黄芪甲苷调控p38 MAPK/NF-κB信号通路改善甲亢大鼠细胞凋亡的机制

Mechanism of Astragaloside Regulating p38 MAPK/NF-κB Signaling Pathway to Improve Apoptosis in Hyperthyroidism Rats

目的:通过分析黄芪甲苷干预p38MAPK/NF-κB信号通路,分析其改善甲亢模型大鼠甲状腺组织细胞凋亡的机制。方法:将60只SPF级Wistar大鼠随机分为空白组、模型组、甲巯咪唑组和黄芪甲苷低、高剂量组,每组12只。空白组不做处理,其他组制备甲亢大鼠模型后进行干预治疗。比较5组大鼠干预前后基础代谢情况;放疫法检测血清三碘甲状腺原氨酸(T3)、甲状腺素(T4)和促甲状腺激素(TSH)的浓度;ELISA法检测大鼠血清中IL-1β、IL-6、TNF-α及VEGF含量变化;HE染色观察各组大鼠甲状腺组织病理学改变;TUNEL法检测各组甲状腺组织中的细胞凋亡情况;Western Blot及实时荧光定量PCR法检测各组甲状腺组织中NF-κBp65、p38MAPK蛋白及mRNA表达水平。结果:造模后,与空白组比较,模型组大鼠进食及进水量增加,体质量减少,T3、T4、TSH及IL-1β、IL-6、TNF-α、VEGF含量水平增高(均P<0.05),组织病理损害程度增加,细胞凋亡水平增高,甲状腺组织中NF-κBp65、p38MAPK蛋白及mRNA表达升高(P<0.05)。与模型组比较,甲巯咪唑组及黄芪甲苷干预组大鼠,进食及进水量减少,体质量明显增加,T3、T4、TSH及IL-1β、IL-6、TNF-α及VEGF含量水平降低(均P<0.05),组织病理损害程度明显改善,细胞凋亡水平降低,甲状腺组织中NF-κBp65、p38MAPK蛋白及mRNA表达明显降低(P<0.05),其中黄芪甲苷高剂量组改善程度显著优于黄芪甲苷低剂量组(P<0.01)。结论:黄芪甲苷能够有效改善甲亢大鼠甲状腺组织细胞凋亡,降低炎症因子表达,其机制可能与黄芪甲苷能够调控p38MAPK/NF-κB信号通路表达有关。

Objective:To investigate the effect of Astragaloside on the cellular damage of thyroid tissue in hyperthyroidism model rats by intervening p38MAPK/NF-κBp65 signaling pathway.Methods:A total of 60 SPF grade Wistar rats were randomly divided into blank group,model group,methimazole group and Astragaloside low and high dose groups,with 12 rats in each group.The blank group was left untreated,and the other groups were treated with intervention after preparation of hyperthyroidism rat models.The basal metabolism of the rats in the five groups was compared before and after the intervention.The expression levels of NF-κBp65,p38MAPK protein and mRNA in thyroid tissues were detected by Western Blot and real-time fluorescence quantitative PCR.Results:After modeling,compared with the blank group,rats in the model group had increased food and water intake,decreased body mass,increased levels of T3,T4,TSH and IL-1β,IL-6,TNF-α,VEGF content(all P<0.05),increased histopathological damage,increased levels of apoptosis,and increased NF-κBp65,p38MAPK protein and mRNA expression in thyroid tissue were elevated(P<0.05).Compared with the model group,rats in the methimazole and astragaloside intervention groups showed decreased food and water intake,significantly increased body mass,decreased levels of T3,T4,TSH and IL-1β,IL-6,TNF-αand VEGF(all P<0.05),significantly improved histopathological damage,decreased levels of apoptosis,and increased levels of NF-κBp65,p38MAPK protein and mRNA in thyroid tissues(P<0.05),p38MAPK protein and mRNA expression in thyroid tissues were significantly reduced(P<0.05),in which the improvement in the high-dose group of astragaloside was significantly better than that in the low-dose group of astragaloside(P<0.01).Conclusion:Astragaloside can effectively improve the apoptosis and reduce the inflammatory factor expression in the thyroid tissue,which may be related to the regulation of the p38MAPK/NF-κB signaling pathway.