补中益气汤调控TLR4/NF-κB/AIM2信号通路改善自身免疫性甲状腺炎小鼠甲状腺炎症损伤的作用机制  

Mechanism of Buzhong Yiqitang in Ameliorating Thyroiditis Damage in AIT Mice by Modulating TLR4/NF-κB/AIM2 Signaling Pathway

作　　者：赵卓 金哲[2] 李铮铮 郭宣麟 李佳韵 高彤然 李品[3] 王智民[3] 尹远平[2] 刘子玉 杨潇[2] ZHAO Zhuo;JIN Zhe;LI Zhengzheng;GUO Xuanlin;LI Jiayun;GAO Tongran;LI Pin;WANG Zhimin;YIN Yuanping;LIU Ziyu;YANG Xiao(Liaoning University of Traditional Chinese Medicine(TCM),Shenyang 110847,China;The Second Affiliated Hospital of Liaoning University of TCM,Shenyang 110034,China;Affiliated Hospital of Liaoning University of TCM,Shenyang 110032,China;Liaoning Institute of TCM,Shenyang 110034,China)

机构地区：[1]辽宁中医药大学,沈阳110847 [2]辽宁中医药大学附属第二医院,沈阳110034 [3]辽宁中医药大学附属医院,沈阳110032 [4]辽宁省中医药研究院,沈阳110034

出　　处：《中国实验方剂学杂志》2024年第18期1-9,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(82274455,82104805);辽宁省教育厅重点攻关项目(JYTZD2023196);辽宁省科技厅联合基金项目(2023-MSLH-154);尹远平全国名老中医药专家传承工作室项目(202205)。

摘　　要：目的:基于Toll样受体4(TLR4)/核转录因子-κB(NF-κB)/黑色素瘤缺乏因子2(AIM2)炎性小体信号通路探索补中益气汤改善自身免疫性甲状腺炎(AIT)小鼠炎症损伤的作用机制。方法:选用基因易感8周龄NOD.H-2h4小鼠120只,随机分为空白组、模型组、补中益气汤低、中、高剂量组(4.78、9.56、19.12 g·kg^(-1))、西药组(硒酵母片,3.033×10^(-5) g·kg^(-1))。各组AIT模型小鼠自由饮用0.05%碘化钠水溶液8周,建立AIT模型,空白组自由饮用蒸馏水。依据分组灌胃各给药组小鼠药液8周后取材,肉眼观察甲状腺组织肿胀情况,称量脾脏质量,酶联免疫吸附测定法(ELISA)检测血清炎症因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量,实时荧光定量聚合酶链式反应(Real-time PCR)检测甲状腺组织高迁移率族蛋白1(HMGB1)、TLR4、AIM2、NF-κB p65、凋亡相关斑点样蛋白(ASC)、胱天蛋白酶-1(Caspase-1)、IL-1β mRNA的表达,蛋白免疫印迹法(Western blot)检测甲状腺组织HMGB1、TLR4、AIM2、NF-κB p65、磷酸化(p)-NF-κB p65、ASC、Caspase-1、IL-1β蛋白的表达,免疫荧光染色观察小鼠甲状腺组织HMGB1、AIM2、NF-κB p65的蛋白表达情况。结果:与空白组比较,模型组小鼠甲状腺组织明显肿胀,脾脏质量明显增加,甲状腺组织HMGB1、TLR4、NF-κB p65、AIM2、ASC、Caspase-1、IL-1β表达显著升高(P<0.01);与模型组比较,补中益气汤各剂量组小鼠甲状腺组织肿胀情况好转,脾脏质量明显下降,甲状腺组织HMGB1、TLR4、AIM2、NF-κB p65、p-NF-κB p65、ASC、Caspase-1、IL-1β表达明显降低(P<0.05,P<0.01)。结论:补中益气汤可有效改善AIT的炎症损伤,调控TLR4/NF-κB/AIM2炎性小体信号通路的异常活化可能是其干预机制之一。Objective To explore the mechanism of Buzhong Yiqitang in ameliorating inflammatory injury in autoimmune thyroiditis(AIT)mice based on the Toll-like receptor 4(TLR4)/nuclear transcription factor-kappa B(NF-κB)/absent in melanoma 2(AIM2)inflammasome signaling pathway.Method The 120 genetically susceptible 8-week-old NOD.H-2h4 mice were selected and randomly divided into control group,model group,low,medium and high dose groups of Buzhong Yiqitang(4.78,9.56,19.12 g·kg^(-1)),and western medicine group(selenium yeast tablets,3.033×10^(-5) g·kg^(-1)).The AIT model mice in each group drank ad libitum 0.05%sodium iodide aqueous solution for 8 weeks to establish the AIT model,and the control group drank ad libitum distilled water.Eight weeks later,the mice in each dosing group were divided into groups and gavage.The swelling of thyroid tissue was observed with the naked eye,and the weight of spleen was weighed.The content of serum inflammatory factors interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)was measured by enzyme-linked immunosorbent assay(ELISA),and Real-time PCR was used to detect the expression of HMGB1,TLR4,AIM2,NF-κB p65,apoptosis-associated speck-like protein(ASC),cysteinyl aspartate-specific protease-1(Caspase-1),IL-1βmRNA.Western blot was used to detect the expression of high motility group protein 1(HMGB1),TLR4,AIM2,NF-κB p65,phosphorylation(p)-NF-κB p65,ASC,Caspase-1,and IL-1βproteins in thyroid tissue,and immunofluorescence staining was used to observe the protein expression of HMGB1,AIM2,and NF-κB p65 in thyroid tissue of mice.Result Compared with the control group,the thyroid tissue of mice in the model group was significantly swollen,the spleen quality was significantly increased,and the expression of HMGB1,TLR4,NF-κB p65,AIM2,ASC,Caspase-1,IL-1βin thyroid tissue was significantly increased(P<0.01).Compared with the model group,the swelling of thyroid tissue in mice in each dose group of Buzhong Yiqitang was improved,the quality of spleen was significantly red

关 键 词：补中益气汤 Toll样受体4(TLR4)/核转录因子-κB(NF-κB)信号通路 黑色素瘤缺乏因子2(AIM2)炎性小体 自身免疫性甲状腺炎 炎症 

分 类 号：R2-0[医药卫生—中医学] R33R289R593R24