STZ诱导1型糖尿病小鼠早期胰腺巨噬细胞改变的实验研究  

Pancreatic macrophage changes in early stage of streptozotocin induced type 1 diabetes mellitus

作　　者：苏菲娅 刘伟江 张金霞 张晗[1,2] 于丰实 刘元林 李雪 王洋 郑荣秀 张毅[1,2] SU Fei-ya;LIU Wei-jiang;ZHANG Jin-xia;ZHANG Han;YU Feng-shi;LIU Yuan-lin;LI Xue;WANG Yang;ZHENG Rong-xiu;ZHANG Yi(Pediatric Department,Tianjin Medical University General Hospital,Tianjin 300052,China;Institute of Radiation Medicine,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China)

机构地区：[1]天津医科大学总医院儿科,天津300052 [2]军事科学院军事医学研究院辐射医学研究所,北京100850

出　　处：《军事医学》2021年第9期685-691,共7页Military Medical Sciences

基　　金：国家重点研发计划(2016YFC1000305);天津市卫计委科技基金攻关项目(16KG123);天津市科技委重点资助项目(17JCZDJ36400);天津医科大学总医院青年孵育基金(303073000501)。

摘　　要：目的探究链脲佐菌素(STZ)诱导1型糖尿病(T1DM)小鼠早期胰腺巨噬细胞的时空改变。方法36~42日龄C57BL/6J小鼠,连续5 d腹腔注射小剂量[50 mg/(kg·d)]STZ构建T1DM模型,观察小鼠的一般状态、体质量、血糖改变,HE染色观察胰岛病理学变化,免疫荧光法及免疫组化法检测胰岛素分泌情况。分别于STZ给药后第1、3、5、7天取小鼠胰腺,免疫荧光法检测胰岛巨噬细胞浸润,实时荧光定量PCR(RT-PCR)检测胰腺巨噬细胞相关炎症因子白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、一氧化氮合酶2(NOS_(2))的表达。结果STZ诱导T1DM小鼠出现典型的多饮、多食、多尿、体质量增长减缓、血糖明显升高的症状;病理学检测结果表明,T1DM组胰岛组织损伤严重,胰岛面积显著减小;免疫荧光及免疫组化结果显示,T1DM组较对照组胰岛素阳性细胞和胰岛素分泌明显减少,胰岛结构不完整,提示模型建立成功。进一步利用免疫荧光检测不同时间点巨噬细胞改变,结果表明,STZ给药后即开始出现巨噬细胞浸润增加,至第3天后明显增多,第5天达高峰,第7天出现下降趋势;相应胰岛的破坏出现在血糖增高或临床症状出现之前;RT-PCR检测相关炎症因子的表达,结果显示,IL-1β、TNF-α、NOS;的表达逐渐升高,至第5天达高峰后下降。结论STZ诱导T1DM小鼠模型早期即出现胰腺M1型巨噬细胞增多,其改变与胰腺炎症的发生呈时间依赖性。Objective To explore the temporal and spatial changes in pancreatic macrophages in the early stage of streptozotocin(STZ)induced type 1 diabetes mellitus(T1DM)mouse models. Methods T1DM was induced in C57BL/6J mice aged 36 to 42 days intraperitoneally injected with 50 mg/kg STZ for 5 days. The changes in the general physiological state,body mass,and blood glucose in T1DM mice were monitored. The pathological changes in islets and insulin secretion were examined by HE staining,immunofluorescence and immunohistochemistry,respectively. After STZ administration on days 1,3,5,and 7,the number of pancreatic macrophages was determined by immunofluorescence. Moreover,the expression levels of inflammatory factors related to pancreatic macrophage including interleukin 1β(IL-1β),tumor necrosis factor-α(TNF-α),and nitric oxide synthase 2(NOS_(2)) were detected by real-time fluorescence quantitative PCR(RT-PCR). Results Typical symptoms of diabetes,including polydipsia,polyphagia,polyuria,slowed weight growth,and a significant increase in blood glucose occurred in T1DM mice induced by STZ. Pathological examination results revealed serious islet tissue damage in mice in STZ group,and the islet area was significantly reduced. The results of immunofluorescence and immunohistochemistry showed that the insulin positive cells and insulin secretion significantly decreased,and the islet structure was incomplete,suggesting that the T1DM model was established successfully. The changes in macrophages at different time points were detected by immunofluorescence assay. The results determined that macrophage infiltration began to increase immediately after STZ injection,significantly increased on d 3,peaked on d5,and trended on down d 7. The islet destruction was presented before an increase in blood glucose or onset of clinical symptoms and matched the trend of macrophage infiltration. The expressions of related inflammatory cytokines were detected by RT-PCR,and the results showed that the expressions of IL-1β,TNF-α and NOS;increase

关 键 词：1型糖尿病 巨噬细胞 链脲佐菌素 胰腺 白细胞介素1Β 肿瘤坏死因子α 

分 类 号：R587.1[医药卫生—内分泌]