脂多糖诱导炎症反应对甲状腺相关眼病眼眶前脂肪细胞分化的影响  

Effects of LPS-induced inflammation on differentiation of orbital pre-adipocytes in thyroid-associated ophthalmopathy

作　　者：易文殊[1] 许雪亮[1] 

机构地区：[1]中南大学湘雅医院眼科易文殊为博士研究生,现在武警福建总队医院眼科,长沙410008

出　　处：《中华眼科杂志》2011年第2期156-161,共6页Chinese Journal of Ophthalmology

基　　金：湖南省科技厅基金

摘　　要：目的观察脂多糖诱导炎症反应对甲状腺相关眼病（TAO）眼眶前脂肪细胞分化的影响，探讨TAO眼眶脂肪增殖的发病机制。方法实验研究。眼眶组织取TAO行开眶减压术的患者。将体外培养的TAO眼眶成纤维细胞，分为A组与B组，A组采用1mg／L的脂多糖作用8h，然后常规用诱导分化液I和Ⅱ诱导直至分化结束，B组仅用诱导分化液I和Ⅱ诱导至分化结束。采用油红O染色法测定分化后脂肪细胞相对含量，逆转录聚合酶链反应法与Western blot法检测环氧化酶2（COX2）与过氧化物酶体增殖物激活受体γ（PPARγ）mRNA与蛋白的表达，酶联免疫吸附实验检测上清液中前列腺素E2（PGE2）的表达。两样本均数比较采用t检验，多样本比较采用单因素方差分析，两两比较采用SNK检验。结果分化后油红O染色显示两组细胞形态上无明显差异。A组细胞吸光度（A）值（1．02±0．08）较B组（0．74±0．06）明显升高，差异具有统计学意义（t=8．502，P=0．000）。A组细胞分化后PPARγ mRNA（1．74±0．19）与蛋白（0．47±0．04）的表达分别较分化前PPARγ mRNA（0．30±0．07）与蛋白（0．08±0．02）的表达明显增强（P〈0．05）；B组细胞分化后PPARγmRNA（1．15±0．18）与蛋白（0．35±0．03）的表达分别较分化前PPARγ mRNA（0．13±0．04）与蛋白（0．03±0．01）的表达明显增强（P〈0．05）；A组细胞分化后COX2 mRNA（0．28±0．07）与蛋白（0．24±O．03）的表达分别较分化前COX2mRNA（1．54±0．10）与蛋白（O．49±0．03）的表达明显减弱（P〈0．05）；B组细胞分化后COX2mRNA的表达（0．08±0．03）较分化前（0．19±0．07）明显减弱（P〈0．05），而COX2蛋白的表达（0．01±0．01）与分化前（0．03±0．01）比较差异无统计学意义（P〉0．05）。分化后A组细胞上清液中PGE2分泌水平（208．43±15．06）ng／L较分化前（898．75±21．09）Objective To investigate the effects of lipopolysaccharide （LPS） -induced inflammation on the differentiation of orbital pre-adipocytes in thyroid-associated ophthalmopathy （TAO） and to explore the mechanism of orbital adipose proliferation in TAO. Methods Orbital adipose tissues were obtained from patients with TAO undergoing orbital decompression surgery. The orbital fibroblasts cultured from orbital adipose tissues were divided into group A and group B. In group A, orbital pre-adipocytes were incubated in culture medium containing 1 mg/L LPS for 8 hours before stimulated to differentiate into mature adipocyteswith Differentiation Medium I and 11. No LPS or other intervention was used in group B before induced to differentiate into mature adipocyte with Differentiation MediumⅠ and Ⅱ. Intracellular fat accumulation in differentiated adipocytes was determined by oil red O staining and the expression of cyclooxygenase2 （COX2） and peroxisome proliferators activated receptor γ （PPARγ） mRNA of both groups were detected by RT-PCR. Protein expression of COX2 and PPARγ in both groups was detected by Western-blot and the secretion of PGE2 in the supernatant was detected by ELISA. Results COX2 expression and secretion of PGE2 in differentiated ceils of both groups were significantly decreased compared with pre-differentiation （ P 〈 0. 05 ）, while PPARγmRNA and protein expression enhanced significantly （ P 〈 0. 05 ）. COX2 mRNA and protein expression and secretion of PGE2 of pre-differentiation cells in group A was significantly increased compared with group B （ P 〈 0. 05 ）, while PPARγmRNA and protein expression in group A was also stronger than those in the group B （ P 〈 0. 05 ）. COX2 and PPARγ mRNA and protein expression and secretion of PGE2 of differentiated cells in group A were greater than those in group B （ P 〈 0. 05 ）. Conclusion LPS can induce inflammatory response of orbital preadipocytes in TAO and enhance the expression of COX2, PPARγ and PGE2. The expre

关 键 词：脂多糖类 GRAVES眼病 环氧化酶2 PPARγ 细胞分化 

分 类 号：R581.1[医药卫生—内分泌] R771.3[医药卫生—内科学]