亚甲方下调miR-155改善炎症反应治疗亚急性甲状腺炎的机制研究OA

Objective To investigate the role and mechanism of Yajia Formula in improving the progress of subacute thyroiditis(SAT)inflammatory response.Methods Drug-containing serum of Yajia Formula was prepared.Cells were transfected with oligoRNA(miR-155 mimics,miR-155 inhibitor,mimics NC,and inhibitor NC)using Lipofectamine 2000 to regulate miR-155 expression.After pretreatment with a high dose of Yajia Formula for 1 hour,the cells were stimulated with TNF-α(25 ng/mL).qRT-PCR was performed to detect the mRNA expression of miR-155,SOCS1,JAK2,and STAT3.Western blot was employed to detect the protein levels of SOCS1,JAK2,STAT3,p-JAK2,and p-STAT3.Results ①Compared with TNF-α group,low-dose Yajia Formula could reduce the release of IL-6,lower the mRNA expression of miR-155,and decrease the total protein levels of JAK2 and STAT3,and phosphorylation level of STAT3(P<0.01,P<0.05).Medium-dose Yajia Formula could reduce the release of the protein expression of IL-1β and IL-8(P<0.05).High-dose Yajia Formula could downregulate phosphorylation level of JAK2(P<0.05).② Compared with miR-155 overexpression group,Yajia Formula could reverse the increase in miR-155(P<0.01),downregulate the phosphorylation levels of JAK2 and STAT3(P<0.01),and upregulate the mRNA expression of SOCS1 and protein levels(P<0.01,P<0.05);Compare with miR-155 inhibition group,Yajia Formula could further downregulate miR-155 expression(P<0.05),reduce the total protein expression and phosphorylation levels of JAK2 and STAT3(P<0.01,P<0.05),and upregulate SOCS1 mRNA expression(P<0.01).Conclusion Yajia Formula can inhibit the inflammatory response of cells by downregulating miR-155,which leads to increased expression of the target protein SOCS1 and suppression of JAK2/STAT3 signaling pathway activation.