转化生长因子-β1诱导培养的人眼小梁细胞产生弹性蛋白的实验研究

目的 了解转化生长因子－β１（ｔｒａｎｓｆｏｒｍｉｎｇ ｇｒｏｗｔｈ ｆａｃｔｏｒ－β１,ＴＧＦ－β１）诱导培养培养的人眼小梁细胞产生的弹性蛋白。探讨房水引流阻力在原发性开角型青光眼（ｐｒｉｍａｒｙ ｏｐｅｎ ａｎｇｌｅ ｇｌａｕｃｏｍａ,ＰＯＡＧ）的病因和发病机制中的应用。方法 小梁网来源于除眼部疾患外的各种原因死亡的６岁以下儿童,在２４ｈ内取才于６例（１２只眼）正常眼球。人眼小梁细胞原代和传代

Transforming growth factor-beta1 induced cultured human trabecular cells to produce elastin

OBJECTIVE: To understand transforming growth factor-beta(1) (TGF-beta(1)) inducing cultured human trabecular cells (HTCs) to produce elastin and to approach the effect of resistance of aqueous outflow in the pathogenesis and mechanism of primary open angle glaucoma (POAG). METHODS: Trabecular meshwork's specimens were collected from twelve normal eyeballs of six human donors under six years of age within 24 hours after death. HTCs were primarily cultured and subcultured. Cultured cells were observed under light and electron microscopes. Laminin (LN) and IV collagen (IV C) in extracellular matrix (ECM) were immunohistochemically stained by S-P method. The third passage confluent cells after two weeks were divided into three groups: control group, TGF-beta(1) group and neutralizing antibody of TGF-beta(1) group. Concentration of neutralizing antibody of TGF-beta(1) was 30 microg/ml. After culturing for two weeks, elastin in the media of three groups was respectively determined by western blotting from sodium dodecylsulfate-polyacrylamide gel electrophoresis using anti-human aortic alpha-elastin rabbit serum, biotin conjugated goat anti-rabbit IgG and anti-biotin conjugated horse radish peroxidase. RESULTS: Cultured HTCs were identified from cultured cell's growing characteristics, morphological features under light and electron microscopes, and stained peculiarities of LN and IV C in ECM. With western blotting, elastin in the control group was not detected, elastin in TGF-beta(1) group was detected, and elastin in neutralizing antibody of TGF-beta(1) group was not detected. CONCLUSION: TGF-beta(1) induces cultured HTCs to produce elastin, which the neutralizing antibody of elastin can antagonize; the increase of the amount of elastin may aggravate its stacking in ECM and increase the resistance of aqueous outflow in trabecular meshwork, leading to the occurrence of glaucoma.