scholarly journals Quantitative Evaluation of Human Lens Epithelial Cell Viability and Cytolysis by Distilled Water Ex Vivo

2021 ◽  
Author(s):  
Wen-Wen Zhang ◽  
Rong-Pei Zhang ◽  
Ya-Jun Liu ◽  
Zi-Fang He ◽  
Si Zhang ◽  
...  
Keyword(s):  
Cell Viability ◽  
Ex Vivo ◽  
Lens Epithelial Cell ◽  
Human Lens ◽  
Distilled Water ◽  
Histopathological Changes ◽  
Human Lens Epithelial Cell ◽  
Transmission Electron ◽  
The Difference

Abstract Purpose: To investigate the role of distilled water (DW) in isolated human lens epithelial cells (LECs) viability and lysis ex vivo.Methods: After immersion in DW or balanced salt solution (BSS) for 1-, 2-, and 3-minutes, respectively, the cell viability of LECs was quantitatively evaluated. In addition, the capsule samples soaked in DW or BSS for 1-, 2-, and 3-minutes were combined with rinse for 1 minute to analyze the difference of LECs shedding percentage in each subgroup. The histopathological changes of the samples after treating were observed.Results: The percentage of LECs shed in DW immersion combined with rinse was significantly higher than in DW immersion alone (p all <0.001). In the subgroup soaked in DW for 3 minutes, the death number, mortality, and the percentage of cell shedding of LECs was the most (p all <0.001). The histopathological changes showed that the cell destruction in the DW subgroup for 1-, 2-, and 3-minutes, and the transmission electron microscope results showed that the cells were partially detached from the capsule in the DW 3 minutes subgroup.Conclusions: Soaking in the DW can cause LECs death, and DW immersion combined with rinse was an effective method to remove LECs. The histopathology changes of treated DW suggested cellular necrosis was one type of LECs death mechanism.

EAAT and Xc−Exchanger Inhibition Depletes Glutathione in the Transformed Human Lens Epithelial Cell Line SRA 01/04

2015 ◽  
pp. 1-10 ◽  
Author(s):  
Marlyn P. Langford ◽  
Thomas B. Redens ◽  
Chanping Liang ◽  
A. Scott Kavanaugh ◽  
Donald E. Texada
Keyword(s):  
Epithelial Cell ◽  
Cell Line ◽  
Lens Epithelial Cell ◽  
Human Lens ◽  
Epithelial Cell Line ◽  
Human Lens Epithelial Cell

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

2016 ◽  
Vol 2016 ◽  
pp. 1-17 ◽  
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.

scholarly journals Effect of a Lens Protein in Low-Temperature Culture of Novel Immortalized Human Lens Epithelial Cells (iHLEC-NY2)

Cells ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 2670
Author(s):  
Naoki Yamamoto ◽  
Shun Takeda ◽  
Natsuko Hatsusaka ◽  
Noriko Hiramatsu ◽  
Noriaki Nagai ◽  
...  
Keyword(s):  
Amyloid Β ◽  
Lens Epithelial Cell ◽  
Human Lens ◽  
Epithelial Cell Line ◽  
Lens Protein ◽  
Medium Temperature ◽  
Tropical Regions ◽  
Human Lens Epithelial Cell ◽  
In Silico Simulation ◽  
Temperature Ranges

The prevalence of nuclear cataracts was observed to be significantly higher among residents of tropical and subtropical regions compared to those of temperate and subarctic regions. We hypothesized that elevated environmental temperatures may pose a risk of nuclear cataract development. The results of our in silico simulation revealed that in temperate and tropical regions, the human lens temperature ranges from 35.0 °C to 37.5 °C depending on the environmental temperature. The medium temperature changes during the replacement regularly in the cell culture experiment were carefully monitored using a sensor connected to a thermometer and showed a decrease of 1.9 °C, 3.0 °C, 1.7 °C, and 0.1 °C, after 5 min when setting the temperature of the heat plate device at 35.0 °C, 37.5 °C, 40.0 °C, and 42.5 °C, respectively. In the newly created immortalized human lens epithelial cell line clone NY2 (iHLEC-NY2), the amounts of RNA synthesis of αA crystallin, protein expression, and amyloid β (Aβ)1-40 secreted into the medium were increased at the culture temperature of 37.5 °C compared to 35.0 °C. In short-term culture experiments, the secretion of Aβ1-40 observed in cataracts was increased at 37.5 °C compared to 35.0 °C, suggesting that the long-term exposure to a high-temperature environment may increase the risk of cataracts.

scholarly journals Ferulic Acid Suppresses AmyloidβProduction in the Human Lens Epithelial Cell Stimulated with Hydrogen Peroxide

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Noriaki Nagai ◽  
Sachiyo Kotani ◽  
Yu Mano ◽  
Akina Ueno ◽  
Yoshimasa Ito ◽  
...  
Keyword(s):  
Gene Expression ◽  
Ferulic Acid ◽  
Lens Epithelial Cell ◽  
Human Lens ◽  
Lens Epithelial Cells ◽  
Rt Pcr ◽  
Human Lens Epithelial Cell ◽  
Oxidative Stresses ◽  
Age Related ◽  
The Brain

It is well known that oxidative stresses induce the production of amyloidβ(Aβ) in the brain, lens, and retina, leading to age-related diseases. In the present study, we investigated the effects of ferulic acid on the Aβlevels in H2O2-stimulated human lens epithelial (HLE) SRA 01/04 cells. Three types of Aβpeptides (Aβ1-40, Aβ1-42, and Aβ1-43) were measured by ELISA, and the levels of mRNA for the expressed proteins related to Aβproduction (APP, BACE1, and PS proteins) and degradation (ADAM10, NEP, and ECE1 proteins) were determined by quantitative real-time RT-PCR. H2O2stimulation augmented gene expression of the proteins related to Aβproduction, resulting in the production of three types of Aβpeptides. Treatment with 0.1 μM ferulic acid attenuated the augmentations of gene expression and production of the proteins related to the secretion of three types of Aβpeptides in the H2O2-stimulated HLE cells. These results provided evidence of antioxidative functions of ferulic acid for lens epithelial cells.

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