Zhao Zhuo, Liu Ziyu, Song Nan, Cao Huimin, Chen Yiran, Chen Si, Wang Zhimin, Jin Zhe, Yang Xiao
The second Clinical College of Liaoning University of Traditional Chinese Medicine, Shenyang, Liaoning 110847, China.
Liaoning Institute of Traditional Chinese Medicine, Shenyang, Liaoning 110034, China.
Comb Chem High Throughput Screen. 2024 Aug 12. doi: 10.2174/0113862073294048240801113424.
Autoimmune thyroiditis (AIT) is the most common autoimmune thyroid disease. In recent decades, its incidence and prevalence have sharply increased. Yiqi Huatan Huoxue recipe is a traditional Chinese medicine formula we use to treat AIT. Its clinical efficacy is clear, but the specific mechanism remains unclear. This study aims to explore whether pyroptosis mediated by the SIRT1/NF-κB/NLRP3 signaling pathway is one of the therapeutic mechanisms of Yiqi Huatan Huoxue recipe.
Forty 8-week-old female NOD.H-2h4 mice were randomly divided into four groups: the normal group (NG), model group (MG), Yiqi Huatan Huoxue recipe group (YG), and western medicine group (selenium yeast tablet, SeG). The normal group was gavaged with distilled water, while the remaining groups were gavaged with 0.05% sodium iodide (NaI) solution for 8 weeks. After the AIT animal model formed naturally, the mice were euthanized by gavage after 8 weeks. Hematoxylin-eosin staining was used to observe thyroid tissue changes, and enzymelinked immunosorbent assay (ELISA) was used to detect serum anti-thyroglobulin antibodies (TGAb) and mouse anti-thyroid peroxidase antibodies (TPOAb). Real-time quantitative PCR (qRT-PCR), Western blot, and immunohistochemistry were used to detect the expression of sirtuin 1 (SIRT1), nuclear factor κB p65 (NF-κB p65), nod-like receptor protein 3 (NLRP3), apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), caspase- 1, gasdermin D (GSDMD), and interleukin (IL)-1β in thyroid tissue.
Compared with the NG group, the thyroid structure of rats in the MG group was severely damaged, with significant lymphocyte infiltration, significantly increased serum TGAb and TPOAb levels, and significantly increased expression levels of SRIT1, NF-κB p65, NLRP3, ASC, Caspase-1, GSDMD, IL-1β mRNA, and protein. Compared with the MG group, the thyroid structure damage and lymphocyte infiltration in rats of each treatment group were improved, and the serum TGAb, TPOAb, SRIT1, NF-κB p65, NLRP3, ASC, Caspase-1, GSDMD, IL-1β mRNA, and protein expression levels were significantly reduced.
Yiqi Huatan Huoxue recipe can alleviate thyroid structural damage in AIT mice, and its mechanism may be related to the upregulation of SIRT1, NF-κB deacetylation, and inhibition of NLRP3-mediated pyroptosis.
自身免疫性甲状腺炎（AIT）是最常见的自身免疫性甲状腺疾病。近几十年来，其发病率和患病率急剧上升。益气化痰活血方是我们用于治疗AIT的中药方剂。其临床疗效明确，但具体机制尚不清楚。本研究旨在探讨SIRT1/NF-κB/NLRP3信号通路介导的细胞焦亡是否为益气化痰活血方的治疗机制之一。
将40只8周龄雌性NOD.H-2h4小鼠随机分为四组：正常组（NG）、模型组（MG）、益气化痰活血方组（YG）和西药组（硒酵母片，SeG）。正常组灌胃蒸馏水，其余各组灌胃0.05%碘化钠（NaI）溶液8周。自然形成AIT动物模型后，8周后通过灌胃处死小鼠。采用苏木精-伊红染色观察甲状腺组织变化，采用酶联免疫吸附测定（ELISA）检测血清抗甲状腺球蛋白抗体（TGAb）和小鼠抗甲状腺过氧化物酶抗体（TPOAb）。采用实时定量PCR（qRT-PCR）、蛋白质印迹法和免疫组织化学法检测甲状腺组织中沉默调节蛋白1（SIRT1）、核因子κB p65（NF-κB p65）、NOD样受体蛋白3（NLRP3）、含半胱天冬酶募集结构域的凋亡相关斑点样蛋白（ASC）、半胱天冬酶-1、gasdermin D（GSDMD）和白细胞介素（IL）-1β的表达。
与NG组相比，MG组大鼠甲状腺结构严重受损，淋巴细胞浸润明显，血清TGAb和TPOAb水平显著升高，SRIT1、NF-κB p65、NLRP3、ASC、半胱天冬酶-1、GSDMD、IL-1β mRNA和蛋白表达水平显著升高。与MG组相比，各治疗组大鼠甲状腺结构损伤和淋巴细胞浸润均有所改善，血清TGAb、TPOAb、SRIT1、NF-κB p65.NLRP3、ASC、半胱天冬酶-1、GSDMD、IL-1β mRNA和蛋白表达水平显著降低。
益气化痰活血方可减轻AIT小鼠甲状腺结构损伤，其机制可能与上调SIRT1、NF-κB去乙酰化及抑制NLRP3介导的细胞焦亡有关。
