The molecular mechanism for expression regulation of IL-6 via H3K9 deacetylation in murine macrophage induced by alpinetin

HU Ke; HUANG Minjiang; DUAN Yufeng; QIAN Hong; CAO Xiangyu; LIU Lijing

doi:10.13431/j.cnki.immunol.j.20170083

Abstract：In order to reveal the role of H3K9 deacetylation in IL-6 expression from RAW246.7 caused by alpinetin,we detected the acetylation state of H3K9 and the expression level of transcription factors (P40 and CREB) combined in promoter region or nucleus.Firstly,RAW246.7 were divided into control group,alpinetin groups (with final concentration of 50,100,200 μg/ml),200 μg/ml alpinetin+GW9662 group,200 μg/ml alpinetin+ HDAC1,and Pgene SiRNA group.After incubation,concentration of IL-6 in media was tested by ELISA,while Western blot was used to detect P40,CREB and PPAR in nucleus;content of deacetylated H3K9,P40 and CREB binding in promoter region of IL-6 were tested by chip-qPCR.Date showed that alpinetin promoted the expression of PPAR and deacetylated H3K9 in IL-6 promoter in a dose-dependent manner,meanwhile inhibited generation of IL-6,accompanied by a decline of transcription factor both binding in promoter and nucleus.Compared with 200 μg/ml alpinetin group,GW9662 could suppress the promotion of PPAR and deacetylated H3K9 binding caused by alpinetin,recover the content and binding capacity of transcription factors in nucleus,and improve synthesis of IL-6;HDAC1 intervention had no influence on total synthesis of P40 and CREB in nucleus and promotion of PPAR,but could also repress the promotion of deacetylated H3K9 and promote the down-regulated binding content of P40 and CREB,thus increased synthesis of IL-6,which was still lower than GW9662 group.In conclusion,the PPAR agonist alpinetin could promote deacetylation state of H3K9 combined in the promoter of IL-6 via specific deacetylase,which could affect the binding capacity of corresponding transcription factors,thus regulate gene expression.However,synthesis of transcription factors are not interfered by deacetylation of H3K9.