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Objective To detect the expression of miR-138 in lens tissues of agerelated cataract and explore the effects of miR-138 on the proliferation and apoptosis of human lens epithelial cells and its possible target genes.Methods Real-time quantitative PCR (RT-qPCR) was applied for the detection of the expression of miR-138 and prediction of target gene sirtuin (silent information regulator 1) (SIRT1) in patients with age-related cataract (cataract group) and anterior lens capsules (normal control group).Then miR-138 mimics,mimic controls,miR-138 inhibitors and inhibitor controls were transfected into the human lens epithelial cell line (SRA01/04),and the expression of SIRT1 mRNA and protein was detected by RT-qPCR and Western blot,accordingly.At 72 hours after transfection,the cells were exposed to 200 μmol · L-1 H2O2 for 1 hour,followed by detection of the activity of Caspase-3 by the Caspase-3 activity assay kit,and identification of the targeted relationship between miR-138 and SIRT1 by dual luciferase reporter assays.Results Compared with the normal control group,the expression of miR-138(3.64 ±0.19) was significantly increased (P ＜0.001),but the expression of SIRT1 mRNA(0.32 ± 0.06) was significantly decreased (P ＜ 0.001) in the cataract group.Moreover,The expression levels of SIRT1 mRNA(0.42 ± 0.05) and protein(0.46 ± 0.05) in cells transfected with miR-138 mimics were significantly decreased,while the activity of Caspase-3 (3.24 ± 0.17) was significantly elevated when compared with cells transfected with minic controls (all P ＜ 0.05);Compared with cells transfected with inhibitor controls,the expressions of SIRT1 mRNA(2.95 ±0.13) and protein(1.98 ±0.12) were significantly upregulated,whereas Caspase-3 activity(0.42 ±0.05) was significantly decreased in cells transfected with miR-138 inhibitors (all P ＜0.05).And fmally,dual luciferase reporter assays showed the confirmation SIRT1 as a direct target of miR-138.Conclusion miR-138 is highly expressed in the lens capsule of age-related cataract patients,and it can promote the apoptosis of lens epithelial cells by negatively regulating the expression of SIRT1.
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·AIM:To investigate the effects and mechanism of miR-138 in mediating the antioxidant function of lens epithelial cells affected by age-related cataracts. ·METHODS:Real-time quantitative PCR(RT-qPCR) was used to detect miR-138 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 400μ mol/L H2O2for 1h. SRA01/04 cells were transfected with either miR-138 mimics,mimic controls, miR-138 inhibitors or inhibitor controls. After 72h,these cells were exposed to 400μ mol/L H2O2for 1h, then p53 and Bax mRNA expression were measured using RT-qPCR. Expression of p53 and Bax protein 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-138 in the anterior lens capsules of age-related cataract patients and in SRA01/04 cells exposed to oxidative stress 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-138 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-138 inhibitor group as compared to the control group, while the cells in this group had much higher levels of cell viability (P<0.001). · CONCLUSION: The expression of miR - 138 is upregulated in the anterior lens capsules of age-related cataract patients. MiR-138 decreases the anti-oxidative stress capacity of lens epithelial cells by upregulating p53 and Bax, while inhibiting cell proliferation and repair. This finding suggests that miR-138 may play a key role in the development of age-related cataracts.
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SUMOylation is a post - translational modification consisting of covalent conjugation of ubiquitin - like proteins called small ubiquitin related modifier ( SUMO ) . SUMO modification has been shown to significantly alter protein activity, which can modulate protein stability, affect protein-protein interactions, and modify protein localization and trafficking. This process adds another layer of control in eukaryote gene expression, and it regulates both transcriptional activation and repression. This article reviews the current situation and future development of SUMOylation in ophthalmology.
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AlM:To study the relationship between JAK-STAT pathway and epithelial - mesenchymal transition in human lens epithelial cells. Meanwhile, the function of AG490 as a JAK inhibitor was also demonstrated in this article. METHODS:Human lens epithelial cells SRA01/04 ( LECs ) were treated by low concentration of glucose (5. 5mmol/L). High concentration of glucose (30. 5mmol/L) was used to treat the cells in order to form the high glucose model. According to adding AG490 or not, cells were divided into the control group and the experimental group, appropriate concentration 10ü mol/L and 50ü mol/L of AG490 were chosen and acting time of 6, 12, 24, 48h were selected. Effect of AG490 on cell migration was measured by wound healing test. The expression of TGF-β1 , FN,α-SMA mRNA were examined by RT-PCR. RESULTS:With the prolonged acting time ( 6, 12, 24 and 48h), cell activity increased in the HG group, as well as more expression of TGF-β1 , FN,α-SMA mRNA were detected compared to the LG group (P CONCLUSlON:JAK-STAT pathway takes part in high glucose-induced epithelial-mesenchymal transition in human lens epithelial cells. The mechanism is that it impacts the transcriptional expression of TGF- β1 and extracellular matrix. AG490, a JAK inhibitor, inhibits high glucose-induced epithelial-mesenchymal transition in human lens epithelial cells, And the inhibition enhances with the increasing concentration of AG490.
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· AIM: To compare the measurement of anterior chamber depth (ACD) and axial length (AL) by IOLMaster and contact ultrasonic (US) axial scan (A-scan).· METHODS: Measurements of ACD and AL were prospectively obtained in 137 eyes of 121 subjects with the IOLMaster compared with measurements with the US.· RESULTS: There was an excellent correlation between IOLMaster and US measurements for the ACD (r=0.823;P＜0.001) and AL (r=0.996;P＜0.001). The mean values of the parameters measured by IOLMaster and US were,respectively, as follows: ACD, 2.94±0.49mm, 2.58±0.51mm;AL, 24.37±3.04mm, 23.81±2.83mm. The mean differences of ACD and AL values between IOLMaster and US measurements were 0.36 ±0.30mm, 0.56 ±0.34 mm respectively, and they proved to be statistically significant (P＜0.001), With the 95%limits of agreement (LoA) from -0.08mm to +0.38mm for ACD and from -0.09mm to +0.69mm for AL.· CONCLUSION: As noncontact biometry, IOLMaster provides accurate values. A high degree of agreement between US and IOLMaster was noted. It not only has the advantage of performing noncontact examinations, but also produces various additional data simultaneously and may thus obviate the need for multiple examinations. Further studies are needed to assess the interchangeability of measurements in clinical practice.
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AIM: To assess the relative agreement of GAT and NCT in IOP measurement by comparing the differences between Goldmann applanation tonometer (GAT) and non-contact tonometer (NCT) in intraocular pressure (IOP) detection.METHODS: IOP of 529 eyes of 265 volunteers were measured with both NCT and GAT, respectively.RESULTS: The measurement results of NCT were lower than that of GAT, there was significant difference between the IOP measured with NCT and GAT (19.13 vs23.43, t=22.644, P＜0.05). With the increasing of IOP values, the difference magnitude was greater, especially in IOP group that was more than 30mmHg, but the correlation coefficient became lower.CONCLUSION: The measurement results with NCT are lower than that of GAT. When the IOP with the NCT is in borderline value, it need be corrected with GAT, in order to discover the pathologically elevated IOP and avoid the misdiagnosis and mistreatment of glaucoma.