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。方法 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、200ng/mL的sCD40L作用48h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、100ng/mL及作用3、6、12、24h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 25ng/mL及以上浓度的sCD40L作用48h后对TAO患者OFs有促增殖作用（P〈0.05）;而50ng/mL及以上浓度的sCD40L作用48h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HAS3mRNA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。
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 sCD40 Lin the pathogenesis of thyroid-associated ophthalmopathy（TAO）.Methods OFs obtained from 5patients with TAO and 3normal controls were primarily culutred.The effect of different concentrations of sCD40L（6.25,12.5,25,50,100 and 200ng/mL）on proliferation of OFs of different sources were examined by MTS after 48 hexposure.OFs were also cultured with different concentrations of sCD40L（12.5,25,50 and 100ng/mL）for 3,6,12 and 24h,and then the expression levels of HAS1-3mRNA were determined by real-time RT-PCR.Results Treatment with sCD40 Lat concentrations higher than 25ng/mL for 48 h obviously promoted the proliferation of OFs in patients with TAO（P〈0.05）.In contrast,treatment with sCD40 Lonly at concentrations higher than 50ng/mL for 48 hcould 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 sCD40 Lcan promote the proliferation of OFs and expression of HAS3 mRNA in patients with TAO,which implies that sCD40 Lplays an important role in the pathogenesis of TAO.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2015年第10期1138-1142,共5页
Academic Journal of Second Military Medical University
基金
国家自然科学基金(81371056)~~
