5-羟色胺4受体激动剂对糖尿病小鼠结肠黏膜巨噬细胞极化的影响  

Effects of 5-HT_(4)R agonist on macrophage polarization in colonic mucosa of diabetic mice

作　　者：寇玥婷 成颖莹 王娟 戎伟芳 张国花 KOU Yue-ting;CHENG Ying-ying;WANG Juan;RONG Wei-fang;ZHANG Guo-hua(Dept.of Anatomy and Physiology,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China)

机构地区：[1]上海交通大学医学院解剖学与生理学系,上海200025

出　　处：《同济大学学报（医学版）》2021年第4期453-458,466,共7页Journal of Tongji University(Medical Science)

基　　金：国家自然基金面上项目(81770533)。

摘　　要：目的研究5-羟色胺4受体(5-hydroxytryptamine 4 receptor,5-HT_(4)R)激动剂对糖尿病结肠黏膜巨噬细胞极化的影响。方法使用链脲佐菌素(streptozotocin,STZ)建立小鼠1型糖尿病模型,使用5-HT_(4)R激动剂RS67333进行治疗干预,实验动物随机分为3组,即非糖尿病组、糖尿病对照组和糖尿病给药组。通过免疫荧光方法验证巨噬细胞标记物F4/80和iba1在CX3CR1-GFP^(+)细胞中的表达;通过激光共聚焦显微镜观察CX3CR1-GFP^(+)细胞形态和数目分析结肠黏膜巨噬细胞的活化数量;通过RNAscope技术检测精氨酸酶1(arginase 1,Arg1)和一氧化氮合酶(inducible nitric oxide synthase,iNOS)的表达和分布变化。结果免疫荧光结果显示F4/80和iba1免疫阳性的细胞与CX3CR1-GFP^(+)的细胞基本共定位。与非糖尿病小鼠相比,STZ诱导的1型糖尿病模型小鼠结肠黏膜层活化的巨噬细胞数目增多(P<0.001),且M1型巨噬细胞产物iNOS在黏膜层腔侧表达增加(P<0.001),而M2型巨噬细胞产物Arg1在黏膜固有层表达下降(P<0.001);与糖尿病对照组相比,给予5-HT_(4)R激动剂可以抑制活化的巨噬细胞数目(P<0.001),结肠黏膜iNOS的表达上调(P<0.01)和Arg1的表达下调(P<0.05)。结论5-HT_(4)R激动剂可以抑制糖尿病小鼠结肠黏膜巨噬细胞向M1型极化。Objective To investigate the effects of 5-HT_(4)R agonist on the macrophage polarization in the colonic mucosa of diabetic mice.Methods Type 1 diabetes was induced by intraperitoneal injection of streptozotocin(STZ)in CX3CR1-GFP^(+)C57BL/6 mice.5-HT_(4)R agonist RS67333 was used for therapeutic intervention.The mice were randomly divided into control group,diabetic group(group D)and RS67333 intervention group(D-RS group).The expression of macrophage biomarkers F4/80 and ionized calcium binding adaptor molecule 1(iba1)in CX3CR1-GFP^(+)cells was detected by immuno-fluorescence.The activation of macrophages was assessed by observing the change in morphology and number of CX3CR1-GFP^(+)cells.The expression and distribution of arginase(Arg1)and inducible nitric oxide synthase(iNOS)in colon was detected by RNAscope.Results Immunofluorescent assay revealed that F4/80-or iba1-immunoreactive cells and CX3CR1-GFP^(+)cells were co-localized in cells.Compared with normal control group,the activated macrophages in diabetic group were significantly increased(P<0.001).The expression of iNOS,production of M1 macrophages was increased in the mucosa close to lumen(P<0.001),while the expression of Arg1,production of M2 macrophages was decreased in the laminae propria in diabetic mice(P<0.001).5-HT_(4)R agonist inhibited diabetes-induced increase of activated macrophages(P<0.001),upregulation of iNOS(P<0.01)and downregulation of Arg1(P<0.05)in the mucosa of colon.Conclusion 5-HT_(4)R agonist can inhibit the shift in macrophage polarization from M2 to M1 in diabetic mice.

关 键 词：糖尿病 巨噬细胞极化 5-羟色胺4受体 

分 类 号：R574[医药卫生—消化系统]