摘要
目的探讨降钙素基因相关肽(CGRP)对脓毒症大鼠肠黏膜屏障的保护作用及机制。方法30只SD雄性大鼠随机分为正常组、假手术组、脓毒症组(CLP组)、CGRP治疗组(CGRP组)和抗CGRP组,每组6只。采用盲肠结扎穿孔制备脓毒症大鼠模型,CGRP组大鼠造模前经尾静脉注射0.5 ml CGRP(30μg/kg),抗CGRP组大鼠造模前经尾静脉依次注射0.5 ml CGRP(30μg/kg)和0.5 ml CGRP8-37(30μg/kg)。检测各组大鼠术前0.5 h及术后48 h的血清二胺氧化酶(DAO)、D-乳酸水平;HE染色法观察小肠组织病理学变化;TUNEL法检测小肠组织凋亡情况;免疫组化法检测小肠组织中磷酸化Bcl-xl/Bcl-2相关死亡启动子(p-BAD)、磷酸化细胞外调节蛋白激酶(p-ERK)和caspase-3的表达。结果与正常组比较,CLP组大鼠小肠组织病理损害明显,凋亡增加,p-ERK、p-BAD表达减少,caspase-3表达增加,血清DAO、D-乳酸水平明显升高(P<0.05);与CLP组比较,CGRP组大鼠小肠组织病理损害明显减轻,凋亡减少,p-ERK、p-BAD表达增加,caspase-3表达减少,血清DAO、D-乳酸水平明显降低(P<0.05);与CGRP组比较,抗CGRP组大鼠小肠组织病理损害明显加重,凋亡增加,p-ERK、p-BAD表达减少,caspase-3表达增加,血清DAO、D-乳酸水平明显升高(P<0.05)。结论脓毒症大鼠小肠黏膜通透性及组织凋亡增加,上调的p-ERK和p-BAD证实CGRP可减少小肠组织凋亡,从而发挥保护肠黏膜屏障的作用。
Objective To investigate the mechanism of calcitonin gene-related peptide(CGRP)in protecting intestinal barrier integrity by reducing intestinal apoptosis in sepsis rats.Methods Thirty SD male rats were randomized into 5 groups(n=6):Normal group,sham group,sepsis group(CLP group),CGRP treatment group(CGRP group)and anti-CGRP group.Rats were injected with 0.5 ml CGRP(30μg/kg)in the CGRP group and injected with 0.5 ml CGRP(30μg/kg)and 0.5 ml CGRP8-37(30μg/kg)in the anti-CGRP group.Serum diamine oxidase(DAO)and D-lactic acid levels were measured 30 minutes before the operation and 48 hours after the modeling.Histopathological changes of the small intestine were observed by HE staining;apoptosis of small intestine was detected by TUNEL assay;the expression of phosphorylated Bcl-xl/Bcl-2 related death promoter(p-BAD),phosphorylated extracellular signalregulated kinase(p-ERK),caspase-3 were observed by immunohistochemistry.Results Compared with the normal group,the small intestine tissues were significantly damaged in the CLP group,with increased apoptosis,decreased p-ERK and p-BAD expressions,increased caspase-3 expressions(P<0.05).Meanwhile,serum DAO and D-lactic acid levels significantly increased(P<0.05).The damage of the small intestine in the CGRP group was significantly reduced compared with the CLP group,with decreased apoptosis and caspase-3 expression,increased p-ERK and p-BAD expression and significantly decreased serum DAO and D-lactic acid levels(P<0.05).Compared with the CGRP group,the small intestine damage was significantly aggravated in the anti-CGRP group,with increased apoptosis,decreased p-ERK and p-BAD expressions,increased caspase-3 expressions,and significantly increased serum DAO and D-lactic acid levels(P<0.05).Conclusion Intestinal mucosal permeability and apoptosis increase in sepsis rats.CGRP protects the integrity of the intestinal mucosal barrier through reduced intestinal tissue apoptosis with increased p-ERK and p-BAD.
作者
郭久冰
王隽笙
林锦洲
孙家伟
张道建
徐文军
陈亚阳
徐志鹏
丰威
苏磊
刘慧恒
GUO Jiu-bing;WANG Jun-sheng;LIN Jin-zhou;SUN Jia-wei;ZHANG Dao-jian;XU Wen-jun;CHEN Ya-yang;XU Zhi-peng;FENG Wei;SU Lei;LIU Hui-heng(Second Department of General Surgery,Xiamen Haicang Hospital,Xiamen,Fujian 361000,China;Department of Emergency,Zhongshan Hospital Affiliated to Xiamen University,Xiamen,Fujian 361000,China;Department of Respiratory Medicine,Shishi Municipal Hospital,Shishi,Fujian 362700,China;Department of Intensive Care Unit,key Laboratory of Hot Zone Trauma Care and Tissue Repair of Chinese PLA,General Hospital of Southern Theatre Command,Guangzhou 510030,China)
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2019年第12期1024-1029,共6页
Medical Journal of Chinese People's Liberation Army
基金
福建省医学创新课题(2015-CXB-50)~~
关键词
脓毒症
肠黏膜屏障
降钙素基因相关肽
sepsis
intestinal mucosal barrier
calcitonin gene-related peptide
