α7nAChR抑制MyD88依赖性Zonulin释放改善脑梗死大鼠肠道炎症和肠屏障损伤  

α7nAChR ameliorating intestinal inflammation and intestinal barrier damage in rats with cerebral infarction by inhibiting MyD88-dependent Zonulin release

作　　者：靳子言 闫斯旸 沈紫红 陈国蕾 吴新贵[1] JIN Ziyan;YAN Siyang;SHEN Zihong;CHEN Guolei;WU Xingui(Department of Traditional Chinese Medicine,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院中医科,南宁530021

出　　处：《广西医科大学学报》2024年第2期211-217,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81360564;No.81760885);广西自然科学基金资助项目(No.2017GXNSFDA198011)。

摘　　要：目的:观察α7型烟碱乙酰胆碱受体(α7nAChR)激动剂PNU282987对脑梗死大鼠肠道炎症和肠屏障损伤的影响。方法:将36只雄性SD大鼠随机分为假手术(sham)组、中脑动脉栓塞(MCAO)组和PNU282987组。MCAO组和PNU282987组采用Longa改良线栓法制备MCAO模型,PNU282987组腹腔注射1 mg/kg PNU282987,连续7 d。苏木精—伊红(HE)染色观察肠道黏膜病理改变,酶联免疫吸附试验(ELISA)检测结肠组织肿瘤坏死因子(TNF-α)、白细胞介素(IL-6)水平及血清二胺氧化酶(DAO)、D-乳酸(DLA)、内毒素(ET)水平,免疫组化法检测结肠组织Toll样受体2(TLR2)蛋白表达,western blotting法检测结肠组织紧密连接关键蛋白咬合蛋白(Occludin)、闭合蛋白(Claudin-1)、连蛋白(Zonulin)、带状闭合蛋白(ZO-1)、TLR2、髓样分化因子88(MyD88)、核因子-κB(NF-κB)和磷酸化NF-κB(p-NF-κB)蛋白表达。结果:与sham组比较,MCAO组大鼠肠上皮脱落、肠绒毛排列紊乱及黏膜下层炎性浸润,血清DAO、DLA、ET水平及结肠组织TNF-α、IL-6含量显著增高,结肠组织Occludin、Claudin-1、ZO-1蛋白表达显著下调,Zonulin、TLR2、MyD88、p-NF-κB蛋白表达显著上调(均P<0.05)。与MCAO组比较,PNU282987组大鼠肠上皮损伤减轻,血清DAO、DLA、ET水平及结肠组织TNF-α、IL-6含量显著降低,结肠组织Occludin、Claudin-1、ZO-1蛋白表达显著上调,Zonulin、TLR2、MyD88、p-NF-κB蛋白表达显著下调(均P<0.05)。结论:PNU282987可以显著改善脑梗死大鼠肠道炎症及肠屏障损伤,其机制可能与抑制TLR2/MyD88/NF-κB信号通路有关。Objective:To observe the effect of theα7-type nicotinic acetylcholine receptor(α7nAChR)agonist PNU282987 on intestinal inflammation and intestinal barrier damage in rats with cerebral infarction.Methods:A total of 36 male SD rats were randomly divided into three groups:sham group,middle cerebral artery occlusion(MCAO)group,and PNU282987 group.MCAO group and PNU282987 group were prepared by Longa modified sature-embolus method,and PNU282987 group was intraperitoneally injected with 1 mg/kg PNU282987 for 7 consecutive days.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in intestinal mucosa.Enzyme-linked immunosorbent assay(ELISA)was performed to detect the levels of tumor necrosis factor(TNF)-αand interleukin(IL)-6 in colon tissue,and the diamine oxidase(DAO),D-lactate(DLA),endotoxin(ET)levels in serum.The expression of toll-like receptor 2(TLR2)protein in colon tissues was detected by immunohistochemistry.Western blotting was used to detect the expression of the key tight junction proteins in colon tissue,including occlusal proteins(Occludin),closed protein-1(Claudin-1),Zonulin,zona occludens(ZO-1),TLR2,myeloid differentiation factor 88(MyD88),nuclear factor-κB(NF-κB)and phosphorylated NF-κB(p-NF-κB)proteins.Results:Compared with the sham group,the MCAO group showed intestinal epithelial desquamation,disordered arrangement of intestinal villi,and inflammatory infiltration in the submucosal layer of the intestine.The levels of DAO,DLA,ET in serum,and TNF-α and IL-6 contents in colon tissue were significantly increased,while the expression of Occludin,Claudin-1,and ZO-1 proteins in colon tissue was significantly decreased,and the expression of Zonulin,TLR2,MyD88,and p-NF-κB proteins was significantly increased(all P<0.05).Compared with the MCAO group,the PNU282987 group showed reduced intestinal epithelial damage,significantly decreased levels of serum DAO,DLA,ET,and TNF-αand IL-6 contents in colon tissue,significantly increased expression of Occludin,Claudin-1,and ZO-1 proteins

关 键 词：7型烟碱乙酰胆碱受体 脑梗死 肠屏障损伤 ZONULIN 髓样分化因子88 

分 类 号：R743.3[医药卫生—神经病学与精神病学]