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可溶性CD40L促进甲状腺相关性眼病患者眼眶成纤维细胞增殖和透明质酸合成酶表达
曾程程,魏锐利*,牟旆,王秋红
0
(第二军医大学长征医院眼科, 上海 200433
*通信作者)
摘要:
目的 探讨可溶性CD40L(sCD40L)对甲状腺相关性眼病(TAO)患者眼眶成纤维细胞(OFs)增殖和3种透明质酸合成酶(HAS) mRNA表达水平的影响,初步探讨sCD40L在TAO发病中的作用。 目的 取5例TAO患者和3例正常对照标本原代培养OFs,采用MTS比色法检测终质量浓度6.25、12.5、25、50、100、200 ng/mL的sCD40L作用48 h后对不同来源OFs增殖的影响;实时荧光定量PCR技术检测终质量浓度12.5、25、50、100 ng/mL及作用3、6、12、24 h后的sCD40L对不同来源OFs的3种HAS基因(HAS1~3)表达水平的影响。 结果 25 ng/mL及以上浓度的sCD40L作用48 h后对TAO患者OFs有促增殖作用(P<0.05);而50 ng/mL及以上浓度的sCD40L作用48 h后才能对正常对照OFs有促增殖作用(P<0.05),且作用较弱。TAO患者OFs中HAS3 mRNA的合成在sCD40L的刺激下增加,并且呈现出浓度和时间依赖的趋势。 结论 sCD40L能够促进TAO患者OFs的生长以及HAS3基因的表达,提示sCD40L在TAO的病理过程中起一定作用。
关键词:  可溶性CD40配体  Graves眼病  眼眶成纤维细胞  透明质酸合成酶
DOI:10.3724/SP.J.1008.2015.01138
投稿时间:2015-05-07修订日期:2015-09-06
基金项目:国家自然科学基金(81371056).
Soluble CD40L promotes proliferation of orbital fibroblasts and expression of hyaluronan synthase in patients with thyroid associated ophthalmopathy
ZENG Cheng-cheng,WEI Rui-li*,MOU Pei,WANG Qiu-hong
(Department of Ophthalmology, Changzheng Hospital, Second Military Medical University, Shanghai 200433, China
*Corresponding author.)
Abstract:
Objective To investigate the effects of soluble CD40L (sCD40L) on proliferation of orbital fibroblasts (OFs) and the expression of three types of hyaluronan synthase (HAS) in vitro, so as to explore the role of sCD40L in the pathogenesis of thyroid-associated ophthalmopathy (TAO). Methods OFs obtained from 5 patients with TAO and 3 normal controls were primarily culutred. The effect of different concentrations of sCD40L (6.25,12.5,25,50,100 and 200 ng/mL) on proliferation of OFs of different sources were examined by MTS after 48 h exposure. OFs were also cultured with different concentrations of sCD40L (12.5, 25, 50 and 100 ng/mL) for 3,6,12 and 24 h, and then the expression levels of HAS 1-3 mRNA were determined by real-time RT-PCR. Results Treatment with sCD40L at concentrations higher than 25 ng/mL for 48 h obviously promoted the proliferation of OFs in patients with TAO (P<0.05). In contrast, treatment with sCD40L only at concentrations higher than 50 ng/mL for 48 h could promote proliferation of OFs from normal control, and the effect was comparatively weak. HAS3 mRNA expression of OFs in TAO patients was increased after exposed to sCD40L (P<0.05), and the increase was in a concentration- and time-dependent manner. Conclusion sCD40L can promote the proliferation of OFs and expression of HAS3 mRNA in patients with TAO, which implies that sCD40L plays an important role in the pathogenesis of TAO.
Key words:  soluble CD40 ligand  Graves ophthalmopathy  orbital fibroblasts  hyaluronan synthase