TNF-α对甲状腺相关眼病分化后眼眶脂肪细胞瘦素mRNA表达的影响

Effect of TNF-α on the expression of leptin mRNA in orbital differentiated adipocytes of thyroid associated ophthalmopathy

目的探讨肿瘤坏死因子-α(TNF-α)对体外培养的甲状腺相关眼病(TAO)分化后眼眶脂肪细胞瘦素mRNA表达的影响。方法取TAO患者行眼眶减压术的眼眶脂肪组织,体外培养眼眶前脂肪细胞,诱导分化,采用不同质量浓度、不同作用时间的TNF-α处理分化后的脂肪细胞,实时荧光定量PCR检测瘦素mRNA的表达。结果体外培养的眼眶前脂肪细胞在诱导分化刺激下,分化为脂肪细胞;0.5～50ng/mL的TNF-α作用24h,瘦素mRNA表达随质量浓度的增大而下降,呈量效依赖性;5ng/mL的TNF-α作用12～48h能下调瘦素mRNA的表达,呈时效依赖性;与对照组相比,差异均有统计学意义(P<0.05)。结论TNF-α对TAO分化后眼眶脂肪细胞的瘦素mRNA表达有下调作用。

Objective Thyroid associated ophthalmopathy（TAO） is an autoimmunity disease with hypertrophy of ocular muscle and increase of ocular fat. It is known that many factors are involved in its pathogenesis,but influence of leptin in genesis and development of TAO remain unclear. This study was curried out to investigate the effect of TNF-α on the expression of leptin mRNA in orbital differentiated adipocytes of thyroid associated ophthalmopathy （TAO） subjects. Methods Preadipocytes from orbital tissues of TAO patients after orbital decompression were cultured and committed to differentiate. The differentiated orbital adipocytes were incubated with 0. 5,5,50 ng/mL of TNF-α for 24 hours or 5 ng/mL TNF-α for 0, 12,24,48 hours. The expression of leptin mRNA in the differentiated adipocytes was detected by real-time fluorescent quantitative PCR. Results Induced preadipocytes differentiated into the cells with morphological feature of mature adipocyte under the appropriate in vitro condition with the red staining for oil red. After treated with 0. 5 - 50 mg/L TNF-α, the expression of leptin mRNA in the differentiated adipocytes was significantly suppressed in comparison with control group （ F = 10. 401, P 〈 0. 05 ）. The expression of leptin mRNA in the differentiated adipocytes was significantly decreased in 12,48 and 72 hours after 5 ng/mL of TNF-α treatment in comparison with before treatment （ F = 28. 834, P 〈 0. 01 ）. Conclusion TNF-α can suppress the expression of leptin mRNA in the differentiated adipocytes in a dose- and time-dependent manner.