目的观察NOD.H-2h4小鼠自身免疫性甲状腺炎时甲状腺内细胞凋亡和凋亡相关蛋白Fas、Fas-L和FLIP的表达情况,研究细胞凋亡在自身免疫性甲状腺炎中的作用。方法7~8周龄的NOD.H-2h4小鼠,饲以0.05%NaI溶液作为实验组,对照组小鼠则饲以自来水。在给碘后8,16,32周处死小鼠,HE染色观察甲状腺的炎症程度,TUNEL方法观察甲状腺内细胞凋亡情况,免疫组化方法观察凋亡相关蛋白Fas、Fas配体(Fas-L)和FLICE抑制蛋白(FLIP)的表达情况。结果实验组小鼠出现自身免疫性甲状腺炎,对照组无甲状腺炎产生。甲状腺炎时,TUNEL染色阳性细胞主要集中在浸润淋巴细胞中,甲状腺滤泡上皮细胞罕见凋亡;Fas、Fas-L和FLIP主要表达在甲状腺内的浸润淋巴细胞上;部分甲状腺滤泡上皮细胞表达Fas,而Fas-L和FLIP在甲状腺滤泡上皮细胞未见明显表达。结论NOD.H-2h4小鼠自身免疫性甲状腺炎时甲状腺细胞无明显凋亡,提示细胞凋亡机制未参与甲状腺炎的发病。甲状腺内浸润淋巴细胞虽然表达Fas但并未出现明显凋亡,这可能与其同时表达抗凋亡蛋白FLIP有关。
Objective To observe the status of cell apoptosis and the expression of apoptosis related protein Fas, Fas-L and FLIP in au- toimmune thyroiditis of NOD.H-2h4 mice, and investigate the role of apoptosis in autoirmnune thyroiditis. Methods 7 to 8 week-old NOD.H- 2h4 mice of experimental group were administrated with 0.05% NaI solution to induce autoimmune thyroiditis, the mice of control group received water. Mice in both groups were sacrificed at the time point of 8,16 and 32 weeks. The degree of autoimmune thyroiditis was evaluated by HE staining. Apoptosis of thyrocytes and infiltrating lymphocytes was detected by TUNEL method, lmmunohis/ochemistry staining was conducted to analyze the expression of apoptosis related proteins Fas, Fas-L and FLIP. Results Apparent autoimmune thyroiditis were found in mice received 0.05% NaI solution. TUNEL positive cells were mainly distributed in infdtrating lymphocytes and scarcely found in thyroid follicular cells. Fas, Fas-L and FLIP expressed strong in infiltrating lymphocytes. Part of the thyroid follicular cells expressed Fas protein, and no expressed Fas-L and FLIP protein was found in thyroid follicular cells. Conclusion No obvious apoptosis of thyroid follicular cells was found in autoinunune thyroiditis NOD.H-2h4 mice. Apoptosis does not involve in the mechanism of autoimmune thyroidifis. The simultaneous expression of anti-apoptosis protein FLIP and Fas protein in infiltrating lymphocytes may relate to the less apoptosis in thyroiditis.