miR-211 regulates the antioxidant function of lens epithelial cells affected by age-related cataracts

AIM： To investigate the effects and mechanism of miR-211 in mediating the antioxidant function of lens epithelial cells affected by age-related cataracts. METHODS： Real-time quantitative polymerase chain reaction （RT-qPCR） was used to detect miR-211 expression in the anterior lens capsules of healthy people, the anterior lens capsules of patients with age-related cataracts, and human epithelial cell line （SRA01/04） cells exposed to oxidative stress. A 2＇, 7＇-dichloro-fluorescein diacetate （DCFH-DA） probe was used to measure the levels of endogenous reactive oxygen species （ROS） in human lens epithelial cells （hLECs） exposed to 400 pmol/L H2O2 for lh. SRA01/04 cells were transfected with either miR-211 mimics, mimic controls, miR-211 inhibitors or inhibitor controls. After 72h, these cells were exposed to 400 IJmollL H2O2 for lh, then p53 and Bax mRNA expression were measured using RT-qPCR. p53 and Bax protein expression were also measured by Western blotting analysis. Finally, cell viability was assessed using an MTS assay. RESULTS： Compared to the control group, expression of miR-211 in the anterior lens capsules of age-related cataract patients and in SRA01/04 cells exposed to oxidative stress was significantly increased （P〈0.001）. Levels of endogenous ROS were significantly elevated in hLECs exposed to oxidative stress （P〈0.001）. Compared to the mimic control group, the hLECs in the miR-211 mimic group expressed significantly higher levels of p53 and Bax mRNA and protein while cell viability was significantly reduced （P〈0.001）. Conversely, p53 and Bax mRNA and protein expression were significantly reduced in the miR-211 inhibitor group as compared to the control group, while the cells in this group had much higher levels of call viability （P〈0.001）. CONCLUSION： miR-211 is upregulatsd in the anterior lens capsules of age-related cataract patients, miR-211 decreased the antioxidative stress capacity of lens epithelial cells by upregulating p53 and Bax, while inhibiting cell proliferation and repair. This finding suggests that miR-211 may play a key role in the development of age-related cataracts.

SUMOylation and deacetylation affect NF-κB p65 activity induced by high glucose in human lens epithelial cells

AIM: To explore the effects of IκBα SUMOylation and NF-κB p65 deacetylation on NF-κB p65 activity induced by high glucose in cultured human lens epithelial cells(HLECs).METHODS: HLECs(SRA01/04) were cultured with 5.5, 25, and 50 mmol/L glucose media for 24 h, and with 50 mmol/L glucose media for 0, 12, and 24 h respectively. SUMO1 and SIRT1 expressions were detected by reverse transcriptionpolymerase chain reaction(RT-PCR) and Western blot(WB). IκBα and NF-κB p65 expressions were detected by WB. With NAC, DTT, MG132 or Resveratrol(RSV) treatment, SUMO1 and SIRT1 expressions were detected by WB. Protein expression localizations were examined by immunofluorescence and co-immunofluorescence. The effects of SUMO1 or SIRT1 overexpression, as well as MG132 and RSV, on the nuclear expression and activity of IκBα and NF-κB p65 were analyzed by immunoblot and dual luciferase reporter gene assay.RESULTS: SUMO1 and SIRT1 expressions were influenced by high glucose in mRNA and protein levels, which could be blocked by NAC or DTT. SUMO1 was down-regulated by using MG132, and SIRT1 was up-regulated under RSV treatment. IκBα nuclear expression was attenuated and NF-κB p65 was opposite under high glucose, while IκBα and NF-κB p65 location was transferred to the nucleus. SUMO1 or SIRT1 overexpression and MG132 or RSV treatment affected the nuclear expression and activity of IκBα and NF-κB p65 under high glucose condition.CONCLUSION: IκBα SUMOylation and NF-κB p65 deacetylation affect NF-κB p65 activity in cultured HLECs under high glucose, and presumably play a significant role in controlling diabetic cataract.

Prevalence and risk factors of lens opacities in rural populations living at two different altitudes in China

AIM： To investigate the prevalence of and risk factors for lens opacities in populations living at two different altitudes in China.·METHODS： A total of 813 subjects aged ≥40y in Lhasa（Tibet Autonomous Region, China. Altitude： 3658 m） and Shaoxing（Zhejiang Province, China. Altitude： 15 m） were underwent eye examinations and interviewed in this cross-sectional study. Participants＇ lens opacities were graded according to the Lens Opacities Classification System II（LOCS II） and the types of opacities with LOCS II scores ≥2 were determined. Univariate and stepwise logistic regression were used to evaluate the associations of independent risk factors with lens opacities.· RESULTS： Lens opacities were significantly more prevalent in the high-altitude than in the low-altitude area（χ2=10.54, P 〈0.001）. Lens opacities appear to develop earlier in people living at high than at low altitude. The main types of lens opacity in Lhasa and Shaoxing were mixed（23.81%） and cortical（17.87%）,respectively. Independent risk factors associated with all lens opacities were age, ultraviolet（UV） radiation exposure,and educational level. Compared with participants aged40-49 y, the risk of lens opacities increased gradually from 2 to 85 times per 10 y [odds ratio（OR）=2.168-84.731,P 〈0.05）. The risk of lens opacities was about two times greater in participants with the highest UV exposure than in those with the lowest exposure（OR=2.606, P =0.001）.Educational level was inversely associated with lensopacities; literacy deceased the risk by about 25%compared with illiteracy（OR=0.758, P =0.041）.·CONCLUSION： Old age, higher UV exposure and lower educational level are important risk factors for the development of lens opacities. Lens opacities are more prevalent among high-altitude than low-altitude inhabitants.

miR-211 promotes lens epithelial cells apoptosis by targeting silent mating-type information regulation 2 homolog 1 in age-related cataracts

AIM： To detect the expression of miR-211 in age-related cataract tissue, explore the effects of miR-211 on lens epithelial cell proliferation and apoptosis, and identify its target gene.METHODS： This study used real-time quantitative polymerase chain reaction（RT-q PCR） to measure the expression of miR-211 and its predicted target gene [silent matingtype information regulation 2 homolog 1（SIRT1）] in 46 anterior lens capsules collected from age-related cataract patients. Human lens epithelial cell line（SRA01/04） cells were transfected with either miR-211 mimics, mimic controls, miR-211 inhibitors or inhibitor controls, 72 h after transfection, miR NA and protein expression of SIRT1 were measured using RT-qP CR and Western blotting; then cells were exposed to 200 μmol/L H2O2 for 1h, whereupon cell viability was measured by MTS assay, caspase-3 assay was performed. Dual luciferase reporter assay was performed to verify the relationship between miR-211 of SIRT1.RESULTS： Compared to the control group, expression of miR-211 was significantly increased（P〈0.001）, the miR NA and protein expression of SIRT1 were significantly decreased（P〈0.001） in the anterior lens capsules of patients with age-related cataracts. Relative to the control group, SIRT1 miR NA and protein levels in the miR-211 mimic group were significantly reduced, cell proliferation activity significantly decreased, and caspase-3 activity was significantly increased（P〈0.001）. In the miR-211 inhibitor group, SIRT1 miRNA and protein expression were significantly increased, cell proliferation activity significantly increased, and caspase-3 activity was significantly decreased（P〈0.001）. A dual luciferase reporter assay confirmed that SIRT1 is a direct target of miR-211.CONCLUSION： miR-211 is highly expressed in the anterior lens capsules of patients with age-related cataracts. By negatively regulating the expression of SIRT1, miR-211 promotes lens epithelial cell apoptosis and inhibits lens epithelial cell proliferation.

Comparison of 25 MHz and 50 MHz ultrasound biomicroscopy for imaging of the lens and its related diseases

AIM： To compare the results of 25 MHz and 50 MHz ultrasound biomicroscopy（UBM） regarding the image characteristics of the lens and its related diseases and to discuss the application value of 25 MHz UBM in ophthalmology. METHODS： A total of 302 patients（455 eyes） were included in this study from November 2014 to May 2015. Patient ages ranged from 5 to 89 y（mean±SD： 61.0±17.7 y）. Different cross-sectional images of the lens were collected to compare and analyze the image characteristics and anterior segment parameters using 25 MHz and 50 MHz UBM in axial and longitudinal scanning modes, respectively. SPSS 19.0 for Windows, paired t-tests and B＆A plot analysis were used for data analysis, and a value of P〈0.05 was considered statistically significant. RESULTS： The 25 MHz UBM images displayed the lens shape more clearly than 50 MHz UBM images. Particularly for cataracts, the whole opacity of the lens was shown by 25 MHz UBM, but 50 MHz UBM only showed part of the lens. The means of the anterior segment parameters obtained using 25 MHz and 50 MHz UBM were as follows： central corneal thickness： 0.55±0.03 and 0.51±0.04 mm, respectively; central anterior chamber depth： 2.48±0.54 and 2.56±0.56 mm, respectively; and central lens thickness： 4.26±0.62 and 4.15±0.56 mm, respectively. A statistically significant difference was found between the results obtained with 25 MHz UBM and those obtained with 50 MHz UBM. The two devices had a good agreement in measuring the anterior segment parameters. CONCLUSION： The 25 MHz UBM had an obvious advantage in showing the lens shape. It can provide reliable imaging of the lens and its related diseases and has a high application value for ophthalmology.