摘要
目的研究肿瘤坏死因子α(TNFα)在有或无促甲状腺素(TSH)存在时,对甲状腺培养细胞甲状腺球蛋白(TG)、甲状腺过氧化物酶(TPO)基因转录的影响。方法运用细胞原代培养技术和NorthernBlot分子杂交技术,观察不同剂量的TNFα对甲状腺细胞TG、TPOmRNA表达的调节作用。结果(1)TNFα在0.01U/ml时能够明显降低甲状腺细胞基础状况下TG、TPOmRNA的水平,当TNFα处于1~100U/ml时,TG、TPOmRNA的水平几乎检测不出;(2)TSH在10mU/ml时显著提高甲状腺细胞表达TGmRNA,TNFα在0.01~1U/ml剂量范围内对TSH的该作用无明显影响,但当TNFα浓度在100~1000U/ml时,对TSH的该作用表现为抑制效应。(3)TSH在10mU/ml时显著提高甲状腺细胞表达TPOmRNA,TNFα在0.01U/ml剂量范围内对TSH的该作用无明显影响,但当TNFα浓度在1~1000U/ml时,对TSH的该作用表现为抑制效应。结论TNFα能够在基因转录水平上抑制甲状腺细胞基础合成TG、TPOmRNA,并且TNFα在较高水平时能够拮抗TSH促进TG、TPOmRNA表达的作?
Objective To study the effects of TNFα with or without TSH on the thyroglobulin (TG) and thyroid peroxidase (TPO) gene mRNA levels. Methods Thyrocytes were isolated from thyroid tissues of patients with Graves' disease and TNFα at different doses were added in the absence or presence of bovine TSH. Total RNA was extracted from thyrocytes using a single step method and it was electrophoresed and transferred to nylon filter. The filter was hybridized to TG, TPO and and GAPDH probes respectively and subjected to autoradiography. Results (1) Basal TG and TPO mRNA levels of thyrocytes were significantly decreased by TNFα of 0.01U/ml and were inhibited to undetectable level by TNFα of 1~100U/ml; (2) TNFα in the concertration of 0.01U/ml~1U/ml did not affect TSH induced TG mRNA levels while at the dose of 100~1000U/ml TNFα significantly inhibited TSH induced TG gene transcription; (3) TNFα in the concentration of0.01U/mldidnotaffectTSH induced TPO mRNA levels, but TNFα at 1~1000U/ml significantly decreased TSH induced TPO mRNA. Conclusion TNFα directly affects TG and TPO gene transcription in cultured human thyrocytes without TSH and TNFα does not affect TSH stimulated TG, TPO mRNA levels at lower doses while decreases these levels at higher doses.
出处
《中华内分泌代谢杂志》
CAS
CSCD
北大核心
1999年第1期31-34,共4页
Chinese Journal of Endocrinology and Metabolism