IL-33调控Ⅱ型固有淋巴细胞促进脓毒症腹腔巨噬细胞吞噬功能的研究  

IL-33 regulated group 2 innate lymphoid cells promoted the phagocytosis of peritoneal macrophages during sepsis

作　　者：赖登明 乐臻凯 钟晓辉 吕成杰[1] 肖轶 史波 刘智聪 章立峰[1] 刘喜旺[1] 舒强[1] Lai Dengming;Le Zhenkai;Zhong Xiaohui;Lyu Chengjie;Xiao Yi;Shi Bo;Liu Zhicong;Zhang Lifeng;Liu Xiwang;Shu Qiang(Department of Pediatric Surgery,The Children's Hospital,Zhejiang University School of Medicine,National Clinical Research Center for Child Health,Hangzhou 310052,China)

机构地区：[1]浙江大学医学院附属儿童医院儿外科,国家儿童健康与疾病临床医学研究中心,杭州310052

出　　处：《中华小儿外科杂志》2022年第7期604-609,共6页Chinese Journal of Pediatric Surgery

基　　金：国家自然科学基金(81671956, 81901989);浙江省自然科学基金项目(LY21H150005 , LY19H150005);浙江大学紫金计划资助(2022-16)。

摘　　要：目的探讨白细胞介素33 (interleukin-33, IL-33)调控腹腔Ⅱ型固有淋巴细胞(group 2 innate lymphoid cells, ILC2)募集以及ILC2对脓毒症腹腔感染时巨噬细胞的影响, 进一步了解腹腔感染的免疫调控机制。方法选择健康的6~8周雄性C57BL/6野生型小鼠40只、C57BL/6背景的Il-33-/-小鼠20只、IL-33受体ST2敲除(Il1rl1-/-)小鼠10只, 繁殖并饲养于浙江大学和美国匹兹堡大学的SPF级医学动物中心。采用盲肠结扎穿孔的方法构建脓毒症小鼠模型。接受假手术用来作为对照的小鼠除了不进行盲肠结扎穿孔操作外, 其余手术操作和脓毒症小鼠操作完全相同。将盲肠结扎穿孔模型小鼠作为脓毒症组, 将假手术操作的小鼠作为对照组。外源性腹腔注射给予IL-33蛋白, 流式细胞术检测小鼠腹腔灌洗液中ILC2的变化及其胞内细胞因子水平。运用体外实验证实ILC2分泌的IL-9和IL-13对巨噬细胞吞噬能力的影响。采用t检验或One-Way ANOVA检验统计分析。结果①相较于ILC2在IL-33刺激之前和之后, ILC2在腹腔Lin-CD45+细胞中的比例分别为9.90±0.80比18.43±0.46, ILC2受刺激后在腹腔中的比例增加, 差异具有统计学意义(P<0.001), ILC2在腹腔中的数量分别为(5 786.00±289.50)个比(17 820.00±324.30)个, ILC2受刺激后在腹腔中的数量增加, 差异具有统计学意义(P<0.001)。②C57BL/6野生型小鼠在接受构建脓毒症小鼠模型操作24 h后腹腔ILC2募集较对照组显著增多, 20.50±0.59比10.83±1.01, 差异具有统计学意义(P=0.001);Il-33-/-、Il1rl1-/-小鼠在接受构建脓毒症小鼠模型操作24 h后腹腔中的ILC2并未增加, 而给对照组小鼠腹腔注射重组IL-33蛋白后ILC2的比例比未注射的对照组Il-33-/-小鼠的ILC2比例显著增多, 10.53±0.47比19.43±0.87, 差异具有统计学意义(P<0.001);给脓毒症组小鼠腹腔注射重组IL-33蛋白后ILC2的比例比未注射的脓毒症Il-33-/-小鼠的ILC2比例显著增多, 10.13±0.57比2Objective To explore the regulation of group 2 innate lymphoid cells(ILC2)recruitment by interleukin-33(IL-33),to examine the effect of ILC2 on macrophages during peritoneal infection in sepsis and to further elucidate the immune regulatory mechanism of peritoneal infection.Methods Forty healthy 6 to 8 week-old male C57BL/6 wild-type mice,twenty Il-33-/-mice on a C57BL/6 background,and ten IL-33 receptor ST2 knockout(Il1rl1-/-)mice were bred and housed in the SPF Medical Animal Center of Zhejiang University and the University of Pittsburgh,USA.Receipts of sham surgery were designated as controls and underwent identical surgical procedures were septic mice.IL-33 protein was administered to rmIL-33 group via an exogenous intraperitoneal injection and the changes of ILC2 and its intracellular cytokine levels in peritoneal lavage fluid were detected by flow cytometry.The effects of IL-9 and IL-13 secreted by ILC2 on phagocytic ability of macrophages were confirmed by in vitro experiments.Statistical analysis was performed with t-test or one-way ANOVA test.Results Compared with ILC2 before/after IL-33 stimulation,the proportion of ILC2 in Lin-CD45+cells of abdominal cavity increased[(9.90±0.80)vs.(18.43±0.46)]and the proportion of ILC2 in abdominal cavity jumped after IL-33 stimulation and the difference was statistically significant(P<0.001).The number of ILC2 in abdominal cavity was[(5786.00±289.50)vs.(17820.00±324.30)]and the number of ILC2 spiked in abdominal cavity after IL-33 stimulation and the difference was statistically significant(P<0.001).C57BL/6 wild-type mice had a marked increase of peritoneal ILC2 recruitment as compared with control group at 24 h postoperatively[(20.50±0.59)vs.(10.83±1.01)]and the difference was statistically significant(P=0.001).However,ILC2 in peritoneal cavity of Il-33-/-and Il1rl1-/-mice showed no change at 24 h postoperatively.Moreover,the proportion of ILC2 in Il-33-/-mice were significantly increased after IL-33 stimulation compared to non-stimulation group(10.53±0.47 vs

关 键 词：脓毒症 腹腔 Ⅱ型固有淋巴细胞 白介素33 巨噬细胞 

分 类 号：R720.597[医药卫生—急诊医学]