3种经典急性胰腺炎模型中回肠和胰腺抗菌肽的动态变化  

Dynamic Changes of Antimicrobial Peptides in Ileum and Pancreas of Three Classical Acute Pancreatitis Models

作　　者：黄慧珍 高玮[2] 殷诺铭 黄春兰[2,3] 梅启享 曾悦 HUANG Huizhen;GAO Wei;YIN Nuoming;HUANG Chunlan;MEI Qixiang;ZENG Yue(Shanghai General Hospital of Nanjing Medical University,Shanghai,200080;Department of Gastroenterology;Shanghai Key Laboratory of Pancreatic Disease,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai)

机构地区：[1]南京医科大学附属上海一院临床医学院,200080 [2]上海交通大学医学院附属第一人民医院消化内科 [3]上海市胰腺疾病重点实验室

出　　处：《胃肠病学》2024年第2期75-85,共11页Chinese Journal of Gastroenterology

基　　金：国家自然科学基金面上项目(82270671);国家自然科学基金青年科学基金项目(82200714,82300731);上海交通大学医学院附属第一人民医院特色研究项目(CCTR⁃2022B02)。

摘　　要：背景:肠道菌群紊乱以及肠道屏障受损导致的细菌移位参与了急性胰腺炎(AP)的重症化进程。抗菌肽参与调节肠道菌群,但其在AP病程中的变化和作用尚不明确。目的:探究3种经典AP模型中回肠和胰腺抗菌肽的动态变化和意义。方法:构建雨蛙肽+脂多糖(CAE+LPS)、牛磺胆酸钠(N⁃Tau)和L⁃精氨酸(L⁃Arg)AP小鼠模型,评估胰腺和回肠组织病理学变化,以real⁃time PCR检测促炎细胞因子和抗菌肽,包括溶菌酶(LZM)、分泌型磷脂酶A2(sPLA2)、血管生成素4(Ang4)、再生胰岛衍生蛋白3(REG3)家族、β防御素家族、cathelicidin相关抗菌肽(CRAMP)、糖蛋白2(GP2)在回肠和(或)胰腺组织中的表达,分析抗菌肽表达与胰腺、回肠病理损伤的相关性。结果:3组AP模型组胰腺和回肠组织在各时间点均可观察到不同程度的病理损伤;在72 h内,CAE+LPS和N⁃Tau模型组病理损伤达峰值后开始减轻,L⁃Arg模型组病理损伤则逐渐加重。与相应对照组相比,3组AP模型组回肠组织LZM、sPLA2、Ang4和胰腺组织CRAMP、GP2、β防御素mRNA表达整体上显著下调(P均<0.05),CAE+LPS和N⁃Tau模型组呈先下降后回升趋势,L⁃Arg模型组则持续下降;回肠组织REG3β、REG3γmRNA表达在48 h或24 h显著上调达峰值后于72 h显著下调(P均<0.05),胰腺组织两者表达整体上显著上调(P均<0.05),但CAE+LPS和N⁃Tau模型组72 h时已回落;回肠组织β防御素mRNA表达在病程早期(12 h)显著上调(P均<0.05)后逐渐下降。Spearman相关系数分析表明,3种造模方式下,回肠LZM、sPLA2、Ang4和胰腺CRAMP、GP2、β防御素表达与胰腺、回肠病理损伤呈显著负相关(P均<0.05),胰腺REG3β表达则与胰腺、回肠病理损伤呈显著正相关(P均<0.05)。结论:在3种经典AP模型中,回肠LZM、sPLA2、Ang4和胰腺CRAMP、GP2、β防御素家族、REG3β表达均随AP病情严重程度呈动态变化,回肠和胰腺抗菌肽可能通过调节肠道微生态影响AP时的胰�Background:Intestinal microbiota dysbiosis and impaired intestinal barrier,leading to bacterial translocation,are involved in acute pancreatitis(AP)exacerbation.Antimicrobial peptides participate in the regulation of intestinal microbiota,yet their changes and roles in the course of AP are unclear.Aims:To investigate the dynamic changes and significance of antimicrobial peptides in the ileum and pancreas among three classical AP models.Methods:Three AP mouse models were established by using cathelicidin plus lipopolysaccharide(CAE+LPS),sodium taurocholate(N⁃Tau)and L⁃arginine(L⁃Arg),respectively.The pathological changes of pancreatic and ileal tissues were observed and scored.Real⁃time PCR was applied to detect the expression levels of the proinflammatory cytokines,and antimicrobial peptides including lysozyme(LZM),secretory phospholipase A2(sPLA2),angiogenin 4(Ang4),regenerating islet⁃derived protein 3(REG3)family,β⁃defensin family,cathelicidin⁃related antimicrobial peptide(CRAMP),and glycoprotein 2(GP2)in ileum and/or pancreas.The association between expressions of antimicrobial peptides and the injuries of pancreas and ileum was analyzed.Results:Pancreatic and ileal injuries could be observed in all three AP models in different time points with various degrees.The pathological scores of the CAE+LPS and N⁃Tau models reached the highest level and then declined from 0⁃72 h,while those of L⁃Arg model progressively increased within 72 hours.Compared with the corresponding controls,the mRNA levels of LZM,sPLA2,and Ang4 in ileal tissue,and the mRNA levels of CRAMP,GP2,andβ⁃defensins in pancreatic tissue,were generally downregulated in all three AP models(all P<0.05).CAE+LPS and N⁃Tau models showed a trend of initial decrease followed by partial recovery,while L⁃Arg model exhibited a gradual downregulation trend.The mRNA levels of REG3βand REG3γin ileum upregulated and reached the peak at 48 h or 24 h and downregulated significantly at 72 h in all three AP models(all P<0.05);while in panc

关 键 词：急性胰腺炎 肠道微生态 抗菌肽 疾病模型 动物 

分 类 号：R576[医药卫生—消化系统]