Matsumoto Y, Johnson D H
Department of Ophthalmology, Mayo Clinic, Rochester, MN 55905, USA.
Invest Ophthalmol Vis Sci. 1997 Aug;38(9):1902-7.
To determine the effect of dexamethasone on the phagocytic capacity of the human trabecular meshwork. A decrease in meshwork phagocytosis has been proposed in the pathogenesis of steroid glaucoma.
The anterior segments of seven pairs of human eyes were placed in perfusion organ culture. One eye of each pair received dexamethasone, and the fellow eye served as control. After 21 days, latex microspheres labeled with fluorescein isothiocyanate and coated with antibodies were added to the culture medium. Twenty-four hours later, the eyes were fixed, the trabecular meshworks were treated with a rhodamine-labeled secondary antibody and sectioned, and the number of ingested beads was determined using a laser scanning confocal microscope. Nuclei were counted and used to calculate the phagocytic index of each anterior segment (number of ingested beads divided by number of nuclei).
Ingested beads appeared green and could be differentiated from noningested beads, which appeared red, using appropriate wavelengths of the laser. Bead ingestion was confirmed with electron microscopy and the use of secondary antibody labeled with horseradish peroxidase. Dexamethasone decreased phagocytosis by 57%, as shown by the fact that trabecular cells in dexamethasone-treated meshworks ingested significantly fewer beads than cells in fellow control meshworks (1.5 +/- 0.6 beads/cell versus 3.5 +/- 1.4 beads/cell; P = 0.008). No evidence of significant migration or loss of trabecular cells was noted; the number of trabecular cells appeared similar in dexamethasone-treated and control meshworks (144 +/- 36 versus 141 +/- 46).
Dexamethasone inhibits phagocytosis by human trabecular meshwork cells in perfusion organ culture.
确定地塞米松对人小梁网吞噬能力的影响。在类固醇性青光眼的发病机制中,有人提出小梁网吞噬作用降低。
将七对人眼的前段置于灌注器官培养中。每对眼睛中的一只接受地塞米松,另一只作为对照。21天后,将用异硫氰酸荧光素标记并包被抗体的乳胶微球加入培养基中。24小时后,将眼睛固定,小梁网用罗丹明标记的二抗处理并切片,使用激光扫描共聚焦显微镜确定摄取珠子的数量。对细胞核进行计数,并用于计算每个前段的吞噬指数(摄取珠子的数量除以细胞核的数量)。
摄取的珠子呈绿色,使用激光的适当波长可与未摄取的珠子区分开来,未摄取的珠子呈红色。通过电子显微镜和使用辣根过氧化物酶标记的二抗证实了珠子的摄取。地塞米松使吞噬作用降低了57%,这表现为地塞米松处理的小梁网中的小梁细胞摄取珠子的数量明显少于对照小梁网中的细胞(1.5±0.6个珠子/细胞对3.5±1.4个珠子/细胞;P = 0.008)。未发现小梁细胞有明显迁移或丢失的证据;地塞米松处理的小梁网和对照小梁网中的小梁细胞数量相似(144±36对141±46)。
在灌注器官培养中,地塞米松抑制人小梁网细胞的吞噬作用。