Potential Mechanism of Buzhong Yiqitang for Autoimmune Thyroiditis Based on miRNA Sequencing / 中国实验方剂学杂志

ABSTRACT

ObjectiveTo determine the influence of Buzhong Yiqitang on miRNA expression in thyroid tissues of mice with autoimmune thyroiditis （AIT）. MethodThirty female 8-week-old NOD.H-2h4 mice 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. ResultCompared 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-6 and 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. ConclusionBushong Yiqitang might treat autoimmune thyroiditis by regulating 6 miRNAs.