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目的研究甲宁方对甲状腺刺激抗体(TSAb)作用下Fisher大鼠甲状腺细胞系(FRTL细胞)增殖影响和诱导凋亡作用。方法将甲状腺功能亢进(Graves病)患者血清经聚乙二醇4000处理获得TSAb的粗提物作用于FRTL细胞,将甲宁方含药血清分为低、中、高浓度组,并设甲亢灵片对照组,与FRTL细胞共培养后,用MTT法检测细胞生长状况,用原位杂交技术检测bcl-2、baxmRNA表达。结果TSAb阳性血清粗提物促进FRTL细胞生长,不同浓度甲宁方组与FRTL细胞共培养后,细胞生长受抑制,呈浓度依赖性,高剂量时作用显著(P<0.01);甲宁方各组bcl-2阳性表达率降低,中、高浓度甲宁方组bax阳性表达率升高;甲宁方降低bcl-2/bax比值,从中浓度至高浓度作用逐渐增强。结论甲宁方可以有效抑制TSAb作用下FRTL细胞增殖,诱导其凋亡,这可能是该方治疗Graves病的机理之一。
Objective To investigate the effect of Jianing prescription on the proliferation of Fisher rat thyroid cell line (FRTL) under the action of thyroid stimulating antibody (TSAb) and induce apoptosis. Methods The serum of patients with hyperthyroidism (Graves’ disease) was treated with polyethylene glycol 4000 to obtain TSAb crude extracts acting on FRTL cells, and the Kamin-containing drug serum was divided into low, medium and high concentration groups. In the control group, after co-cultured with FRTL cells, the growth of the cells was detected by MTT assay, and the expression of bcl-2 and bax mRNA was detected by in situ hybridization. Results The TSAb positive serum crude extract promoted the growth of FRTL cells. After co-cultured with different concentrations of Jianing Fang and FRTL cells, the cell growth was inhibited in a concentration-dependent manner, and the effect was significant at high dose (P<0.01); The positive expression rate of bcl-2 was decreased in the group, and the positive expression rate of bax was increased in the middle and high concentrations of Jianing group. The ratio of bcl-2/bax was decreased in Jianing group, and the effect was gradually increased from medium to high concentrations. CONCLUSION: Jia Ning can effectively inhibit the proliferation of FRTL cells induced by TSAb and induce its apoptosis. This may be one of the mechanisms of the treatment of Graves disease.