摘要
目的:基于沉默信息调节因子1(SIRT1)/核苷酸结合寡聚结构域样受体蛋白3(NLRP3)信号通路探讨益气化痰活血方通过调控细胞焦亡对自身免疫性甲状腺炎(AIT)小鼠炎症损伤的影响.方法:将40只NOD.H-2h4小鼠分为对照(NG)组、模型(MG)组、益气化痰活血方(YG)组及西药硒酵母(SeG)组,每组10只.NG组饲蒸馏水,其余各组饲0.05％碘化钠水8周后,依据分组灌胃给药8周.HE染色观察甲状腺病理改变;ELISA法检测血清抗甲状腺球蛋白抗体(TGAb)、抗甲状腺过氧化物酶抗体(TPOAb)含量;Real-time PCR和Western Blot分别检测甲状腺SIRT1、核因子κB p65(NF-κB p65)、NLRP3、凋亡相关斑点样蛋白(ASC)、胱天蛋白酶-1(Caspase-1)、消皮素D(GSDMD)、白细胞介素(IL)-1β mRNA及蛋白水平.结果:与NG组比较,MG组小鼠甲状腺淋巴细胞明显浸润,血清TGAb、TPOAb含量及甲状腺组织NF-κB p65、NLRP3、ASC、Caspase-1、GSDMD、IL-1 β mRNA及蛋白表达水平显著升高(P＜0.01,P＜0.05),SIRT1 mRNA及蛋白表达水平显著降低(P＜0.01);与MG组比较,各给药组小鼠甲状腺病理损伤程度有所改善,血清TGAb、TPOAb含量显著降低(P＜0.01,P＜0.05),甲状腺组织NF-κB p65、NLRP3、ASC、Caspase-1、GSDMD、IL-1β mRNA及蛋白表达水平显著降低(除SeG组ASC蛋白外)(P＜0.01),SIRT1 mRNA及蛋白表达水平显著升高(P＜0.01,P＜0.05).结论:益气化痰活血方可缓解AIT小鼠甲状腺炎症损伤,其机制可能与上调SIRT1,去乙酰化NF-κB,进而抑制了NLRP3介导的细胞焦亡有关.
Abstract
Objective:To explore the effects of Yiqi Huatan Huoxue Formula on inflammatory injury in mice with autoimmune thyroiditis(AIT)by regulating cell coke death based on SIRT1/NLRP3 signal pathway.Methods:A total of 40 NOD.H-2h4 mice were divided into control group(NG),model group(MG),Yiqi Huatan Huaxue Formula group(YG)and western medicine Selenium yeast group(SeG),10 mice in each group.The NG group was fed with distilled water,and the other groups were fed with 0.05％sodium iodide water for 8 weeks.The pathological changes of thyroid were observed by HE staining,the contents of TGAb and TPOAb in serum were detected by ELISA,and the levels SIRT1,NF-κB p65,NLRP3,ASC,Caspase-1,GSDMD and IL-1β mRNA and protein levels were detected by Real-time PCR and Western Blot.Results:Compared with the NG group,thyroid lymphocytes in the MG group were significantly infiltrated,serum TGAb and TPOAb levels as well as thyroid tissue mRNA and protein expression levels of NF-κB p65,NLRP3,ASC,Caspase-1,GSDMD,and IL-1β were significantly increased(P＜0.01,P＜0.05),while SIRT1 mRNA and protein expression levels were significantly decreased(P＜0.01).Compared with the MG group,the degree of thyroid pathological damage in mice in each treatment group improved,serum TGAb and TPOAb levels were significantly reduced(P＜0.01,P＜0.05),the mRNA and protein expression levels of NF-κB p65,NLRP3,ASC,Caspase-1,GSDMD,and IL-1 β in thyroid tissue were significantly reduced(except for ASC protein in the SeG group)(P＜0.01),and the expression levels of SIRT1 mRNA and protein were significantly increased(P＜0.01,P＜0.05).Conclusion:Yiqi Huatan Huoxue Formula can alleviate the thyroid structural damage in AIT mice,and its mechanism may be related to the up-regulation of SIRT1,deacetylation of NF-κB,and inhibition of NLRP3-induced cell coke death.
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