Milenkovic Milutin, De Deken Xavier, Jin Ling, De Felice Mario, Di Lauro Roberto, Dumont Jacques E, Corvilain Bernard, Miot Francoise
IRIBHM, Erasme University Hospital, Free University of Brussels, Route de Lennik, 808, 1070 Brussels, Belgium.
J Endocrinol. 2007 Mar;192(3):615-26. doi: 10.1677/JOE-06-0003.
In the thyroid, H(2)O(2) is produced at the apical pole of thyrocytes by one or two NADPH oxidases (NOX), Duox1/2 proteins. The onset of Duox expression was analysed by immunohistochemistry in the developing mouse thyroid in parallel with thyroglobulin (Tg) iodination and the expression of other thyroid differentiation markers. Duox proteins were found at embryonic day (E) 15.5 and were mainly localised at the apical pole of thyrocytes. Tg was detected 1 day before (E14.5) and Tg iodination was concomitant with the expression of both Duox and Na(+)/I(-) symporter (NIS; E15.5). The role of TSH in regulating Duox expression and H(2)O(2) accumulation was evaluated in thyroids of adult mice with reduced (Tshr(hyt/hyt) or mice treated with thyroxine) or increased (methimazole or perchlorate treatment) TSH/Tshr activity. In mice with suppressed TSH/Tshr activity, Duox expression was only partially decreased when compared with wild-type, as observed by western blot. In Tshr(hyt/hyt) strain, Duox was still expressed at the apical pole and H(2)O(2) measurements were normal. On the other hand, chronic TSH stimulation of the gland led to a decrease of H(2)O(2) measurements without affecting Duox expression. The onset of Duox protein expression is compatible with their proposed function in thyroid hormone synthesis and it can be considered as a functional marker of the developing thyroid. However, Duox expression in adult is much less regulated by TSH than NIS and thyroperoxidase. It is not always correlated with the overall thyroid H(2)O(2) accumulation, highlighting the importance of additional regulatory mechanisms which control either the production or H(2)O(2) degradation.
在甲状腺中,甲状腺细胞顶端的过氧化氢(H₂O₂)由一种或两种烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶(NOX)即双氧化酶1/2(Duox1/2)蛋白产生。通过免疫组织化学方法,在发育中的小鼠甲状腺中,平行分析双氧化酶(Duox)的表达起始情况,同时检测甲状腺球蛋白(Tg)碘化及其他甲状腺分化标志物的表达。在胚胎第15.5天(E15.5)发现了Duox蛋白,其主要定位于甲状腺细胞的顶端。在E14.5前一天检测到了Tg,而Tg碘化与Duox和钠/碘同向转运体(NIS;E15.5)的表达同时出现。通过降低(促甲状腺激素受体基因敲除小鼠(Tshr(hyt/hyt))或用甲状腺素处理的小鼠)或增加(用甲巯咪唑或高氯酸盐处理)促甲状腺激素(TSH)/促甲状腺激素受体(Tshr)活性,评估TSH在调节Duox表达和H₂O₂积累中的作用。通过蛋白质免疫印迹法观察到,在TSH/Tshr活性受到抑制的小鼠中,与野生型相比,Duox表达仅部分降低。在Tshr(hyt/hyt)品系中,Duox仍在顶端表达,且H₂O₂测量结果正常。另一方面,对腺体进行慢性TSH刺激导致H₂O₂测量值降低,但不影响Duox表达。Duox蛋白表达的起始情况与其在甲状腺激素合成中所提出的功能相符,可被视为发育中甲状腺的一个功能标志物。然而,与NIS和甲状腺过氧化物酶相比,成年期Duox表达受TSH的调节要少得多。它并不总是与甲状腺整体H₂O₂积累相关,这突出了控制H₂O₂产生或降解的其他调节机制的重要性。