摘要
目的探讨组蛋白甲基转移酶果蝇zeste基因增强子同源物2(EZH2)调控脓毒症T淋巴细胞功能失调的作用及机制。方法按随机数字表法将24只雄性C57BL/6小鼠分为假手术组、脓毒症模型组〔盲肠结扎穿孔术(CLP)+二甲基亚砜(DMSO)组〕及EZH2选择性抑制剂干预组(CLP+GSK126组),每组8只。采用CLP法制备脓毒症小鼠模型;假手术组小鼠不结扎穿刺盲肠,其余操作同CLP+DMSO组。CLP+DMSO组及CLP+GSK126组小鼠术后立即分别腹腔注射DMSO或GSK126(10 mg/kg)。术后24 h处死小鼠取肠系膜淋巴结,采用流式细胞仪检测T淋巴细胞EZH2表达、T淋巴细胞凋亡率、细胞增殖标志物ki-67抗原阳性T淋巴细胞(ki-67^(+))比例、γ-干扰素阳性T淋巴细胞(IFN-γ^(+))比例、程序性死亡受体-1阳性T淋巴细胞(PD-1+)比例及程序性死亡配体-1阳性T淋巴细胞(PD-L1^(+))比例。结果与假手术组相比,CLP+DMSO组小鼠肠系膜淋巴结T淋巴细胞EZH2表达升高。与CLP+DMSO组相比,CLP+GSK126组小鼠肠系膜淋巴结CD3+比例升高(0.70±0.02比0.50±0.07,P<0.01),表明抑制EZH2可增加脓毒症小鼠淋巴结T淋巴细胞数量;同时,CLP+GSK126组淋巴结CD4^(+)及CD8^(+)细胞中ki-67^(+)比例显著上升(CD4^(+):0.74±0.05比0.63±0.04,CD8^(+):0.82±0.06比0.70±0.04,均P<0.05),表明抑制EZH2可增加脓毒症小鼠淋巴结增殖活性较高的T淋巴细胞比例;另外,CLP+GSK126组小鼠肠系膜淋巴结T淋巴细胞凋亡率与CLP+DMSO组比较差异无统计学意义〔CD4^(+):(21.53±2.87)%比(20.48±3.21)%,CD8^(+):(8.34±1.02)%比(7.71±1.38)%,均P>0.05〕,表明抑制EZH2对脓毒症小鼠淋巴结T淋巴细胞凋亡无明显影响。与CLP+DMSO组相比,CLP+GSK126组小鼠肠系膜淋巴结IFN-γ^(+)CD4^(+)、IFN-γ^(+)CD8^(+)比例也显著增加(IFN-γ^(+)CD4^(+):0.31±0.11比0.14±0.06,IFN-γ^(+)CD8^(+):0.30±0.10比0.13±0.06,均P<0.05),表明抑制EZH2可增强脓毒症小鼠淋巴结T淋巴细胞IFN-γ分泌能力;同时,CLP+GSK126组小鼠肠系膜�
Objective To investigate the effect and mechanism of histone methyltransferase enhancer of zeste homolog 2(EZH2)on sepsis-induced T cell dysfunction.Methods Twenty-four male C57BL/6 mice were divided into three groups randomly:sham operated group,sepsis model group[cecum ligation and puncture(CLP)+dimethyl sulfoxide(DMSO)group]and EZH2 selective inhibitor treated group(CLP+GSK126 group),with 8 mice in each group.Sepsis murine model was reproduced by CLP.CLP+DMSO group and CLP+GSK126 group were treated with DMSO or GSK126(10 mg/kg)respectively right after surgery through intraperitoneal injection.The mice were sacrificed 24 hours after operation,and the mesenteric lymph nodes were collected.The expression of EZH2,apoptosis rates,cell proliferation marker ki-67 antigen positive T lymphocytes(ki-67^(+)cell),interferon-γpositive T lymphocytes(IFN-γ^(+)cell),programmed death receptor-1 positive T lymphocytes(PD-1+cell)and programmed death-ligand 1 positive T lymphocytes(PD-L1^(+)cell)were determined by flow cytometry.Results Compared with sham operated group,the expression of EZH2 in T lymphocytes was up-regulated on mesenteric lymph nodes of CLP+DMSO group.Compared with CLP+DMSO group,the ratio of CD3+T lymphocytes in CLP+GSK126 group was up-regulated(0.70±0.02 vs.0.50±0.07,P<0.01),indicating that the EZH2 inhibitor could increase the number of T lymphocytes in lymph nodes of septic mice;the ratio of ki-67^(+)cells in CD4^(+)and CD8^(+)T lymphocytes in CLP+GSK126 group was increased(CD4^(+):0.74±0.05 vs.0.63±0.04,CD8^(+):0.82±0.06 vs.0.70±0.04,both P<0.05),indicating that the EZH2 inhibitor could increase the ratio of T lymphocytes with high proliferative activity in lymph nodes of septic mice.However,no significant difference was found on both CD4^(+)and CD8^(+)T lymphocytes apoptosis rates in the mesenteric lymph nodes of mice between CLP+GSK126 group and CLP+DMSO group[CD4^(+):(21.53±2.87)%vs.(20.48±3.21)%,CD8^(+):(8.34±1.02)%vs.(7.71±1.38)%,both P>0.05],indicating that no extra T lymphocytes apoptosis
作者
李哲
赵冬旸
周晓慧
唐伦先
Li Zhe;Zhao Dongyang;Zhou Xiaohui;Tang Lunxian(Department of Emergency Medicine and Critical Care,East Hospital,Tongji University,Shanghai 200120,China;Tongji University School of Medicine,Shanghai 200120,China;Translational Medicine Research Center,East Hospital,Tongji University,Shanghai 200120,China)
出处
《中华危重病急救医学》
CAS
CSCD
北大核心
2022年第4期357-361,共5页
Chinese Critical Care Medicine
基金
国家自然科学基金(81970072)
上海市科技计划项目(20Y11901200)
上海市浦东新区医药卫生领先人才培养计划项目(PWRI2019-05)。
