基于氧化应激及肠道菌群探讨逍遥补肾方对实验性自身免疫性甲状腺炎模型大鼠作用机制的影响  

Effect of Xiaoyao Bushen Formula(逍遥补肾方)on Mechanism of Experimental Autoimmune Thyroiditis Model Rats Based on Oxidative Stress and Intestinal Flora

作　　者：刘婧 魏东升 安杨 孙欣[3] 张国栋 伊桐凝[4] 张兰[4] LIU Jing;WEI Dongsheng;AN Yang;SUN Xin;ZHANG Guodong;YI Tongning;ZHANG Lan(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Key Laboratory of Ministry of Education for TCM Viscera-State Theory and Applications,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;Shenyang Second Hospital of Traditional Chinese Medicine,Shenyang 110101,Liaoning,China;Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,Liaoning,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁中医药大学中医脏象理论及应用教育部重点实验室,辽宁沈阳110847 [3]沈阳市第二中医医院,辽宁沈阳110101 [4]辽宁中医药大学附属医院,辽宁沈阳110032

出　　处：《中华中医药学刊》2024年第10期64-69,I0011-I0014,共10页Chinese Archives of Traditional Chinese Medicine

基　　金：国家中医药管理局全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号)。

摘　　要：目的通过观察逍遥补肾方对实验性自身免疫性甲状腺炎模型(experimental autoimmune thyroiditis,EAT)大鼠氧化应激及肠道菌群的影响,探讨逍遥补肾方的疗效机制。方法将65只大鼠随机分为正常组10只,造模组55只。造模组采用高碘水喂养联合甲状腺球蛋白皮下注射复刻EAT大鼠模型,造模成功的EAT大鼠随机分为模型组、硒酵母组及逍遥补肾组,每组17只。硒酵母组予25.2 mg/(kg·d)硒酵母片水溶液,逍遥补肾组予11.79 g/(kg·d)逍遥补肾方溶液,正常组、模型组灌胃给予等体积的蒸馏水。连续灌胃8周后,采用酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法检测各组大鼠血清游离三碘甲状腺原氨酸(free triiodothyronine3,FT3)、游离甲状腺素(free thyroxine,FT4)、促甲状腺激素(thyroid stimulating hormone,TSH)、甲状腺球蛋白抗体(thyroglobulin antibody,TGAb)、甲状腺过氧化物酶抗体(thyroid peroxidase antibody,TPOAb)、超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malonic dialdehyde,MDA)、谷胱甘肽过氧化物酶(glutathione peroxidase,GPx)水平,采用免疫荧光检测活性氧(reactive oxygen species,ROS)水平。收集各组大鼠的粪便进行16S rDNA高通量测序,分析肠道菌群多样性和物种丰度的变化,通过相关性分析筛选逍遥补肾方改善EAT大鼠的相关菌群。结果与正常组相比,模型组大鼠血清TSH、SOD、SOD/MDA、GPx明显下降,TPOAb、TGAb、MDA水平明显升高,ROS的免疫荧光强度明显增强(P<0.01)。给予治疗药物后,逍遥补肾组及硒酵母组的上述指标均较模型组有一定改善(P<0.01或P<0.05)。16S rDNA测序结果显示,逍遥补肾方能够调节厚壁菌门、拟杆菌门等相对丰度调节EAT大鼠的肠道菌群紊乱。脱硫弧菌属及粪球菌属是逍遥补肾组的优势菌群。结论逍遥补肾方能够调节EAT大鼠血清TSH及甲状腺双抗体水平,改善氧化应激状态,其作用机制可能与调节多种肠道菌�Objective To explore the therapeutic mechanism of Xiaoyao Bushen Formula(逍遥补肾方)by observing the effect of it on the oxidative stress and intestinal flora in experimental autoimmune thyroiditis(EAT)rats.Methods Sixty-five rats were randomly divided into the control group(10 rats)and the EAT model group(55 rats).The EAT model group used high iodine water feeding combined with subcutaneous injection of thyroglobulin to replicate the EAT model.The successfully modeled EAT rats were randomly divided into the model group,the selenium yeast(Se)group and Xiaoyao Bushen Formula group,with 17 rats in each group.The Se group was given 25.2 mg/(kg·d)selenium yeast tablet aqueous solution,the Xiaoyao Bushen Formula group was given 11.79 g/(kg·d)Xiaoyao Bushen Formula and the control group and the model group were given the equal volume of distilled water by gavage.After 8 weeks of continuous intragastric administration,the levels of serum free triiodothyronine 3(FT3),free thyroxine(FT4),thyroid stimulating hormone(TSH),thyroglobulin antibody(TGAb),thyroid peroxidase antibody(TPOAb),superoxide dismutase(SOD),malonic dialdehyde(MDA)and glutathione peroxidase(GPx)of rats in each group were detected by enzyme-linked immunosorbent assay(ELISA),and the level of reactive oxygen species(ROS)was detected by immunofluorescence.The feces of rats in each group were collected for 16S rDNA high-throughput sequencing,the changes of intestinal flora diversity and species abundance were analyzed,and the relevant flora of Xiaoyao Bushen Formula to improve EAT rats was screened through correlation analysis.Results Compared with those of the control group,the levels of serum TSH,SOD,SOD/MDA and GPx of the model group rats were significantly reduced,while the levels of TPOAb,TGAb and MDA were markedly increased,and the immune fluorescence intensity of ROS was significantly enhanced.After administering treatment drugs,the above indicators in the Xiaoyao Bushen Formula group and Se group improved compared to those of the model group(P<0

关 键 词：实验性自身免疫性甲状腺炎 逍遥补肾方 氧化应激 肠道菌群 

分 类 号：R289.5[医药卫生—方剂学]