高脂饲喂小鼠结肠ILC3s变化对肠屏障应激能力的影响  

Effects of colonic ILC3s alterations on intestinal barrier stress capability in mice fed with high-fat diet

作　　者：徐凤华 喻丽玲 路晓光[3] 王迎莉[4] 宋轶[3] Xu Fenghua;Yu Liling;Lu Xiaoguang;Wang Yingli;Song Yi(Graduate School of Dalian University,Dalian 116622,China;Qionghai People’s Hospital of Hainan Province,Qionghai 571499,China;Department of Emergency Medicine,the Affiliated Zhongshan Hospital of Dalian University,Dalian 116001,China;Emergency ICU,the Affiliated Zhongshan Hospital of Dalian University,Dalian 116001,China)

机构地区：[1]大连大学研究生学院,辽宁大连116622 [2]海南省琼海市人民医院,海南琼海571499 [3]大连大学附属中山医院急诊科,辽宁大连116001 [4]大连大学附属中山医院急诊ICU,辽宁大连116001

出　　处：《中华卫生应急电子杂志》2024年第2期107-114,共8页Chinese Journal of Hygiene Rescue(Electronic Edition)

基　　金：国家自然科学基金资助项目(81673801、81473512)。

摘　　要：目的探讨高脂饲喂小鼠结肠三型固有淋巴细胞(ILC3s)的变化对肠黏膜屏障应激能力影响的潜在机制。方法将24只SPF级雄性C57BL/6J小鼠适应性饲养1周后,随机分为空白对照组(N-CON组)、重症急性胰腺炎(SAP)对照组(N-SAP组)、空白高脂组(H-CON组)、SAP高脂组(H-SAP组),每组6只。N-CON、N-SAP给予普通饮食,H-CON、H-SAP给予高脂饮食,持续21周;N-SAP组、H-SAP组采用腹腔注射20%左旋精氨酸(L-Arg)作为应激刺激制备小鼠SAP模型,N-CON组、H-CON组予以腹腔注射等体积等渗盐水。造模48 h后,酶联免疫吸附试验(ELISA)检测甘油三酯(TG)、总胆固醇(TC)、血清淀粉酶(AMS)、脂多糖(LPS)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平;HE染色观察胰腺、肺、结肠组织病理学形态;流式细胞术分析结肠ILC3s占淋巴细胞比例和IL-22+ILC3s占结肠ILC3s比例;免疫荧光法检测结肠荆豆凝集素(UEA-1)、白细胞介素-22受体A1(IL-22RA1)表达情况;Western blot检测岩藻糖基转移酶2(Fut2)、IL-22RA1蛋白的表达水平。结果与N-CON组相比,H-CON组小鼠体重、TG、TC以及血清AMS、LPS、IL-6、TNF-α水平显著升高(P<0.05),结肠ILC3s占淋巴细胞比例显著下降(P<0.05),IL-22+ILC3s占结肠ILC3s比例升高(P>0.05),Fut2、IL-22RA1蛋白表达下降(P>0.05),UEA-1、IL-22RA1平均荧光强度略有下降(P>0.05);给予20%Arg应激刺激后,与N-CON组相比,N-SAP组血清AMS、LPS、IL-6、TNF-α水平显著升高(P<0.05),结肠ILC3s占淋巴细胞比例显著下降(P<0.05),IL-22+ILC3s占结肠ILC3s比例变化显著升高(P<0.05),Fut2、IL-22RA1蛋白表达升高(P>0.05),UEA-1平均荧光强度显著下降(P<0.05),IL-22RA1平均荧光强度升高(P>0.05);与N-SAP组相比,H-SAP组结肠ILC3s占淋巴细胞比例降低(P>0.05),IL-22+ILC3s占结肠ILC3s比例升高(P>0.05),UEA-1平均荧光强度下降(P>0.05),IL-22RA1平均荧光强度显著升高(P<0.05),Fut2、IL-22RA1蛋白表达增加(P>0.05)。结论高脂饲喂小鼠的肠黏�Objective To investigate the impact of colonic group 3 innate lymphoid cells(ILC3s)on intestinal barrier stress capability in mice fed with high-fat diet.Methods Twenty-four SPF male C57BL/6J mice were adaptively fed for 1 week and then randomly divided into 4 groups(n=6):blank control group(N-CON),SAP control group(N-SAP),high-fat diet control group(H-CON),and SAP high-fat diet group(H-SAP).N-CON and N-SAP received a normal diet,while H-CON and H-SAP were fed a high-fat diet for 21 weeks.In the SAP group,20%Arg was injected intraperitoneally as stress stimulation to prepare the mouse SAP model,while an equal volume of saline was injected into the CON groups.After 48 hours of modeling,serum levels of TC,TG,AMY,LPS,TNF-α,IL-6 were measured by ELISA;pancreatic histopathology as well as lung and colon tissue morphology were observed using HE staining;flow cytometry was employed to analyze changes in the proportion of colonic ILC3s among lymphocytes and changes in the proportion of IL-22+ILC3s among colonic ILC3s;immunofluorescence was conducted to detect UEA-Ⅰand IL-22RA1 expression;Western blot was performed to measure Fut2 and IL-22RA1 protein expression levels.Results Compared with the N-CON group,the H-CON group exhibited significantly increased levels of body weight,TG,TC,serum AMS,LPS,IL-6 and TNF-α(P<0.05),along with a significant decrease in the proportion of colon ILC3s in lymphocytes(P<0.05).The proportion of IL-22+ILC3s in the colon showed an increase(P>0.05),while the expressions of Fut2 and IL-22RA1 were decreased(P>0.05).The average fluorescence intensity of UEA-1 and IL-22RA1 slightly decreased(P>0.05).After stimulation with a 20%Arg stress in the N-SAP group compared to the N-CON group,there was a significant increase in serum AMS,LPS,IL-6 and TNF-a levels(P<0.05)and a significant decrease in the proportion of colon ILC3s in lymphocytes(P<0.05).The proportion of IL-22+ILC3s in the colon significantly increased(P<0.05),while protein expression of Fut2 and IL-22RA1 was increased as well(P>0.05).The

关 键 词：ILC3s 高脂饮食 Fut2 肠黏膜屏障 应激能力 

分 类 号：R574[医药卫生—消化系统]