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目的:观察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)甲状腺球蛋白基因(TGmRNA)、甲状腺过氧化物酶基因(TPOmRNA)表达的影响,探讨该方治疗格雷夫斯病(Graves)的可能机理。方法:用Graves病患者血清经聚乙二醇4000处理获得的TSAb粗提物,用MTT法及逆转录聚合酶链反应(RT-PCR)观察甲宁方对TSAb作用下FRTL细胞生长和TGmRNA、TPOmRNA表达的影响。结果:TSAb阳性血清粗提物促进FRTL细胞生长,模型组与正常组比较,差异有显著性意义(P<0.05)。甲宁方各浓度组对TSAb阳性血清粗提物刺激下的FRTL细胞生长均有抑制作用,并随甲宁方浓度增加抑制加强,其中高浓度甲宁方组与模型组比较,差异有显著性意义(P<0.05)。甲亢灵片也可抑制TSAb阳性血清粗提物刺激下的FRTL细胞生长,但与模型组比较,差异无显著性意义(P>0.05)。模型组FRTL细胞在TSAb阳性血清粗提物作用下,TGmRNA、TPOmRNA表达均较正常组增加,TGmRNA约增加1.8倍,TPOmRNA约增加1.7倍,提示TSAb可上调FRTL细胞TGmRNA、TPOmRNA表达。中、高浓度甲宁方组FRTL细胞TGmRNA表达较模型组低,高浓度甲宁方组可明显抑制FRTL细胞TPOmRNA表达的上调作用,说明甲宁方可抑制TSAb诱导的FRTL细胞TGmRNA、TPOmRNA表达。甲亢灵片也可抑制TSAb诱导的FRTL细胞TGmRNA、TPOmRNA表达,但作用不及中、高浓度甲宁方。结论:甲宁方可抑制Graves病的自身抗体TSAb对甲状腺细胞生长的刺激,推测抑制TGmRNA、TPOmRNA可能是甲宁方临床降低三碘甲状腺原氨酸(T_3)、甲状腺素(T_4)的主要机理之一。
Objective: To observe the effect of Jianing prescription on the expression of thyroid globulin gene (TGmRNA) and thyroid peroxidase gene (TPOmRNA) in Fisher rat thyroid cell line (FRTL) under the action of thyroid stimulating antibody (TSAb). The possible mechanism of Graves disease. METHODS: The TSAb crude extracts obtained from the serum of patients with Graves disease treated with polyethylene glycol 4000 were used to observe the growth and TG mRNA of FRTL cells under the action of TSNA and Methyltransferase (MTT) and reverse transcription polymerase chain reaction (RT-PCR). The effect of TPO mRNA expression. Results: TSAb positive serum crude extract promoted the growth of FRTL cells, and there was significant difference between the model group and the normal group (P<0.05). Each concentration group of Jia Ning Fang inhibited the growth of FRTL cells stimulated by crude extracts of TSAb positive serum and inhibited the growth of FRTL cells with the increase of the concentration of Jia Ning prescription. Compared with the model group, the high concentration of Ning Fang prescription group had significant difference. Significance (P < 0.05). Hyperthyroidism tablets also inhibited the growth of FRTL cells stimulated with TSAb-positive serum extracts, but there was no significant difference compared with the model group (P>0.05). The expression of TG mRNA and TPO mRNA in the FRTL cells of the model group was higher than that in the normal group. The expression of TG mRNA and TPO mRNA was increased by 1.8 times and TPO mRNA was increased by 1.7-fold, suggesting that TSAb can upregulate the expression of TG mRNA and TPO mRNA in FRTL cells. The expression of TGmRNA in FRTL cells was lower than that in the model group in the high-concentration and high-concentration Jianing formula group. The high-concentration Jianing formula group could significantly inhibit the up-regulation of TPOmRNA expression in FRTL cells, suggesting that MNG could inhibit the expression of TGmRNA and TPOmRNA in FRTL cells induced by TSAb. Hyperthyroidism tablets also inhibited TSAb-induced TG mRNA and TPO mRNA expression in FRTL cells, but the effect was lower than that of high-concentration formazan. Conclusion: MJN can inhibit the stimulation of thyroid cell growth by autoantibodies TSAb of Graves’ disease. It is speculated that inhibiting TG mRNA and TPO mRNA may be the main mechanism of the clinical reduction of triiodothyronine (T_3) and thyroxine (T_4) in MKN. one.