北京中医药大学学报2025，Vol.48Issue(4)：529-541,13.DOI:10.3969/j.issn.1006-2157.2025.04.010
补中益气汤调控自身免疫性甲状腺炎小鼠巨噬细胞极化的机制研究
Study on the mechanism of Buzhong Yiqi Decoction regulating macrophage polarization in mice with autoimmune thyroiditis
摘要
Abstract
Objective To explore the mechanism of Buzhong Yiqi Decoction in modulating macrophage polarization and intervening in autoimmune thyroiditis(AIT)mice.Methods Using the random number table method,48 SPF-grade NOD.H-2h4 mice were assigned to the normal,model,low-dose(4.10 g/kg),medium-dose(8.19 g/kg),high-dose group(16.38 g/kg)of Buzhong Yiqi Decoction,and selenium yeast tablet(0.026 mg/kg)groups,with eight mice in each group.All groups,except the normal group,were free to drink high iodine water(0.05%sodium iodide)to prepare AIT mouse models for 8 consecutive weeks.After the modeling was complete,each treatment group was orally administered with the corresponding medication,while the normal and model groups were orally administered with an equal volume of distilled water once a day for 8 consecutive weeks.High-performance liquid chromatography with an oscillometric refractive detector was used to analyze the content of Astragaloside Ⅳ in Buzhong Yiqi Decoction.Hematoxylin and eosin staining was used to observe the pathological morphology of mouse thyroid tissue.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of serum thyroid peroxidase antibody(TPO-Ab),thyroglobulin antibody(TgAb),interleukin(IL)-6,IL-10,and tumor necrosis factor-α(TNF-α).An immunofluorescence assay was used to detect the positive area percentage of M1 and M2 macrophages in mouse thyroid tissue.Flow cytometry assay was used to detect macrophage polarization in mouse spleen tissue.Real-time fluorescence quantitative PCR was used to detect the mRNA expression of nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3(NLRP3),nuclear factor kappa B inhibitory protein α(IκBα),and nuclear factor-κB(NF-κB)p65 in mouse spleen tissue.Western blotting was used to detect the expression of the phosphorylated IκBα(p-IκBα),phosphorylated NF-κB p65(p-NF-κB p65),and NLRP3 protein in mouse spleen tissue.Results The content of Astragaloside Ⅳ in Buzhong Yiqi Decoction was(7.09±0.06)g/L.Compared to the normal group,significant lymphocyte infiltration was observed in the thyroid tissue of mice in the model group.The levels of serum TPO-Ab,TgAb,IL-6,and TNF-α increased(P<0.05).The positive area percentage of M1 macrophages in thyroid tissue increased(P<0.05).The proportion of M1 macrophages and M1/M2 in spleen tissue increased(P<0.05).The relative expression levels of NF-κB p65 and NLRP3 mRNA in spleen tissue increased(P<0.05).The relative expression of p-IκBα,p-NF-κB p65,and NLRP3 proteins increased(P<0.05).Compared to the model group,the inflammation infiltration degree in the thyroid tissue of mice in each dose group of Buzhong Yiqi Decoction and selenium yeast tablet group was reduced,the serum TPO-Ab,TgAb,IL-6,TNF-α content was decreased,the spleen tissue M1/M2 was reduced,the expression of NF-κB p65 mRNA was reduced,and the relative expression levels of p-IκBα,p-NF-κB p65 protein were reduced(P<0.05).The Buzhong Yiqi Decoction high-dose and selenium yeast tablets groups showed an increase in IL-10 content,an increase in positive area percentage of M2 macrophages in thyroid tissue,an increase in M2 macrophages proportion in spleen tissue,and a decrease in NLRP3 mRNA and protein relative expression levels(P<0.05).Conclusion Buzhong Yiqi Decoction may regulate macrophage polarization by inhibiting the NF-κB/NLRP3 signaling pathway,thus improving the inflammatory damage in mice with AIT.关键词
补中益气汤/自身免疫性甲状腺炎/核转录因子-κB/核苷酸结合结构域富含亮氨酸重复序列和含热蛋白结构域受体3 信号通路/巨噬细胞极化/小鼠Key words
Buzhong Yiqi Decoction/autoimmune thyroiditis/nuclear factor-κB(NF-κB)/nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3(NLRP3)signaling pathway/macrophage polarization/mice分类
中医学引用本文复制引用
王兰亭,翟钊晗,鞠首信,孔亮,丁杰,肖瑶,陈怡然,王智民..补中益气汤调控自身免疫性甲状腺炎小鼠巨噬细胞极化的机制研究[J].北京中医药大学学报,2025,48(4):529-541,13.基金项目
国家自然科学基金青年科学基金项目(No.82104805) （No.82104805）
辽宁省科技计划联合计划项目(No.2023-MSLH-177,No.2023-MSLH-197) （No.2023-MSLH-177,No.2023-MSLH-197）
辽宁省教育厅面上项目(No.LJKZ0881) （No.LJKZ0881）
辽宁省教育厅高校基本科研项目储备项目(No.2024-JYTCB-067) National Natural Science Foundation of China(No.82104805) （No.2024-JYTCB-067）
