右美托咪定对自发性结肠炎小鼠肠黏膜上皮屏障功能的影响  

Influence of dexmedetomidine on intestinal mucosal epithelial barrier function in spontaneous colitis mice

作　　者：邢海林 胡浩然 夏勇[3] 王宏刚[3] Xing Hailin;Hu Haoran;Xia Yong;Wang Honggang(Department of Anesthesiology,Taizhou People's Hospital,Nanjing Medical University,Taizhou 225300,China;Postgraduate Training Base of Dalian Medical University,Taizhou People's Hospital,Taizhou 225300,China;Department of General Surgery,Taizhou People's Hospital,Nanjing Medical University,Taizhou 225300,China)

机构地区：[1]南京医科大学附属泰州人民医院(泰州市人民医院)麻醉科,泰州225300 [2]大连医科大学研究生培养基地(泰州市人民医院),泰州225300 [3]南京医科大学附属泰州人民医院(泰州市人民医院)普通外科,泰州225300

出　　处：《中华炎性肠病杂志（中英文）》2022年第3期240-247,共8页Chinese Journal of Inflammatory Bowel Diseases

基　　金：国家自然科学基金(81600434);江苏省卫生健康委员会面上项目(M2020084)。

摘　　要：目的探讨右美托咪定(DEX)对自发性结肠炎小鼠肠黏膜上皮屏障功能的影响及相关机制。方法采用白细胞介素-10基因敲除(IL-10^(-/-))小鼠作为自发性结肠炎模型。将IL-10^(-/-)小鼠随机分为模型组(IL-10^(-/-)组)和治疗组(DEX组),取野生型小鼠作为对照,设为WT组。DEX组小鼠每天1次给予20μg/kg的DEX腹腔注射,IL-10^(-/-)组和WT组小鼠给予等体积的PBS腹腔注射,干预14 d。计算3组小鼠疾病活动度(DAI)、体质量变化和结肠病理组织学评分,采用流式细胞术检测黏膜固有层CD4^(+)CD45^(+)T细胞的比例,酶联免疫吸附试验检测小鼠结肠黏膜炎症因子(IFN-γ和IL-17)的表达,Ussing Chamber检测结肠黏膜甘露醇通透性和肠上皮细胞跨膜电阻(TER),共聚焦显微镜检测肠上皮细胞紧密连接蛋白(Occludin和ZO-1)的表达,Western blot检测Occludin、ZO-1和凋亡通路蛋白(TNF-α和TNFR2)的表达,Tunel染色检测结肠细胞凋亡,荧光定量PCR检测TNF-α和TNFR2 mRNA的表达。统计分析3组间上述指标的差异。结果(1)炎症程度:与WT组比较,IL-10^(-/-)组小鼠DAI评分明显更高,病理组织学评分更高,CD4^(+)CD45^(+)T细胞比例更高,IFN-γ和IL-17的表达更高,差异均有统计学意义(均P<0.05)。与IL-10^(-/-)组比较,DEX组小鼠DAI评分明显更低,病理组织学评分更低,CD4^(+)CD45^(+)T细胞比例更低,IFN-γ和IL-17的表达更低,差异均有统计学意义(均P<0.05)。(2)通透性:与WT组比较,IL-10^(-/-)组小鼠结肠甘露醇通透性更高,TER更低,Occludin和ZO-1蛋白的表达量更低,差异均有统计学意义(均P<0.05),并且蛋白表达的位置发生迁移。与IL-10^(-/-)组比较,DEX组甘露醇通透性明显更低,TER更高,Occludin和ZO-1的表达量更高,差异均有统计学意义(均P<0.05),蛋白表达位置也发生明显的纠正。(3)凋亡:与WT组比较,IL-10^(-/-)组小鼠上皮细胞凋亡更多,结肠黏膜TNF-α和TNFR2蛋白表达更高,TNF-α和TNFR2的mRNA表达更高,差异均Objective To investigate the influence of dexmedetomidine(DEX)on intestinal mucosal epithelial barrier function in spontaneous colitis mice.Methods IL-10 gene knockout(IL-10^(-/-))mouse was set as the model of spontaneous colitis.IL-10^(-/-)mice were randomly divided into model group(IL-10^(-/-)group)and treatment group(DEX group).The wild type mice(WT group)were set as control.The mice in DEX group was given intraperitoneal injection of 20μg/kg DEX once a day for 14 days.The mice in IL-10^(-/-)group and WT group were given the same volume of PBS intraperitoneal injection for 14 days.The disease activity index(DAI),the weight change and colon histopathological score were caculated.The ratio of CD4^(+)CD45^(+)T cells in lamina propria of mucosa was detected by flow cytometry.The expression of inflammatory factors including interferon-γ(IFN-γ)and IL-17 was detected by enzyme linked immunosorbent assay(ELISA).The mannitol permeability and transepithelial electrical resistance(TER)of colonic mucosa were detected by Ussing Chamber.The expression of tight junction proteins(Occludin and ZO-1)in intestinal epithelial cells was detected by confocal microscopy.The protein expression of Occludin,ZO-1,tumor necrosis factor-α(TNF-α)and tumor necrosis factor receptor 2(TNFR2)was detected by Western blot.The colon cell apoptosis was detected by Tunel staining.The mRNA expression of TNF-α and TNFR2 was detected by fluorescence quantitative PCR.The differences of above indexes between the three groups were analyzed statistically.Results(1)Inflammation:Compared with WT group,the DAI,histopathological score,ratio of CD4^(+)CD45^(+)T cells,expression of IFN-γ and IL-17 were higher in IL-10^(-/-)group(all P<0.05).Compared with IL-10^(-/-)group,the DAI,histopathological score,ratio of CD4^(+)CD45^(+)T cells,expression of IFN-γ and IL-17 were lower in DEX group(all P<0.05).(2)Permeability:Compared with WT group,the mannitol permeability was higher,TER was lower,the protein expression of Occludin and ZO-1 was lower in IL-10^(-/-

关 键 词：结肠炎 自发性 右美托咪定 基因 白细胞介素10 基因敲除 通透性 凋亡 小鼠 

分 类 号：R614[医药卫生—麻醉学]