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目的:通过建立实验性自身免疫性甲状腺炎(experimentalautoimmunethyroiditis,EAT)动物模型的基础上,探索通过基因转染方法,以纠正EAT大鼠免疫功能异常状态。方法:选用雌性Wistar大鼠建立EAT动物模型。成模大鼠分为4组:A组5只(PBS+PLL),B组5只(pORF质粒+PLL)、C组10只(pORFmIL10质粒+PLL)及D组5只(pORFmIL10质粒+MEM)。使用放射免疫方法测定TgAb,TmAb滴度。采用Charreive法计算甲状腺内淋巴细胞浸润指数。脾脏淋巴细胞培养,采用犤3H犦-TdR标记方法测定cpm值并计算淋巴细胞增殖指数。结果:C组与A,B,D组比较,第4,6,8周TgAb,TmAb滴度显著降低(P均<0.001,F=42.66,32.65,9.66;F=22.25,81.35,14.84)。C组甲状腺淋巴细胞浸润指数为1.10±0.18,显著低于其他各组(F=4.39,P<0.01)。此外,淋巴细胞增殖试验结果显示,C组PTg刺激的淋巴细胞增殖指数显著低于A,B组(F=2.78,P<0.05)。结论:通过大鼠甲状腺组织内局部注射pORFmIL10质粒使甲状腺滤泡上皮细胞表达白细胞介素10基因,并且能够清除甲状腺内浸润的淋巴细胞,降低自身抗体水平和针对抗原反应的T淋巴细胞增殖反应。多聚赖氨酸可以延长目的基因表达时间,更有效地治疗EAT。
OBJECTIVE: To establish an experimental animal model of experimental autoimmune thyroiditis (EAT) to explore the possibility of correcting immune dysfunction in EAT rats by gene transfection. Methods: Female Wistar rats were selected to establish EAT animal model. Rats in model group were divided into 4 groups: 5 in group A (PBS + PLL), 5 in group B (pORF plasmid + PLL), 10 in group C (pORFmIL10 plasmid + PLL) and 5 in group D (pORFmIL10 plasmid + MEM ). TgAb, TmAb titers were determined using radioimmunoassay. Charreive method was used to calculate thyroid lymphocyte infiltration index. Splenic lymphocytes were cultured. The cpm value was measured by 犤 3H-TdR labeling method and the lymphocyte proliferation index was calculated. Results: The titers of TgAb and TmAb in group C were significantly lower than those in groups A, B and D (P <0.001, F = 42.66, 32.65 and 9.66; F = 22.25, 81.55 and 14.84) at weeks 4, 6 and 8, respectively. The index of thyroid lymphocyte infiltration in group C was 1.10 ± 0.18, which was significantly lower than that in other groups (F = 4.39, P <0.01). In addition, lymphocyte proliferation test results showed that PTg-stimulated lymphocyte proliferation index of group C was significantly lower than that of group A and B (F = 2.78, P <0.05). CONCLUSION: The injection of pORFmIL10 plasmid into rat thyroid tissue allows the expression of interleukin-10 gene in thyroid follicular epithelial cells. It can also eliminate infiltrating lymphocytes in the thyroid gland and reduce the level of autoantibodies and the proliferation of T lymphocytes against antigen reactions. Polylysine can prolong the target gene expression time and treat EAT more effectively.