Effect of SIRT1 on cell function of human trabecular meshwork cell under oxidative stress
Journal Title: Eye Science (Yanke Xuebao) - Year 2021, Vol 36, Issue 6
Abstract
[Objective:] To explore the effect of Silent Information Regulator 1 (SIRT1) on cell function of human trabecular meshwork cell (HTMC) under oxidative stress by overexpressing SIRT1 in HTMC. [Methods:] This is an experiment research. HTMCs were transfected with SIRT1-ovexpressed lentivirus and GFP-negative control lentivirus (Lv-GFP) at the optimal multiplicity of infection (MOI). Real-time quantitative PCR was used to verify whether SIRT1 was overexpressed in HTMC. The following experiments were divided into four groups: normal control group, H2O2 group, H2O2+Lv-SIRT1-OE group, H2O2+Lv-GFP group. Cell migration was detected by transwell assay. Cell viability was detected by CCK8 assay. Student's t-test was used for two groups. P<0.05 was set as statistical significance. [Results:] The number of migration per well of normal control group, H2O2 group, H2O2+Lv-SIRT1-OE group, H2O2+Lv-GFP group were 436±73, 254±25, 510±51, 327±46, respectively. Compared with H2O2 group and H2O2+Lv-GFP group, transwell assay demonstrated that the number of migrations per well of H2O2+Lv-SIRT1-OE group significantly increased (P<0.01). Likewise, CCK8 assay indicated that cell viability of H2O2+Lv-SIRT1-OE group was higher than both of H2O2 group and H2O2+Lv-GFP group (P<0.01). Compared with H2O2+Lv-SIRT1-OE group and negative control group (H2O2+Lv-GFP), the expression level of Bax decreased significantly, and the expression level of Bcl-2 increased significantly (P<0.01). ROS assay showed that the ROS level in H2O2+Lv-SIRT1-OE group was significantly lower than that in H2O2 group (P<0.05). [Conclusion:] SIRT1 overexpressed in HTMC can effectively reduce the effect of oxidative stress on migration ability and proliferation activity of HTMC, which lays a foundation for further study on the regulatory mechanism of SIRT1 protecting HTMC under oxidative stress.
Authors and Affiliations
Junhong GUO, Xiaoli SHEN, Xiangqian HONG, Dahui MA, Hongbin XIE, Mingmin YANG, Jiantao WANG
Keywords
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- EP ID EP763519
- DOI 10.3978/j.issn.1000-4432.2021.06.16
- Views 15
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