UNASSIGNED:为了确定人类小梁网(TM)中氧化损伤和应激以及线粒体功能的种族差异。
未经评估:实验研究。
未经评估:173名接受眼内手术的患者的一百七十三只眼睛提供了房水(AH)进行分析。来自眼库供体的小梁网组织用作原代细胞培养的健康对照。
UNASSIGNED:酶联免疫吸附测定法用于测量8-羟基-2-脱氧鸟苷(8-OHdG),氧化损伤标记,在AH比较美国黑人和白人。在高氧和低氧培养条件下,将来自黑白供体的人TM原代培养细胞用于三磷酸腺苷(ATP)测量。在从培养的TM细胞分离的线粒体级分中测量复合物I活性。通过线粒体外膜转位酶20(TOMM20)Western印迹进行线粒体定量。在活TM细胞中测量细胞内活性氧(ROS)产生。
未经评估:AH中的氧化损伤,ATP生产,复杂的I活动,线粒体定量,和按种族背景分层的培养TM细胞中的细胞内ROS。
未经批准:房水样本(75黑色,98White)与患有严重青光眼的White患者相比,Black的8-OHdG水平显着升高(P=0.024)。使用培养的健康供体TM细胞,在低氧培养条件下,黑色TM细胞的ATP产量高于白色TM细胞(P=0.002)。与白色TM细胞相比,黑色的复合物I活性没有统计学差异,但TOMM20在黑白细胞中的表达较高(P=0.001)。为了应对过氧化氢的挑战,与白色TM细胞相比,黑色细胞中的ROS产生显著更高(P=0.004)。
UASSIGNED:与患有严重青光眼的白人患者相比,黑人AH患者的8-OHdG水平明显升高,这表明氧化损伤可能是青光眼发病机制的危险因素或黑人人群中不同病理特征的结果。为了确定这种损害的潜在来源或原因,我们的数据显示,健康的黑培养的TM细胞具有较高的ATP和ROS水平,随着线粒体数量的增加,与白色TM细胞相比。这些发现表明线粒体改变和氧化应激增加可能影响青光眼的种族差异。
UNASSIGNED: To identify racial differences of oxidative damage and stress and mitochondrial function in human trabecular meshwork (TM).
UNASSIGNED: Experimental study.
UNASSIGNED: One hundred seventy-three eyes of 173 patients undergoing intraocular surgery provided aqueous humor (AH) for analysis. Trabecular meshwork tissues from eye bank donors were used as healthy controls for primary cell culture.
UNASSIGNED: Enzyme-linked immunosorbent assay methods were used to measure 8-hydroxy-2-deoxyguanosine (8-OHdG), an oxidative damage marker, in AH comparing Black and White Americans. Human TM primary cultured cells from Black and White donors were used for adenosine triphosphate (ATP) measurement under high and low oxygen culture conditions. Complex I activity was measured in mitochondrial fractions isolated from cultured TM cells. Mitochondrial quantification was performed by translocase of outer mitochondrial membrane 20 (TOMM20) Western blot. Intracellular reactive oxygen species (ROS) production was measured in live TM cells.
UNASSIGNED: Oxidative damage in AH, ATP production, complex I activity, mitochondrial quantification, and intracellular ROS in cultured TM cells stratified by racial background.
UNASSIGNED: Aqueous humor samples (75 Black, 98 White) displayed significantly higher 8-OHdG levels (P = 0.024) in Black compared with White patients with severe stage glaucoma. Using cultured healthy donor TM cells, ATP production was higher in Black than White TM cells (P = 0.002) in low oxygen culture conditions. Complex I activity was not statistically different in Black compared with White TM cells, but TOMM20 expression was higher in Black versus White cells (P = 0.001). In response to hydrogen peroxide challenge, ROS production was significantly higher in Black compared to White TM cells (P = 0.004).
UNASSIGNED: Significantly higher 8-OHdG levels in AH of Black compared with White patients with severe glaucoma indicated that oxidative damage may be a risk factor in glaucoma pathogenesis or the result of distinct pathologic features in the Black population. To identify potential origins or causes of this damage, our data showed that healthy Black cultured TM cells have higher ATP and ROS levels, with increased quantity of mitochondria, compared with White TM cells. These findings indicate that mitochondrial alterations and increased oxidative stress may influence racial disparities of glaucoma.