瘦素基因在甲状腺相关眼病眶脂肪及体外分化眶脂肪细胞中的表达

Expression of leptin gene in thyroid associated ophthalmopathy orbital adipose and differentiated adipocytes in vitro

目的探讨瘦素基因在甲状腺相关眼病(thyroid associated ophthalmopathy,TAO)患者和正常人眼眶脂肪组织和体外诱导分化的眼眶脂肪细胞中的表达及意义。方法Trizol法提取8例(8眼)TAO患者和13例(13眼)正常人眼眶脂肪组织总RNA,采用实时荧光定量PCR检测瘦素mRNA的表达。体外培养TAO患者与正常人眼眶前脂肪成纤维细胞并进行诱导分化,实时荧光定量PCR检测分化后脂肪细胞中瘦素mRNA的表达。结果 TAO患者眼眶脂肪组织中瘦素mRNA的相对表达量为2.05±0.31(已排除年龄、性别和身体质量指数可能的影响),正常人眼眶脂肪组织中瘦素mRNA的相对表达量为0.69±0.24,两者相比差异有统计学意义(P<0.05)。体外培养的TAO患者和正常人眼眶前脂肪成纤维细胞在诱导分化刺激下,均能分化为脂肪细胞;TAO患者分化第10天眼眶脂肪细胞的瘦素mRNA的相对表达量为1.55±0.44,正常人分化第10天眼眶脂肪细胞的瘦素mRNA的相对表达量为0.66±0.21,两者相比差异有统计学意义(P<0.05)。结论瘦素基因在TAO患者眼眶脂肪组织和体外诱导分化的眼眶脂肪细胞中的表达均明显高于正常人,提示瘦素基因可能参与了TAO的发病机制。

Objective To investigate the expression and significance of leptin gene in orbital adipose and differentiated adipocytes in vitro of thyroid associated ophthalmopathy （TAO） patients and normal humans. Methods Total RNA was extracted from orbital adipose of 8 patients （8 eyes） and 13 normal humans （13 eyes） by Trizol method, and the expression of leptin mRNA was detected by real-time fluorescent quantitative PCR. Orbital preadipocytes of patients and normal humans were induced to dif- ferentiated adipocytes in vitro, and the expression of leptin mRNA in these cells was detected by real-time fluorescent quantitative PCR. Results The relative expressions of leptin mRNA in orbital adipose of patients and normal humans were 2.05 ± 0.31, 0.69 ± 0.24, respectively, the difference between them was significant （ P 〈 0.05 ）. The orbital preadipocytes of patients and normal humans could differentiate to adipocytes in vitro. The relative expressions of leptin mRNA in those adipocytes at differentiated 10 days were 1.55 ±0.44,0.66±0.21 ,respectively,the difference between them was significant（P 〈 0.05 ）. Conclusion The expressions of leptin gene in orbital adipose and differentiated adipocytes in vitro of TAO patients are higher than those of normal hu- mans, which suggest that leptin may be involved in the pathogenesis of TAO.