摘要
目的观察肝硬化合并自发性细菌性腹膜炎(SBP)患者外周血和腹水中固有淋巴样细胞(ILC)亚群的变化。方法收集2019年11月至2020年11月在驻马店市中心医院住院的62例肝硬化患者的病例资料,其中肝硬化合并单纯腹水41例(单纯腹水组),肝硬化合并SBP 21例(SBP组),同时收集同期进行健康查体的对照者20名(健康对照组)。所有患者和对照者采集外周血,分离外周血单个核细胞(PBMC),肝硬化患者常规穿刺获取腹水,分离腹水单个核细胞,流式细胞术检测PBMC和腹水单个核细胞中ILC1、ILC2、ILC3亚群的比例变化。分选腹水中CD3^(-)CD19^(-)CD20^(-)CD14^(-)细胞(即lin^(-)细胞),使用脂多糖(LPS)刺激培养24 h,实时定量PCR法检测lin^(-)细胞中转录因子T-bet、GATA3和RORγt mRNA相对表达量,酶联免疫吸附试验检测培养上清中细胞因子水平。比较各组间外周血和腹水中ILC亚群的区别。结果单纯腹水组男29例、女12例,年龄M(Q_(1),Q_(3))为49(33,78)岁,SBP组男12例、女9例,年龄50(37,76)岁,健康对照组男11名、女9名,年龄48(32,69)岁。三组外周血和腹水中均可检测到lin^(-)CD45^(+)CD161^(+)CD127^(+)ILC细胞,总ILC细胞占PBMC的比例在单纯腹水组、SBP组及健康对照组之间的差异无统计学意义(P=0.235)。总ILC占腹水单个核细胞的比例在单纯腹水组及SBP组之间的差异亦无统计学意义(P=0.232)。CD117^(-)CRTh2^(-)ILC1、CRTh2^(+)ILC2、CD117^(+)CRTh2^(-)ILC3占外周血ILC的比例在单纯腹水组、SBP组及健康对照组之间的差异均无统计学意义(均P>0.05)。SBP组ILC1占腹水ILC的比例高于单纯腹水组(35.69%±3.39%比26.40%±3.85%,P<0.001),而ILC2占腹水ILC的比例低于单纯腹水组(36.83%±7.70%比48.35%±9.45%,P<0.001),ILC3占腹水ILC的比例在两组间的差异无统计学意义(P=0.230)。SBP组腹水lin-细胞经LPS刺激后T-bet mRNA相对表达量和干扰素(IFN)-γ分泌水平均高于单纯腹水组(均P<0.001),GATA3 mRNA相对�
Objective To investigate the change of innate lymphoid cells(ILC)subsets in peripheral blood and ascites in liver cirrhotic patients complicated with spontaneous bacterial peritonitis(SBP).Methods The data of 62 patients with liver cirrhosis admited to the Zhumadian Central Hospital from November 2019 to November 2020 were analyzed.Among them,41 cases were complicated with untainted ascites(untainted ascites group),while the other 21 cases were complicated with SBP(SBP group).Meanwhile,20 cases of controls who received healthy examination in the same period were also enrolled(control group).Peripheral blood mononuclear cell(PBMC)was isolated from peripheral blood of all patients and controls.Mononuclear cell in ascites was isolated from patients with liver cirrhosis.The percentage of ILC1,ILC2,and ILC3 subsets in PBMC and mononuclear cell in ascites were measured by flow cytometry.CD3^(-)CD19^(-)CD20^(-)CD14^(-)cells(lin^(-)cells)were purified from ascites and were stimulated with lipopolysaccharide(LPS)for 24 h.The transcription factor T-bet,GATA3,and RORγt mRNA relative level in lin-cells was semi-quantified by real-time PCR.Cytokine level in the supernatants was measured by enzyme linked immunosorbent assay.Differences of ILC subsets in peripheral blood and ascites were compared among groups.Results There were twenty-nine males and twelve females in untainted ascites group,aged M(Q_(1),Q_(3))49(33,78)years.There were twelve males and nine females in SBP group,aged 50(37,76)years.There were eleven males and nine females in control group,aged 48(32,69)years.lin-CD45^(+)CD161^(+)CD127^(+)ILC cells could be detected in both peripheral blood and ascites.There was no significant difference in total ILC percentage within PBMC among untainted ascites group,SBP group,and control group(P=0.235).There was also no significant difference of total ILC percentage within mononuclear cells in ascites between untainted ascites group and SBP group(P=0.232).The differences were not statistically significant of peripheral CD117^(
作者
陈攀丽
张炳勇
付雪琴
张逸强
谢华平
Chen Panli;Zhang Bingyong;Fu Xueqin;Zhang Yiqiang;Xie Huaping(Department of Gastroenterology,Zhumadian Central Hospital,Zhumadian 463000,China;Department of Gastroenterology,Henan Provincial People′s Hospital,Zhengzhou 450003,China;Department of Gastroenterology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)
出处
《中华医学杂志》
CAS
CSCD
北大核心
2022年第2期141-146,共6页
National Medical Journal of China
关键词
肝硬化
腹水
腹膜炎
固有淋巴样细胞
Liver cirrhosis
Peritonitis
Ascites
Innate lymphoid cells
