Dexamethasone inhibition of trabecular meshwork cell phagocytosis and its modulation by glucocorticoZhang X, Ognibene CM, Clark AF, Yorio T. Department of Pharmacology and Neuroscience, University of North Texas Health Science Center, 3500 Camp Bowie Boulevard, Fort Worth, TX 76107, USA.Glucocorticoid treatment can lead to the development of glaucomatous ocular hypertension and a secondary open-angle glaucoma due to increased aqueous humor outflow resistance that is associated with morphological and biochemical changes in the trabecular meshwork (TM). The cellular responses of glucocorticoids are achieved by binding to the glucocorticoid receptor alpha (GRalpha), a ligand-activated transcription factor. An alternatively spliced variant, glucocorticoid receptor beta (GRbeta), has dominant negative activity on GRalpha and has been implicated in a variety of steroid-resistant diseases. We previously showed that GRbeta can block dexamethasone (DEX) responsiveness in TM cells. TM cells are actively phagocytic and function in the removal of debris, pigment and other materials from the aqueous outflow drainage pathway. A decrease in phagocytic activity has been proposed in the pathogenesis of glaucoma and glucocorticoid-induced glaucoma. In this study, we investigated the effect of DEX and GRbeta on phagocytosis in normal and glaucomatous TM cells. Human transformed normal NTM-5 and primary normal NTM174-00 cells, which express relatively high amounts of GRbeta, and transformed glaucomatous GTM-3 and primary glaucomatous GTM520-05 cells, which have lower GRbeta expression, were treated with 100 nM DEX or vehicle control for 24h. NTM cells also were transfected with a control or GRbeta expression plasmid to examine the effect of GRbeta on phagocytic activity. The cells were incubated with Alexa 488 conjugated Staphylococcus aureus bioparticles opsonized with rabbit IgG for 1h, followed by fixation and incubation with Alexa 633 conjugated goat anti-rabbit IgG to distinguish ingested from extracellular bioparticles. DAPI nuclear staining was used to quantify cell numbers. Cells and bioparticles were visualized by confocal microscopy. We found that NTM-5 cells ingested more bioparticles than GTM-3 cells. DEX treatment significantly decreased the phagocytosis of bioparticles in NTM-5 and GTM-3 cells, while GTM-3 cells were more responsive to DEX, compared to NTM-5 cells. In primary cell culture, NTM174-00 also engulfed more bioparticles than GTM520-05 cells. DEX treatment significantly decreased the phagocytic activity in GTM520-05, but not in NTM174-00 cells. Transient transfection of pCMX-hGRbeta plasmid increased the expression of GRbeta and consequently maintained the phagocytotic activity of NTM-5 cells in the presence of DEX. Our data demonstrated that the expression level of GRbeta in TM cells can regulate DEX-induced suppression of phagocytotic activity. The lower expression of GRbeta in glaucomatous TM cells may contribute to the altered phagocytic function of TM cells, and may lead to the increased aqueous humor outflow resistance mediated by glucocorticoids.PMID: 17126833 [PubMed - indexed for MEDLINE]本人已认领该文编译，48小时后若未提交译文，请其他战友自由认领Exp Eye Res. 2007 Feb;84(2):275-84. Epub 2006 Nov 28.题目：Dexamethasone inhibition of trabecular meshwork cell phagocytosis and its modulation by glucocorticoid receptor beta.地塞米松对小梁网细胞吞噬功能的抑制作用和糖皮质激素受体-β对它的调节作者：Zhang X, Ognibene CM, Clark AF, Yorio T. 作者单位：Department of Pharmacology and Neuroscience, University of North Texas Health Science Center, 3500 Camp Bowie Boulevard, Fort Worth, TX 76107, USA.文摘：Glucocorticoid treatment can lead to the development of glaucomatous ocular hypertension and a secondary open-angle glaucoma due to increased aqueous humor outflow resistance that is associated with morphological and biochemical changes in the trabecular meshwork (TM). 激素可引起小梁网在形态学和生化学上发生改变，使房水流出阻力增加，从而导致激素性高眼压和继发性开角性青光眼。The cellular responses of glucocorticoids are achieved by binding to the glucocorticoid receptor alpha (GRalpha), a ligand-activated transcription factor. 糖皮质激素通过与配体依赖转录因子糖皮质激素受体-α（GRα）结合来发挥作用。An alternatively spliced variant, glucocorticoid receptor beta (GRbeta), has dominant negative activity on GRalpha and has been implicated in a variety of steroid-resistant diseases.GRβ由GR基因的同一转录产物通过另一种剪切方式产生，在许多激素抵抗性疾病的研究中显示它对GRα的功能有拮抗作用。We previously showed that GRbeta can block dexamethasone (DEX) responsiveness in TM cells.在前期研究中我们已经发现GRβ可以阻断地塞米松对小梁网细胞的作用。TM cells are actively phagocytic and function in the removal of debris, pigment and other materials from the aqueous outflow drainage pathway.小梁网细胞具有吞噬功能，能够清除房水流出途径中所产生的细胞碎片、色素和其他物质。A decrease in phagocytic activity has been proposed in the pathogenesis of glaucoma and glucocorticoid-induced glaucoma. 在青光眼和激素性青光眼的发病机制的研究中已经发现小梁网细胞的吞噬功能有所下降。In this study, we investigated the effect of DEX and GRbeta on phagocytosis in normal and glaucomatous TM cells. 本实验我们将研究地塞米松和GRβ对正常人群和青光眼人群的小梁网细胞吞噬功能的影响。Human transformed normal NTM-5 and primary normal NTM174-00 cells, which express relatively high amounts of GRbeta, and transformed glaucomatous GTM-3 and primary glaucomatous GTM520-05 cells, which have lower GRbeta expression, were treated with 100 nM DEX or vehicle control for 24h. 人们将正常的NTM-5细胞和原代培养的正常NTM174-00细胞进行转化，使其高表达GRβ。并用100 nM的地塞米松或媒介物处理青光眼患者的GTM-3细胞和原代GTM520-05细胞，使其表达GRβ的水平降低。NTM cells also were transfected with a control or GRbeta expression plasmid to examine the effect of GRbeta on phagocytic activity. 也可以用操纵子或表达 GRβ的质粒转染NTM细胞的方法来检测GRβ对小梁网细胞吞噬功能的影响。The cells were incubated with Alexa 488 conjugated Staphylococcus aureus bioparticles opsonized with rabbit IgG for 1h, followed by fixation and incubation with Alexa 633 conjugated goat anti-rabbit IgG to distinguish ingested from extracellular bioparticles. 将兔抗IgG 作用于Alexa 488金黄色葡萄球菌的活性颗粒，后与NTM细胞孵育1小时，再用Alexa 633羊抗兔IgG定位，以区别所吞噬的位于细胞外的活性颗粒。DAPI nuclear staining was used to quantify cell numbers. Cells and bioparticles were visualized by confocal microscopy. 用DAPI对细胞核进行染色以计数细胞，用共焦显微镜观察细胞和活性颗粒。We found that NTM-5 cells ingested more bioparticles than GTM-3 cells. DEX treatment significantly decreased the phagocytosis of bioparticles in NTM-5 and GTM-3 cells, while GTM-3 cells were more responsive to DEX, compared to NTM-5 cells. 实验结果显示，NTM-5细胞较GTM-3细胞吞噬更多的活性颗粒。用地塞米松处理后，NTM-5细胞和GTM-3细胞所吞噬的活性颗粒明显减少，并且GTM-3细胞更为敏感。In primary cell culture, NTM174-00 also engulfed more bioparticles than GTM520-05 cells. DEX treatment significantly decreased the phagocytic activity in GTM520-05, but not in NTM174-00 cells. 在原代培养中，NTM174-00 细胞较GTM520-05细胞也吞噬了更多的活性颗粒，地塞米松显著降低了GTM520-05细胞的吞噬功能，但对NTM174-00 细胞没有影响。Transient transfection of pCMX-hGRbeta plasmid increased the expression of GRbeta and consequently maintained the phagocytotic activity of NTM-5 cells in the presence of DEX.pCMX-hGRbeta质粒转染小梁网细胞可以增加GRβ的表达，从而维持在地塞米松作用下NTM-5细胞的吞噬功能。Our data demonstrated that the expression level of GRbeta in TM cells can regulate DEX-induced suppression of phagocytotic activity. 我们的实验数据证明，小梁网细胞的GRβ表达水平可以对地塞米松诱导的吞噬功能的抑制进行调节。The lower expression of GRbeta in glaucomatous TM cells may contribute to the altered phagocytic function of TM cells, and may lead to the increased aqueous humor outflow resistance mediated by glucocorticoids.激素性青光眼患者的小梁网细胞低表达GRβ可能引起小梁网细胞吞噬功能发生改变，从而产生激素介导的房水流出阻力增加。peggy83422 翻译的很好，几点建议如下：开角性青光眼 应为 开角型青光眼vehicle control 是指溶解地塞米松的溶质，这里可以翻作 空白对照 兔抗IgG 可以加一个字，按照上下文意思，应该是兔抗人IgG抗体继续努力！Human transformed normal NTM-5 and primary normal NTM174-00 cells, which express relatively high amounts of GRbeta, and transformed glaucomatous GTM-3 and primary glaucomatous GTM520-05 cells, which have lower GRbeta expression, were treated with 100 nM DEX or vehicle control for 24h. 人们将正常的NTM-5细胞和原代培养的正常NTM174-00细胞进行转化，使其高表达GRβ。并用100 nM的地塞米松或媒介物处理青光眼患者的GTM-3细胞和原代GTM520-05细胞，使其表达GRβ的水平降低。原文可改为：Human transformed normal NTM-5 and primary normal NTM174-00 cells and transformed glaucomatous GTM-3 and primary glaucomatous GTM520-05 cells were treated with 100 nM DEX or vehicle control for 24h. The cells were incubated with Alexa 488 conjugated Staphylococcus aureus bioparticles opsonized with rabbit IgG for 1h, followed by fixation and incubation with Alexa 633 conjugated goat anti-rabbit IgG to distinguish ingested from extracellular bioparticles. 将兔抗IgG 作用于Alexa 488金黄色葡萄球菌的活性颗粒，后与NTM细胞孵育1小时，再用Alexa 633羊抗兔IgG定位，以区别所吞噬的位于细胞外的活性颗粒。to distinguish ingested (bioparticles) from extracellular bioparticles.感谢yooki战友的指点！ 请问您的意思是不是这样的：Human transformed normal NTM-5 and primary normal NTM174-00 cells, which express relatively high amounts of GRbeta, and transformed glaucomatous GTM-3 and primary glaucomatous GTM520-05 cells, which have lower GRbeta expression, were treated with 100 nM DEX or vehicle control for 24h. 高表达GRβ的转化正常人NTM-5细胞和原代正常人NTM174-00细胞，以及低表达GRβ的转化青光眼患者GTM-3细胞和青光眼患者GTM520-05细胞，均被用100nM的地塞米松或空白对照载体处理24h。The cells were incubated with Alexa 488 conjugated Staphylococcus aureus bioparticles opsonized with rabbit IgG for 1h, followed by fixation and incubation with Alexa 633 conjugated goat anti-rabbit IgG to distinguish ingested from extracellular bioparticles. 将兔抗IgG 作用于Alexa 488金黄色葡萄球菌的活性颗粒，后与实验细胞(上述4种)孵育1小时，再用Alexa 633羊抗兔IgG定位，以区别活性颗粒是被吞噬的还是位于细胞外的。学习学习！非常感谢各位！