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Updated: Oct 1, 2025
Induction of Ocular Surface Inflammation and Collection of Involved Tissues
Published on: August 4, 2022
Necroptosis contributes to airborne particulate matter-induced ocular surface injury
Kexin Shi1, Qichuan Yin1, Xiajing Tang1
1The Eye Center, Second Affiliated Hospital of School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China; Zhejiang Provincial Key Lab of Ophthalmology, Hangzhou, Zhejiang Province, China.
Abstract:
In this study, we explored the role of necroptosis in the pathogenesis of ocular surface injury caused by airborne particulate matter (PM). Human corneal epithelial (HCE) cells and mouse ocular surface were treated with PM exposure and compared with non-exposed groups. The expression of necroptosis-related proteins was measured by immunoblotting in HCE cell groups. Cell damages were detected using CCK-8, flow cytometry, and immunofluorescence staining. In the mouse model, hematoxylin and eosin (H&E) staining and corneal fluorescein sodium staining were assessed. In addition, the expression of inflammatory cytokines and mucin were examined via Enzyme-linked immunosorbent assay (ELISA), immunofluorescence staining and/or quantitative RT -PCR (qRT-PCR), both in vitro and in vivo. Our research showed that PM exposure may trigger HCE cell damage via necroptosis. Necrostatin-1(Nec-1), one of the specific inhibitors of necroptosis, can markedly reduce PM-induced HCE cell damage. HCE cell damage markers included decreased cell viability, increased intracellular reactive oxygen species (ROS) levels, and loss of mitochondrial membrane potential. At the same time, Nec-1 inhibited the increased inflammatory cytokines and the decreased mucin expression caused by PM exposure in HCE cells. Nec-1 also reduced corneal inflammation and mucin underproduction in mouse ocular surface after PM exposure. Our study demonstrated that necroptosis is involved in the pathogenesis of PM exposure-related ocular surface injury, including inflammation and insufficient mucin production in the cornea, which can be rescued by inhibitor Nec-1. This suggests Nec-1 could be a novel therapeutic target for ocular surface disorders, especially dry eye disease, which is caused by the exacerbation of airborne PM pollution.
Insights
Airborne particulate matter (PM) causes ocular surface injury by triggering necroptosis, a programmed cell death. The necroptosis inhibitor Necrostatin-1 (Nec-1) effectively reduced PM-induced eye damage and inflammation.
Area of Science:
- Ophthalmology
- Cell Biology
- Environmental Health
Background:
- Airborne particulate matter (PM) poses a significant threat to ocular surface health.
- The precise mechanisms underlying PM-induced ocular surface injury are not fully understood.
- Necroptosis, a regulated form of necrosis, is increasingly recognized in various disease pathologies.
Purpose of the Study:
- To investigate the role of necroptosis in the pathogenesis of ocular surface injury induced by airborne particulate matter (PM).
- To evaluate the therapeutic potential of necroptosis inhibition in mitigating PM-induced ocular surface damage.
Main Methods:
- Human corneal epithelial (HCE) cells and a mouse ocular surface model were exposed to PM.
- Necroptosis-related protein expression, cell viability, reactive oxygen species (ROS) levels, and mitochondrial membrane potential were assessed.
- Inflammatory cytokine and mucin expression were quantified using ELISA, immunofluorescence, and qRT-PCR.
- The effects of Necrostatin-1 (Nec-1), a necroptosis inhibitor, were evaluated both in vitro and in vivo.
Main Results:
- PM exposure induced HCE cell damage, characterized by decreased viability, increased ROS, and loss of mitochondrial membrane potential, via necroptosis.
- Nec-1 significantly attenuated PM-induced HCE cell damage and reduced inflammatory cytokine production.
- Nec-1 inhibited PM-induced decrease in mucin expression in HCE cells.
- In vivo, Nec-1 treatment reduced corneal inflammation and mucin underproduction in the mouse ocular surface following PM exposure.
Conclusions:
- Necroptosis plays a critical role in the pathogenesis of PM-induced ocular surface injury.
- Nec-1 demonstrates significant therapeutic potential in ameliorating PM-related ocular surface inflammation and dysfunction.
- Targeting necroptosis with inhibitors like Nec-1 offers a promising strategy for treating ocular surface disorders, including dry eye disease exacerbated by air pollution.
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