Short-Wavelength Blue Light Contributes to Pyroptosis of Human Lens Epithelial Cells (hLECs) by Caspase-1-GSDMD Signalling Axis Activation

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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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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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

BMC Ophthalmology ◽

10.1186/s12886-020-01565-z ◽

2020 ◽

Vol 20 (1) ◽

Author(s):

Yamin Wang ◽

Min Zhang ◽

Ying Sun ◽

Xiaohui Wang ◽

Zhaowei Song ◽

...

Keyword(s):

Epithelial Cells ◽

Blue Light ◽

Short Wavelength ◽

Pathogenic Mechanism ◽

Lens Epithelial Cells ◽

Caspase 1

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Cadmium-induced induction of cell death in human lens epithelial cells: Implications to smoking associated cataractogenesis

Toxicology Letters ◽

10.1016/j.toxlet.2010.04.021 ◽

2010 ◽

Vol 198 (1) ◽

pp. 56-62 ◽

Cited By ~ 33

Author(s):

Nilesh M. Kalariya ◽

Bindu Nair ◽

Denish K. Kalariya ◽

Nancy K. Wills ◽

Frederik J.G.M. van Kuijk

Keyword(s):

Cell Death ◽

Epithelial Cells ◽

Human Lens ◽

Lens Epithelial Cells ◽

Human Lens Epithelial Cells

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Coenzyme Q10 prevents human lens epithelial cells from light-induced apoptotic cell-death by reducing oxidative stress and stabilizing BAX/Bcl-2 ratio

Acta Ophthalmologica ◽

10.1111/j.1755-3768.2009.337.x ◽

2009 ◽

Vol 87 ◽

pp. 0-0

Author(s):

M KERNT ◽

C HIRNEISS ◽

AS NEUBAUER ◽

MW ULBIG ◽

A KAMPIK

Keyword(s):

Oxidative Stress ◽

Cell Death ◽

Epithelial Cells ◽

Coenzyme Q10 ◽

Apoptotic Cell ◽

Apoptotic Cell Death ◽

Human Lens ◽

Lens Epithelial Cells ◽

Human Lens Epithelial Cells

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The effect of Sep15 gene silencing on the expression of crystall oid protein in human lens epithelial cells

E3S Web of Conferences ◽

10.1051/e3sconf/202018503005 ◽

2020 ◽

Vol 185 ◽

pp. 03005

Author(s):

Meng Wang

Keyword(s):

Endoplasmic Reticulum ◽

Epithelial Cells ◽

Protein Expression ◽

Gene Silencing ◽

Human Lens ◽

Lens Epithelial Cells ◽

Lens Protein ◽

Expression Levels ◽

Human Lens Epithelial Cells ◽

Mrna And Protein Expression

Objective: To investigate the effect of Sep15 gene silencing on lens protein expression in human lens epithelial cells. Method: Human lens epithelial cell (HLEC) SRA01/04 was used as the study object, Using MTT method, RT-PCRs, Small molecule RNA interference technology and protein immunoblotting method, The survival rate of hLE cells stimulated by different concentrations of bFGF and Tm (tunicamycin, an endoplasmic reticulum stress inducing reagent), the changes of the mRNA and protein expression levels of intracellular beta-crystallin and GRP78 proteins, and the changes of Sep15 protein expression levels in hLE cells were determined, respectively, to evaluate the effect of Sep15 gene silencing on the expression of lens protein in human lens epithelial cells. Result: First, The survival rate of hLE cells induced by endoplasmic reticulum stress inducing reagents tunicamycin (Tm) and bFGF was investigated by MTT method. The results showed that a certain concentration of Tm and bFGF could inhibit or lead to the death of hLE cells; Secondly, The effects of Tm on the levels of mRNA and protein expression of the endoplasmic reticulum stress marker protein GRP78 and the effects of bFGF on the levels of alpha-, were detected by real-time fluorescence quantitative PCR and protein immunoblotting, respectively, Effects of beta-crystallin mRNA and protein expression levels, To determine the optimal concentration and time of Tm and bFGF on hLE cells, The results showed that 40 ng/mL of bFGF-treated cells for 48 h was the optimal reaction condition for bFGF to stimulate the differentiation of lens epithelial cells. Third, The co-action of Tm and bFGF on hLE cells was detected by real-time fluorescence quantitative PCR and protein immunoblotting. For Intracellular GRP78, α-, β- crystallin, Effects of mRNA and protein expression levels, The results showed that the occurrence of ER stress could upregulate its expression. Fourth, The changes of Sep15 protein expression level in hLE cells after Sep15 gene silencing and the effects of Tm and bFGF on Sep15 gene silencing in cells were detected by Western blotting. The results showed that the addition of Tm and bFGF basically did not affect the silencing effect of Sep15 gene. Fifth, The expression of GRP78 and β- crystallin in hLE cells after silencing of Sep15 gene was detected by Western blot. The results showed that Sep15 may be involved in the process of protecting the differentiation of lens epithelial cells. Conclusion: Sep15 gene silencing has an inhibitory effect on lens protein expression in human lens epithelial cells, and strengthening its clinical research is of positive significance for improving the clinical treatment and prevention of cataract diseases.

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Coenzyme Q10 prevents human lens epithelial cells from light-induced apoptotic cell death by reducing oxidative stress and stabilizing BAX / Bcl-2 ratio

Acta Ophthalmologica ◽

10.1111/j.1755-3768.2010.01875.x ◽

2010 ◽

Vol 88 (3) ◽

pp. e78-e86 ◽

Cited By ~ 16

Author(s):

Marcus Kernt ◽

Christoph Hirneiss ◽

Aljoscha S. Neubauer ◽

Michael W. Ulbig ◽

Anselm Kampik

Keyword(s):

Oxidative Stress ◽

Cell Death ◽

Epithelial Cells ◽

Coenzyme Q10 ◽

Apoptotic Cell ◽

Apoptotic Cell Death ◽

Human Lens ◽

Lens Epithelial Cells ◽

Human Lens Epithelial Cells

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Oxidized low-density lipoprotein enhances Orai3 expression levels to increase proliferation of human lens epithelial cells

10.21203/rs.3.rs-28318/v1 ◽

2020 ◽

Author(s):

Ru Zhang ◽

Suwen Bai ◽

Yang Fang ◽

Zhixin Zhou ◽

Linghui Chen ◽

...

Keyword(s):

Cell Proliferation ◽

Epithelial Cells ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Human Lens ◽

Lens Epithelial Cells ◽

Low Density ◽

Oxidized Low Density Lipoprotein ◽

Expression Levels ◽

Human Lens Epithelial Cells

Abstract Background Serum lipid levels, especially of oxidized low-density lipoprotein (oxLDL), are related to the development of cataracts, but the mechanisms underlying the role of oxLDL in cataract development in human lens epithelial cells (HLEpiCs) remain unclear. Calcium (Ca2+) overload is also known to be involved in lens turbidity. Store-operated Ca2+ entry (SOCE) is an important pathway mediating Ca2+ influx in lens epithelial cells. The Orai family proteins, which form a type of SOCE channel, localize at the plasma membrane and control Ca2+ influx in response to the depletion of Ca2+ stores in the endoplasmic reticulum. Methods In the present study, we used RNA sequencing, western blot analyses, cell proliferation assays, Ca2+ measurements, and small interfering (si)RNA transfections to investigate the effects of oxLDL on SOCE and proliferation of human lens epithelial cells (HLEpiCs). Results The key findings of our study that suggest this conclusion are that (1) oxLDL enhances the expression levels of Orai3, but not Orai1 or Orai2, in HLEpiCs in vitro; (2) oxLDL significantly increases the proliferation of HLEpiCs in a concentration-dependent manner; (3) oxLDL significantly increases ATP-induced SOCE without affecting Ca2+ release in HLEpiCs; and (4) knockdown of Orai3 significantly reduces cell proliferation and ATP-induced SOCE in HLEpiCs. Conclusions These findings suggest that Ca2+ signaling altered by overexpression of Orai3 may be a mechanism whereby oxLDL increases HLEpiC proliferation to contribute to the development of cataract. Thus, Orai3 may be a target warranting development for the treatment of cataract associated with obesity or hyperlipidemia.

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