青光眼是一组以视盘萎缩凹陷、视野缺损以及视力下降为共同特征的视神经退行性疾病，也是世界首位不可逆性致盲眼病，导致患者生活质量降低、引起极大卫生经济负担。但其发病机制尚不明确，促进房水排出从而降低眼内压仍是目前减缓疾病进展的唯一治疗手段。房水排出的主要途径是经由小梁网进入Schlemm's管最后汇入巩膜外静脉，因此小梁网在调节房水排出以及平衡眼内压方面发挥重要作用。近年来，体内以及体外房水排出测量技术和小梁网成像技术不断突破，众多研究表明小梁网存在压力依赖的节律性搏动，在房水的脉冲式排出中起到关键作用，但在青光眼中这种搏动随疾病的进展减弱甚至消失。文章以小梁网的泵理论为核心，总结青光眼中房水排出的最新研究进展，并从恢复小梁网功能的角度出发探索可能有效的治疗策略，为青光眼的临床诊治提供新的思路。
Glaucoma, a group of optic nerve degenerative diseases, is characterized by papillary atrophy, visual field defects, and decreased vision. It is also the leading cause of irreversible blindness worldwide, significantly reducing patients’ the quality of life of patients and posing considerable health economic burdens. However, the pathogenesis of glaucoma remains unclear, and promoting aqueous humor outflow to reduce intraocular pressure is the only treatment option available to slow disease progression. The main pathway for aqueous humor outflow is through the trabecular meshwork into Schlemm's canal and finally into the episcleral veins, highlighting the crucial role of the trabecular meshwork in regulating aqueous humor outflow and maintaining intraocular pressure balance. In recent years, there have been notable breakthroughs in in vivo and in vitro aqueous humor outflow measurement techniques and trabecular meshwork imaging technologies.Many studies suggest that the trabecular meshwork exhibits pressure-dependent rhythmic pulsation, playing a crucial role in the pulse-like outflow of aqueous humor. Unfortunately, in glaucoma, this pulsation weakens or even disappears as the disease progresses. This article focuses on the trabecular meshwork's pump theory and summarizes the latest research progress in aqueous humor outflow in glaucoma, exploring potential effective therapeutic strategies aimed at restoring trabecular meshwork function. This provides new insights for the clinical diagnosis and treatment of glaucoma.
目的：通过在人小梁网细胞(human trabecular meshwork cell，HTMC)中过表达沉默信息调节因子2相关酶1(silent information regulator 1，SIRT1)，探讨SIRT1对氧化应激下HTMC功能的影响。方法：将SIRT1过表达慢病毒和GFP阴性对照慢病毒按照最佳(multiplicity of infection，MOI)分别转染入HTMC，并用实时定量PCR法对SIRT1是否在细胞中过表达进行验证。实验分为以下4组：正常组、H2O2组、H2O2+Lv-SIRT1-OE(过表达)组、H2O2+Lv-GFP组，分别采用Transwell法和CCK8法检测氧化应激下HTMC的迁移能力和活性。两组间比较采用独立样本t检验。结果：在正常组、H2O2组、H2O2+Lv-SIRT1-OE组、H2O2+Lv-GFP组这4组中，Transwel l实验结果分别为436±73、254±25、510±51、327±46，H2O2+Lv-SIRT1-OE组分别与H2O2组和H2O2+Lv-GFP组差异均有统计学意义(P<0.01)。CCK8法结果显示，H2O2+Lv-SIRT1-OE组分别与H2O2组和H2O2+Lv-GFP组相比差异均有统计学意义(P<0.01)。H2O2+Lv-SIRT1-OE组分别与H2O2组和阴性对照组(H2O2+Lv-GFP)相比，Bax表达水平明显下降，Bcl-2表达水平明显提高，差异均有统计学意义(P<0.01)。ROS活性氧测定显示H2O2+Lv-SIRT1-OE组比H2O2组的细胞活性氧水平显著降低(P<0.05)。结论：在HTMC中过表达SIRT1能有效降低氧化应激对HTMC迁移能力和活性的影响，从而对HTMC起到一定的保护作用，为后续研究SIRT1保护氧化应激下HTMC的调控机制打下基础。
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 signifificance. 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.
