摘要
目的探讨BRG1小分子抑制剂BBAi-1在Th17细胞中是否会影响STAT3介导的增强子RORCE2染色质开放。方法检测WT和STAT3结合位点(STAT3-bindingsite,STAT3-BS)缺失小鼠脾脏CD4^(+)T细胞中RORγt基因和蛋白的表达水平以及Th17细胞频率。体外极化的Th17细胞分为WT对照组和BRG1小分子抑制剂(BBAi-1)处理组,检测RORCE2的染色质可及性、RORγt和IL-17A mRNA的表达水平,以及Th17细胞频率。建立EAE小鼠模型,分为WT对照组和BBAi-1给药组,免疫后进行临床评分,第30天处死小鼠,制备脊髓组织石蜡切片进行HE染色和LFB染色,制备脊髓浸润单核细胞悬液进行流式细胞术。结果STAT3-BS的缺失使脾脏CD4^(+)T细胞中RORγt表达和Th17细胞频率减少;BBAi-1使RORCE2区域染色质开放程度降低;BBAi-1损害了RORγt和IL-17A表达、Th17细胞分化;BBAi-1改善EAE小鼠疾病严重程度、降低了脊髓浸润的CD4^(+)T和Th17细胞。结论BBAi-1抑制STAT3介导的增强子RORCE2染色质开放,从而降低RORγt表达和Th17分化,进而影响实验性自身免疫性脑脊髓炎(EAE)疾病进展。
The study was designed to investigate the role of BRM/BRG1 ATP inhibitor-1(BBAi-1)in STAT3-mediated RORCE2 activation.We detected the mRNA and protein levels of RORγt in splenic CD4^(+)T cells of WT and STAT3-bindingsite(BS)-/-mice.Splenic naïve CD4^(+)T cells were isolated from WT mice and cultured under Th17-polarizing condition with or without BBAi-1 for 3 days.The chromatin accessibility of RORCE2,expression of RORγt and IL-17A,and Th17 cell frequency of Th17-polarized cells were then detected.Experimental autoimmune encephalomyelitis(EAE)model was induced and divided into WT group and BBAi-1 treatment group.Clinical score of the diseased mice was recorded after immunization.On day 30 after the immunization,the mononuclear suspension and paraffin sections of spinal cord were collected and subjected to flow cytometry,HE and LFB staining,respectively.Data showed that STAT3-mediated RORCE2 activity plays a crucial role in RORγt expression and Th17 differentiation;BBAi-1 treatment resulted in decrease of RORCE2 chromatin accessibility,RORγt and IL-17A expression,Th17 cell differentiation,EAE severity,CD4^(+)T and Th17 cells infiltration in spinal cord.Taken together,BBAi-1 can inhibit STAT3-mediated RORCE2 activation,thereby reducing RORγt expression and Th17 differentiation,thus suppress the progression of EAE.
作者
王娴
董惠
周剑
杨玓
田易
韩超
吴玉章
WANG Xian;DONG Hui;ZHOU Jian;YANG Di;TIAN Yi;HAN Chao;WU Yuzhang(Department of Immunology,Medical College of Qingdao University,Qingdao 266071,China;Institute of Immunology,Third Military Medical University(Army Medical University),Chongqing 400038,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2023年第9期761-768,共8页
Immunological Journal
基金
国家自然科学基金青年项目(32100708)
国家自然科学基金面上项目(32170887)
国家自然科学基金专项(32141005)。
