摘要
背景特发性眼眶炎性假瘤（IOIP）是一种常见的眼眶疾病，病因和发病机制尚未完全阐明，以糖皮质激素为主的治疗效果欠佳。目的观察地塞米松对IOIP组织体外培养成纤维细胞的作用，探讨糖皮质激素在IOIP治疗中可能的作用机制。方法收集2011年11月至2012年1月在北京同仁医院眼科中心切取的6例6眼IOIP组织（IOIP组）和3例因泪腺脱垂行泪腺复位术患者的眶部结缔组织（对照组），采用组织块培养法对2种组织进行体外培养以获取眼眶成纤维细胞，对培养的细胞进行组织形态学观察，采用免疫组织化学法检测培养细胞表面生物标志物的表达；采用WST-8法观察2个组培养的成纤维细胞的增生情况，并绘制生长曲线。将培养的细胞接种至96孔板，将不同浓度地塞米松（O、1×10-3、1×10-4、1×10-5和1×10-6mol／L）加入培养板分别作用24、48和72h，采用WST-8法观察地塞米松对细胞增生的影响；采用ELISA法检测各组成纤维细胞中ICAM-1的相对表达量。结果IOIP组织和正常眼眶体外培养的成纤维细胞均呈长梭形，两端有较长突起；免疫细胞化学染色可见2种组织中培养的细胞中vimentin呈阳性表达，而desmin、S-100、细胞角蛋白（CK）均呈阴性表达。与正常眼眶组织培养的成纤维细胞相比，IOIP组织培养的细胞生长速度较快。随着地塞米松浓度的增加，成纤维细胞的增生值（A值）逐渐下降，0、1×10-3、1×10-4、1×10-6和1×10-6mol／L地塞米松组成纤维细胞增生值的总体比较差异有统计学意义（F浓度=36．27，P=0．00），随着地塞米松作用时间的延长，成纤维细胞增生值逐渐下降，总体比较差异有统计学意义（F时间=3．69，P=0．00）。无地塞米松培养组培养24、48和72h，成纤维细胞中比较ICAM-1表达量分别为0．298±0．008、0．312±0．003和0．319±0．011，表现为逐渐升高的趋势，而不同浓度地塞米松作用后随着时间延长ICAM-1表达量均逐渐降低，总体比较差异有统计学意义（F时间=3．11，P=0．00），随着地塞米松浓度的增加，ICAM-1表达量均逐渐下降，总体比较差异有统计学意义（F浓度=75．17，P=0．00）。结论IOIP的发生和发展可能与眼眶成纤维细胞中ICAM-1表达增强有关，地塞米松可能通过下调细胞中ICAM-1的表达而抑制眼眶成纤维细胞的增生，从而发挥抗炎和治疗作用。
Background Idiopathic orbital inflammatory pseudotumor （IOIP） but its etiology is still unclear, so the effect of glucocorticoid treatment is unsatisfied is a common orbital disease, Objective This study was to investigate the effects of dexamcthasone on orbital fibroblasts from IOIP patients and explore the action machanism. Methods Six pieces of IOIP tissues from 6 IOIP patients and 3 pieces of normal orbital connective tissues from lacrimal gland prolapse patients were obtained during the surgery in Beijing Tongren Hospital from November 2011 to January 2012. The orbital fibroblasts were cultured using explant culture method. The morphology of the cells were observed under the optical microscope,and biomarks of the cells were detected by immunochemistry. The growth and proliferation of the cells were assayed using WST-8. The expression of ICAM-1 in the cells in both the control group and the IOIP group was detected by immunochemistry. The fibroblasts were incubated in 96-well plates,and different concentrations of dexamethasone （0,1 ×10--3 , 1 ×10-4 , 1 ×10-5 and 1×10-6 mol/L） were respectively added into the medium for 24,48 and 72 hours,and then the proliferation of the cells was detected by WST-8 assay. The contents of ICAM-1 in different concentrations of dexamethasone groups were assayed by ELISA. Results The characteristics of the cells were similar between the control group and the IOIP group with the spindle shape and long protructions. The cells showed the positive response for vimentin and absent response for desmin, S-100, cytokeratin （CK）. Compared with the control group,the growth speed of fibroblasts was fast in the IOIP group. The proliferative values of the cells （absorbancy） were gradually reduced with the increase of dexamethasone concentrations （F tration = 36. 27, P = 0. 00 ） and the lapse of acting time （ Ftime = 3. 69, P = 0.00）. In cultured cells without dexamethasone for 24,48 and 72 hours,the mean expression levels of ICAM-1 were 0. 298±0. 008,0. 312±0. 003 and 0. 319±0. 011 , showing a gradually increasing trend. However,the expression of ICAM-1 was gradually reduced with the increases of concentrations and the lapse of acting time of dexamethasone （ F concentration= 75. 17, P = 0. 00 ; Ftime = 3. 11, P = 0. 00 ）. Conclusions Occurrence and development of IOIP is probably associated with the over-expression of ICAM-1 in orbital fibroblasts. Dexamethasone plays anti-inflammation and treating effects on IOIP by down-regulating the expression of ICAM-1 and inhibiting the proliferation of orbital fibroblasts.
出处
《中华实验眼科杂志》
CAS
CSCD
北大核心
2015年第11期1004-1008,共5页
Chinese Journal Of Experimental Ophthalmology
基金
国家自然科学基金项目（81170875）
北京市卫生系统高层次卫生技术人才培养计划项目（2011-3-041）
