摘要
目的:探讨补中益气汤对自身免疫性甲状腺炎(AIT)小鼠甲状腺组织miRNA表达的影响。方法:选择NOD.H-2h4小鼠共30只,随机分为正常组、模型组、补中益气汤组(BG组),每组各10只。根据分组给予小鼠含0.05%碘化钠水8周构建小鼠AIT模型后,灌胃给药8周取材。观察每组小鼠甲状腺组织病理改变情况,并对每组小鼠甲状腺组织差异miRNA进行实验验证及生物信息学分析。结果:与正常组比较,模型组甲状腺组织炎性细胞浸润明显,血清甲状腺球蛋白抗体(TgAb)水平显著升高(P<0.01);与模型组比较,BG组甲状腺组织炎症程度减轻,血清TgAb水平显著降低(P<0.01)。与正常组比较,模型组小鼠白细胞介素(IL)-6、IL-17表达显著增高(P<0.01),IL-1β表达明显降低(P<0.05);与模型组比较,BG组IL-1β、IL-6、IL-17表达明显降低(P<0.05)。与正常组比较,模型组得到154个差异表达miRNA;与模型组比较,BG组得到112个差异表达miRNA。实时荧光定量聚合酶链式反应(Real-time PCR)结果表明miR-326-3p、miR-128-3p、miR-223-5p、miR-141-3p、miR-871-3p、miR-204-3p表达与测序结果趋势一致。基因本体(GO)功能富集于T细胞活化调节、氧化应激、miRNA结合等方面;京都基因与基因组百科全书(KEGG)通路富集于磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路、丝裂原活化蛋白激酶(MAPK)信号通路、环磷酸腺苷(cAMP)信号通路等途径。差异miRNA预测得到3个关键基因,果蝇母源抗皮肤生长因子蛋白(Smad3)、Janus相关激酶2(JAK2)、信号转导和转录激活因子3(STAT3)。结论:补中益气汤可能通过调节6种miRNA干预自身免疫性甲状腺炎。
Objective:To determine the influence of Buzhong Yiqitang on miRNA expression in thyroid tissues of mice with autoimmune thyroiditis(AIT).Method:Thirty female 8-week-old NOD.H-2h4mice were randomly assigned into normal control group,model group,and Buzhong Yiqitang group(BG),10 in each group.Mice were subjected to a diet containing 0.05% sodium iodide for 8 weeks to build the AIT mouse model.After 8 weeks of administration(ig),samples were collected.A thyroid biopsy was performed on each group of mice,and differential miRNAs in thyroid tissues from each group of mice were analyzed based on experimental validation and bioinformatics.Result:Compared with the conditions of normal control group,thyroid lymphocytes had significant inflammatory infiltration,and there was an increase in serum TgAb level and interleukin(IL)-6 and IL-17 expression and a decrease in IL-1β expression in mice of the model group(P<0.05,P<0.01).In addition,154 differentially expressed miRNAs were found.Compared with the conditions of model group,the degree of thyroid tissue inflammation was alleviated,and serum TgAb level,and IL-1β,IL-6and IL-17 expression of mice treated with the Buzhong Yiqitang were reduced(P<0.05,P<0.01).Additionally,112 differentially expressed miRNAs were identified in the BG group.Validation using real-time polymerase chain reaction(Real-time PCR)showed the same trend for miR-326-3p,miR-128-3p,miR-223-5p,miR-141-3p,miR-871-3p,and miR-204-3p as that obtained from miRNA sequencing.In particular,gene ontology(GO)functions were enriched for regulation of T cell activation,oxidative stress,and miRNA binding.Pathways identified by Kyoto encyclopedia of genes and genomes(KEGG)database tended to be enriched in phosphatidylinositol 3-kinases(PI3K)/protein kinase B(Akt),mitogen-activated protein kinase(MAPK),and cyclic adenosine monophosphate(cAMP)signaling pathways.Based on miRNA prediction differences,three key genes were identified:SMAD3,JAK2,and STAT3.Conclusion:Bushong Yiqitang might treat autoimmune thyroiditis by regulating 6 miRNAs.
作者
刘子玉
王智民
宋囡
曹慧敏
陈丝
尹远平
高天舒
杨潇
LIU Ziyu;WANG Zhimin;SONG Nan;CAO Huimin;CHEN Si;YIN Yuanping;GAO Tianshu;YANG Xiao(Graduate School of Liaoning University of Traditional Chinese Medicine(TCM),Shenyang 110847,China;Affiliated Hospital of Liaoning University of TCM,Shenyang 110032,China;Innovative Engineering Technology Center of TCM,Liaoning University of TCM,Shenyang 110847,China;The Second Affiliated Hospital of Liaoning University of TCM,Shenyang 110034,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第21期192-200,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
辽宁中医药大学中医脏象理论及应用国家教育部重点实验室开放基金项目(zyzx1804)
辽宁省科技厅自然基金项目(2019-MS-207)
国家自然科学基金青年基金项目(81503474,82104805)
国家自然科学基金面上项目(81874441)
辽宁省教育厅科学技术研究项目(L201923)
沈阳市中青年科技创新人才支持计划项目(RC200370)
尹远平全国名老中医药专家传承工作室。