase-1 inhibitor suppressed cell death. The flow cytometry results also confirmed the dose-dependent effect of SWBL irradiation on the pyroptotic death of hLECs. Caspase-1 and GSDMD expression levels in all hLEC groups changed with blue light exposure times (8, 16, 24, and 32 h) and were higher in the AC-YVAD-CMK and SWBL exposure groups than in the NC group. The immunofluorescence results revealed higher GSDMD-N expression in the cell membrane of both the AC-YVAD-CMK and SWBL exposure groups than in the NC group.
Based on the data, SWBL induces pyroptotic programmed cell death by activating the GSDMD signalling axis in HLE-B3 hLECs. These results provide new insights into the exploitation of new candidates for the prevention of cataracts.
本研究旨在探讨短波长蓝光（SWBL）对培养的人晶状体上皮细胞（hLECs）的影响。讨论了 SWBL 暴露后白内障的发病机制。
将 HLE-B3 hLECs 随机分为 3 组：NC 组，在黑暗孵育箱中生长；乙酰（Ac）-Tyr-Val-Ala-Asp-氯甲基酮（AC-YVAD-CMK）处理组；以及 SWBL 暴露组。SWBL（2500 lux）照射（8、16、24 和 32 h）后，采用 ELISA、免疫荧光染色和 Western blot 分析检测 HLE-B3 hLECs 中 caspase-1 和 gasdermin D（GSDMD）的表达水平。用激活和抑制 caspase-1 的 hLECs 双重阳性染色来确定 HLE-B3 hLECs 中的细胞焦亡。
SWBL 导致 hLEC 死亡，但 caspase-1 抑制剂抑制了细胞死亡。流式细胞术结果也证实了 SWBL 照射对 hLEC 细胞焦亡的剂量依赖性影响。所有 hLEC 组的 caspase-1 和 GSDMD 表达水平随蓝光照射时间（8、16、24 和 32 h）而变化，且 AC-YVAD-CMK 和 SWBL 暴露组的表达水平均高于 NC 组。免疫荧光结果显示，AC-YVAD-CMK 和 SWBL 暴露组的细胞膜上 GSDMD-N 的表达均高于 NC 组。
基于这些数据，SWBL 通过激活 GSDMD 信号通路诱导 HLE-B3 hLECs 发生细胞焦亡程序性细胞死亡。这些结果为开发新的白内障预防候选物提供了新的思路。
