摘要
目的观察脂多糖(LPS)对甲状腺相关眼病(TAO)眼眶成纤维细胞环氧合酶-2(COX-2)表达及前列腺素E2(PGE2)分泌的影响。方法眼眶组织分别取自3例3眼因眼球萎缩行义眼台植入(正常对照组)和4例4眼因TAO行开眶减压术(TAO组)的患者。细胞原代培养采用组织块法。将体外培养的2组眼眶成纤维细胞,分别采用0、10、100、1000μg/L的LPS作用24h,或1000μg/LLPS分别作用0、4、8、12、24h。半定量逆转录聚合酶链反应法(RT-PCR)检测各组COX-2mRNA的表达,ELISA法检测上清液中PGE2的表达。结果组织块培养法第3天即有成纤维样细胞自组织块边缘游出,细胞呈长梭形和多角形。10、100、1000μg/LLPS作用后,与未用LPS刺激时相比,正常对照组和TAO组眼眶成纤维细胞COX-2mRNA的表达均增强,PGE2的分泌增加。与正常对照组相比,1000μg/LLPS作用时间相同时,TAO组COX-2mRNA表达及PGE2的分泌增加更为明显(P<0.05)。结论 COX-2通路可能参与了活动期TAO的炎症过程;在疾病的活动期,采用抑制COX-2的治疗,可减轻局部炎症反应,可能对活动期TAO的治疗有效。
Background The study of the pathogenesis mechanism of thyroid-associated ophthalmopathy （TAO） is a hot research topic.Experiment showed that the inflammation is involved in the pathophysiology procedure of TAO.Objective The aim of this study was to investigate the effects of lipopolysaccharide （LPS） on the expression of cyclooxygenase-2 （COX-2） and secretion of prostaglandin E2（PGE2） in orbital fibroblasts of TAO.Methods Orbital adipose tissues were obtained from 3 eyes of 3 patients with eyeball atrophy due to ocular injury （normal control group） or 4 eyes of 4 patients with TAO （TAO group） during the surgery.The orbital adipose tissue was cultured with explant method to obtain the orbital fibroblasts.Cultured orbital fibroblasts were incubated in culture medium containing different concentrations of LPS（0,10,100,1 000 μg/L）for 24 hours.In additional,obtained orbital fibroblasts was stimulated in 1 000 μg/L LPS for 0 hour,4 hours,8 hours,12 hours,24 hours respectively.The expression of COX-2 mRNA in orbital fibroblasts in both groups were semiquantatively analyzed by reverse transcription polymerase chain reaction （RT-PCR）,and the secretion level of PGE2 in the supernatant was detected by ELISA.The oral informed consent was obtained from the patients prior to this medical process.Results Orbital fibroblasts emigrated from tissue explants after 3 days of culture and showed the fusiformis and polygonal in shape.The expression of COX-2 mRNA was significantly increased in the orbital fibroblasts in TAO group compared with normal control group after 10,100,1 000 μg/L LPS action（all P0.05） and 4,8,12 and 24 hours after action of 1 000 μg/L LPS （all P0.05）.The secretion level of PGE2 was considerably enhanced in 10,100,1 000 μg/L LPS group compared with control group and different time points after action of 1 000 μg/L LPS （all P0.05）.Conclusion COX-2 pathway is involved in the inflammatory mechanism of active TAO.To block the expression of COX-2 and attenuate local inf
出处
《眼科研究》
CSCD
北大核心
2010年第7期596-600,共5页
Chinese Ophthalmic Research
基金
湖南省科技厅基金项目(2008JT3002)
