甲状腺相关眼病及正常人眼眶成纤维细胞与前脂肪细胞培养及分化诱导  

Culture and differentiation of fibroblasts and preadipocyte fibroblasts derived from thyroid associated ophthalmopathy

作　　者：张学进[1] 罗清礼[1] 何为民[1] 刘春玲[1] 李涛[1] 

机构地区：[1]四川大学华西医院眼科,成都610041

出　　处：《眼科研究》2007年第2期81-85,共5页Chinese Ophthalmic Research

基　　金：国家自然科学基金资助(30371518)

摘　　要：目的研究不同浓度3-异丁基-1-甲基黄嘌呤(IBMX)、胰岛素、地塞米松对培养的甲状腺相关眼病(TAO)及正常人眼眶成纤维细胞与前脂肪细胞的诱导作用。方法原代培养13个TAO与7个正常人眼眶成纤维细胞与前脂肪细胞;透射电镜检查超微结构;油红O染色,光度法检测不同浓度地塞米松、胰岛素、IBMX对脂肪细胞转化率和细胞内脂肪含量的影响。结果培养了TAO与正常人眼眶成纤维细胞与前脂肪细胞,并诱导前脂肪细胞向脂肪细胞转化,获得最佳的诱导浓度地塞米松0.1μmol/L、胰岛素10μg/ml、IBMX0.25mmol/L。结论TAO患者与正常人眼眶成纤维细胞和前脂肪细胞在形态学上未见明显差异,但前脂肪细胞能诱导分化为脂肪细胞,眼眶前脂肪细胞在TAO的发病中可能起重要作用。Objective The orbital fatty tissue plays an important role in pathogenesis of thyroid associated ophthalmopathy （TAO） by elevating intraocular pressure and secreting many types of fatty cell factors and growth factors etc.. In the current study, we investigated the revulsive effects of dexamethasone, insulin and 3-isobutyl-1-1 methylxanthine （IBMX） on the fibroblasts and preadipocyte fibroblasts derived from TAO. Methods Orbital fibroblasts and preadipocytes obtained from 13 patients with TAO and 7 normal corpse eyes were primarily cultured and the ultrastructure of cultured cells was observed by transmission electron microscopy. The effects of different concentrations of dexamethasone,insulin and IBMX on the preadipocyte differentiation were compared by spectrophotometer. Results The orbital fibroblasts and preadipocytes were successfully cultured. Both cells exhibited stellate shape, showing brown positive reaction in cytoplasm for vimtin. Fibroblasts and preadipocyte fibroblasts had abundance of mitochondria and ergastoplasm in the cytoplasm with a similar ultrastructure. The preadipocytes were induced into adipocytes,displaying some inclusions in the cytoplasm 1 week to 2 weeks after differentiation protocol, and some inclusions amalgamated and enlarged with protocol continued. The optimal revulsive concentrations of dexamethasone,insulin and IBMX were 0. 1 μmol/L, 10 μg/ml and 0.25 mmob/L, respectively. Conclusion There are no significant morphological differences between the orbital fibroblasts and preadioocytes from patients and normal subjects. Preadipocytes can differentiate into adipocytes. Orbital preadipocytes may play an important role in the pathogenesis of TAO.

关 键 词：甲状腺相关眼病 前脂肪细胞 细胞培养 分化 

分 类 号：R771.3[医药卫生—眼科]