Role of short-wavelength blue light in the formation of cataracts and the expression of caspase-1, caspase-11, Gasdermin D in rat lens epithelial cells: insights into a novel pathogenic mechanism of cataracts

Abstract Backgroud: To examine the effects of short-wavelength blue light (SWBL) on cultured human lens epithelial cells (hLECs). The nosogenesis of cataracts after SWBL exposure was discussed. Methods: HLE-B3 hLECs were divided into 3 groups randomly: A: normal control group, which consisted of hLECs cultured in the dark; B: the caspase-1 inhibitor group; and C: the SWBL exposure group. After the SWBL (2500 lux) irradiation (for 8, 16, 24, and 32 h), the caspase-1 and gasdermin D (GSDMD) expression levels in HLE-B3 hLECs were examined using ELISA, immunofluorescence, and Western blotting analyses. Double-positive staining of HLE-B3 hLECs for activated and inhibited caspase-1 was used to confirm pyroptosis in hLECs by flow cytometry. Results: SWBL can cause cell death in HLE-B3 hLECs, but a caspase-1 inhibitor suppressed cell death. The flow cytometry results also confirmed the does-dependent of short-wavelength blue light irradiation on pyroptotic death of hLECs. Caspase-1 and GSDMD expression levels of all hLECs groups changed with short-wavelength blue light exposure times (8, 16, 24, and 32 h) and were higher in groups B and C than group A. The immunofluorescence results demonstrated that the expression of GSDMD-N was higher in the cell membrane in both the B and C groups than in the A group.Conclusion: The data indicate that SWBL induces pyroptotic programmed cell death by activation of the GSDMD signalling axis in HLE-B3 hLECs. These results provide new insights into the exploitation of new candidates for the prevention of cataracts.

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Role of Short-wavelength Blue Light in the Formation of Cataract and Expression of Caspase-1, -11 and Gasdermin D in rat Lens Epithelium Cells: Insight into the Novel Pathogenesis of Cataract.

10.21203/rs.2.18560/v1 ◽

2019 ◽

Author(s):

Yamin Wang ◽

Min Zhang ◽

Ying Sun ◽

Xiaohui Wang ◽

Zhaowei Song ◽

...

Keyword(s):

Blue Light ◽

Short Wavelength ◽

Light Exposure ◽

The Novel ◽

Blue Led ◽

Caspase 1 ◽

Epithelium Cells ◽

Experimental Group ◽

Insight Into

Abstract Background Cataracts have been verified to be associated with a number of risk factors. The sun and artificial light sources, including light-emitting diode (LED) and fluorescent light tubes, are the primary sources of short-wavelength blue light. With the increasing popularity of blue-rich LED-backlit display devices, our eyes are now exposed to more short-wavelength blue light than they were in the past. The goal of this study was to evaluate the role of short-wavelength blue light in the formation of cataract. Additionally, the pathogenesis of cataracts after short-wavelength light exposure was investigated.Methods SD rats were randomly divided into 2 main groups: a control group (10 rats each for the 4-, 8-, and 12-week groups) and an experimental group (10 rats each for the 4-, 8-, and 12-week groups). The rats in the experimental group were exposed to a short-wavelength blue LED lamp for 12 hours per day. After exposure to the blue LED lamp, the rats were maintained in total darkness for 12 hours, after which a 12-hour light/dark cycle was resumed. The intensity of the lamp was 3000 lux. At the end of the short-wavelength blue LED lamp exposure (for 4, 8, and 12 weeks), the expression levels of caspase-1, caspase-11 and gasdermin D (GSDMD) in rat epithelium cells (LECs) were examined in rat epithelial cells (LECs) using qRT-PCR and Western blotting analyses. Results After 6 weeks, cataracts had developed in the experimental rats (4/20 eyes). The clarity of the lens then gradually worsened with the duration of exposure. Twelve weeks later, all of the rat eyes had developed cataracts. Then the expression levels of caspase-1, caspase-11 and GSDMD at 4, 8, and 12 weeks were significantly higher in samples from rats exposed to a short-wavelength blue LED lamp than samples from control rat (p˂0.05). Conclusion The data indicate that pyroptosis play a key role of in cataracts induced by short-wavelength blue light exposure, highlighting caspase-1, caspase-11 and GSDMD as possible therapeutic targets for cataract treatment. This study might provide new insight into the novel pathogenesis of cataracts.

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Short-wavelength blue light contributes to the pyroptosis of human lens epithelial cells (hLECs)

Experimental Eye Research ◽

10.1016/j.exer.2021.108786 ◽

2021 ◽

pp. 108786

Author(s):

Xiaohui Wang ◽

Zhaowei Song ◽

Huazhang Li ◽

Kexin Liu ◽

Ying Sun ◽

...

Keyword(s):

Epithelial Cells ◽

Blue Light ◽

Short Wavelength ◽

Human Lens ◽

Lens Epithelial Cells ◽

Human Lens Epithelial Cells

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Ultrastructure changes and expression of NLRP3 inflammasome in lens anterior capsule of patient with cataracts associated with uveitis

10.21203/rs.2.16181/v1 ◽

2019 ◽

Author(s):

Chu Zhang ◽

Chong-Hui Ying ◽

siqin Sun ◽

yuechun Wen ◽

Zicheng Zhu

Keyword(s):

Epithelial Cells ◽

Ultrastructural Changes ◽

Lens Epithelial Cells ◽

Control Group ◽

Anterior Capsule ◽

Caspase 1 ◽

Pyrin Domain ◽

Age Related ◽

The Difference ◽

Domain 3

Abstract ● AIM : T o investigate the expression of nod-like receptor pyrin domain 3 (NLRP3) in lens anterior capsule of Uveitis associated with cataract and observe the ultrastructural changes of them . ● Methods: 17(22 eyes) cases of uveitis associated with cataract were selected a s experimental group and 1 0 (18 eyes) cases of age-related cataract were selected a s contro l group. The expressions of NLRP3, apoptosis-related speckle protein （ASC） and caspase-1 protein were tested by immunohistochemical and the ultrastructural changes of anterior capsul e was observed under electron microscope. ● Results: The expression of NLRP3 、 caspase-1 and ASC in the anterior capsu le of cataracts associated with uveitis was significantly higher than that of the control group, the difference was statistically significant (p ＜ 0.05). The apoptotic changes of lens epithelial cells in uveitis associated with cataract were obvious, and the apoptotic changes of lens epithelial cells were mild in age-related cataract patients. ● Conclusion: Strongly postive expressed NLRP3 inflamma some and obvious apoptotic changes are founded in the lens epithelial cells of patients with uveitis associat ed with cataract, suggesting that NLRP3 inflamma some and the apoptosis of lens epithelial cells may play a role in the progress of uveitis associat ed with cataract.

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Thioredoxin Binding Protein-2 Regulates Autophagy of Human Lens Epithelial Cells under Oxidative Stress via Inhibition of Akt Phosphorylation

Oxidative Medicine and Cellular Longevity ◽

10.1155/2016/4856431 ◽

2016 ◽

Vol 2016 ◽

pp. 1-17 ◽

Cited By ~ 5

Author(s):

Jiaojie Zhou ◽

Ke Yao ◽

Yidong Zhang ◽

Guangdi Chen ◽

Kairan Lai ◽

...

Keyword(s):

Oxidative Stress ◽

Epithelial Cells ◽

Cell Viability ◽

Binding Protein ◽

Negative Regulator ◽

Human Lens ◽

Lens Epithelial Cells ◽

Human Lens Epithelial Cells ◽

Age Related

Oxidative stress plays an essential role in the development of age-related cataract. Thioredoxin binding protein-2 (TBP-2) is a negative regulator of thioredoxin (Trx), which deteriorates cellular antioxidant system. Our study focused on the autophagy-regulating effect of TBP-2 under oxidative stress in human lens epithelial cells (LECs). Human lens epithelial cells were used for cell culture and treatment. Lentiviral-based transfection system was used for overexpression of TBP-2. Cytotoxicity assay, western blot analysis, GFP/mCherry-fused LC3 plasmid, immunofluorescence, and transmission electronic microscopy were performed. The results showed that autophagic response of LECs with increased LC3-II, p62, and GFP/mCherry-LC3 puncta (P<0.01) was induced by oxidative stress. Overexpression of TBP-2 further strengthens this response and worsens the cell viability (P<0.01). Knockdown of TBP-2 attenuates the autophagic response and cell viability loss induced by oxidative stress. TBP-2 mainly regulates autophagy in the initiation stage, which is mTOR-independent and probably caused by the dephosphorylation of Akt under oxidative stress. These findings suggest a novel role of TBP-2 in human LECs under oxidative stress. Oxidative stress can cause cell injury and autophagy in LECs, and TBP-2 regulates this response. Hence, this study provides evidence regarding the role of TBP-2 in lens and the possible mechanism of cataract development.

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Long Non-Coding RNA H19 Regulates Human Lens Epithelial Cells Function

Cellular Physiology and Biochemistry ◽

10.1159/000494003 ◽

2018 ◽

Vol 50 (1) ◽

pp. 246-260 ◽

Cited By ~ 6

Author(s):

Xin Liu ◽

Chang Liu ◽

Kun Shan ◽

Shujie Zhang ◽

Yi Lu ◽

...

Keyword(s):

Oxidative Stress ◽

Epithelial Cells ◽

Differentially Expressed ◽

Human Lens ◽

Lens Epithelial Cells ◽

Double Staining ◽

Human Lens Epithelial Cells ◽

And Migration ◽

Proliferation And Migration

Background/Aims: Age-related cataract (ARC) remains the leading cause of visual impairment among the elderly population. Long non-coding RNAs (lncRNAs) have emerged as potential regulators in many ocular diseases. However, the role of lncRNAs in nuclear ARC, a subtype of ARC, requires further elucidation. Methods: LncRNA sequencing was performed to identify differentially expressed lncRNAs between the capsules of transparent and nuclear ARC lenses. Expression validation was confirmed by qRT-PCR. MTT assay, Calcein-AM and propidium iodide double staining, Rhodamine 123 and Hoechst double staining, EdU and transwell assay were used to determine the role of H19 or miR-675 in the viability, apoptosis, proliferation and migration of primary cultured human lens epithelial cells (HLECs). Bioinformatics and luciferase reporter assays were used to identify the binding target of miR-675. Results: Sixty-three lncRNAs are differentially expressed between the capsules of transparent and nuclear ARC lenses. One top abundantly expressed lncRNA, H19, is significantly up-regulated in the nuclear ARC lens capsules and positively associated with nuclear ARC grade. H19 knockdown accelerates apoptosis development and reduces the proliferation and migration of HLECs upon oxidative stress. H19 is the precursor of miR-675, and a reduction of H19 inhibits miR-675 expression. miR-675 regulates CRYAA expression by targeting the binding site within the 3’UTR. Moreover, miR-675 increases the proliferation and migration while decreasing the apoptosis of HLECs upon oxidative stress. Conclusion: H19 regulates HLECs function through miR-675-mediated CRYAA expression. This finding would provide a novel insight into the pathogenesis of nuclear ARC.

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Response of Lens Epithelial Cells to Injury: Role of Lumican in Epithelial-Mesenchymal Transition

Investigative Opthalmology & Visual Science ◽

10.1167/iovs.02-1059 ◽

2003 ◽

Vol 44 (5) ◽

pp. 2094 ◽

Cited By ~ 78

Author(s):

Shizuya Saika ◽

Takeshi Miyamoto ◽

Sai-ichi Tanaka ◽

Takeshi Tanaka ◽

Iku Ishida ◽

...

Keyword(s):

Epithelial Cells ◽

Epithelial Mesenchymal Transition ◽

Lens Epithelial Cells ◽

Mesenchymal Transition

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The role of melatonin as an antioxidant in human lens epithelial cells

Free Radical Research ◽

10.3109/10715762.2013.808743 ◽

2013 ◽

Vol 47 (8) ◽

pp. 635-642 ◽

Cited By ~ 26

Author(s):

J. Bai ◽

L. Dong ◽

Z. Song ◽

H. Ge ◽

X. Cai ◽

...

Keyword(s):

Epithelial Cells ◽

Human Lens ◽

Lens Epithelial Cells ◽

Human Lens Epithelial Cells

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The Role of Nuclear Factor of Activated T Cells 5 in Hyperosmotic Stress-Exposed Human Lens Epithelial Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22126296 ◽

2021 ◽

Vol 22 (12) ◽

pp. 6296

Author(s):

Gyu-Nam Kim ◽

Young-Sool Hah ◽

Hyemin Seong ◽

Woong-Sun Yoo ◽

Mee-Young Choi ◽

...

Keyword(s):

T Cells ◽

Epithelial Cells ◽

Hyperosmotic Stress ◽

Human Lens ◽

Lens Epithelial Cells ◽

Nuclear Factor ◽

Activated T Cells ◽

Interacting Protein ◽

Human Lens Epithelial Cells

We investigated the role of nuclear factor of activated T cells 5 (NFAT5) under hyperosmotic conditions in human lens epithelial cells (HLECs). Hyperosmotic stress decreased the viability of human lens epithelial B-3 cells and significantly increased NFAT5 expression. Hyperosmotic stress-induced cell death occurred to a greater extent in NFAT5-knockout (KO) cells than in NFAT5 wild-type (NFAT5 WT) cells. Bcl-2 and Bcl-xl expression was down-regulated in NFAT5 WT cells and NFAT5 KO cells under hyperosmotic stress. Pre-treatment with a necroptosis inhibitor (necrostatin-1) significantly blocked hyperosmotic stress-induced death of NFAT5 KO cells, but not of NFAT5 WT cells. The phosphorylation levels of receptor-interacting protein kinase 1 (RIP1) and RIP3, which indicate the occurrence of necroptosis, were up-regulated in NFAT5 KO cells, suggesting that death of these cells is predominantly related to the necroptosis pathway. This finding is the first to report that necroptosis occurs when lens epithelial cells are exposed to hyperosmolar conditions, and that NFAT5 is involved in this process.

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