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利用小动物成像技术对小鼠胰岛β细胞容量在体非创评估
The in vivo non-invasive measurement of mouse pancreatic β cell mass through bioluminescence imaging technology
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DOI:
作者:
赵瑜,林紫薇,孙赟,慕开达,王琛*,贾伟平
ZHAO Yu, LIN Ziwei, SUN Yun, MU Kaida, WANG Chen*, JIA Weiping
作者单位:
上海交通大学附属第六人民医院内分泌代谢科,上海市糖尿病研究所,上海市糖尿病重点实验室
The Department of Endocrinology and Metabolism, Shanghai Jiao Tong University Affiliated Sixth People’s Hospital; Shanghai Diabetes Institute; Shanghai Key Laboratory of Diabetes Mellitus
关键词:
胰岛素抵抗 ; 糖尿病; β细胞 ; 小动物活体成像
Insulin resistance, Diabetes , β Cell mass, Bioluminescence imaging
摘要:
目的 利用β细胞中携带有荧光素酶报告基因的转基因(MIP-TF)小鼠,并通过活体成像系统对该小鼠生理及病理状态下的β细胞容量进行在体评估,从而建立基于该小鼠、对β细胞进行在体追踪的小动物活体成像平台。方法 MIP-TF小鼠从美国Jackson Lab购置。葡萄糖耐量实验与葡萄糖刺激的胰岛素分泌实验分别采用小鼠腹腔注射1.5g/kg, 3.0g/kg体重葡萄糖进行。45%高脂饮食(high fat diet, HFD)喂养MIP-TF小鼠制备饮食诱导的肥胖小鼠;200mg/kg 链脲佐菌素(stretozotoxin, STZ)腹腔注射制备小鼠糖尿病模型。胰腺胰岛素免疫荧光染色用于胰岛β细胞组织学检测,小动物活体成像检测MIP-TF小鼠胰岛β细胞容量。结果 MIP-TF转基因小鼠与同窝野生型小鼠糖耐量及胰岛素分泌水平无统计学差异;MIP-TF小鼠的活体成像中荧光信号位于小鼠胰腺区域,其荧光强度在小鼠注射葡萄糖后明显增加。MIP-TF小鼠在HFD喂养后体重增加,反映胰岛β细胞容量的荧光强度显著增强,是普食喂养小鼠的4倍;而小鼠注射STZ后活体成像荧光强度几乎完全消失。以上活体成像结果与胰岛素免疫荧光结果一致。结论 MIP-TF小鼠β细胞中特异性表达荧光素酶不影响小鼠发育及葡萄糖代谢。利用该小鼠进行活体胰岛β细胞容量检测特异性高、敏感性强。该小鼠可作为一种工具动物平台,应用于糖尿病模型中β细胞容量的动态检测。该项工作的开展为国内连续、动态的研究β细胞容量提供了新途径。
Objective To evaluate the platform for measurement of pancreatic β cell mass through transgenic mice carrying luciferase reporter gene (MIP-TF) specific in β cells and non-invasive bioluminescence imaging technique. Methods MIP-TF mice were purchased from Jackson Lab. Glucose tolerance tests (GTT) and glucose-stimulated insulin secretion tests were conducted through glucose (1.5g / kg for GTTs, 3.0g / kg for insulin secretion tests) injection intra-peritoneally. MIP-TF mice were fed with either a chow food or a high-fat diet (HFD) (45% kcal fat). Diabetic mice were produced by stretozotocin (200mg/kg) injection intraperitoneally. Immunofluorescence histology studies were conducted to detect insulin staining in pancreas. The pancreatic β cell mass was determined by non-invasive bioluminescence imaging. Results There were no significant differences in glucose tolerance and glucose stimulated insulin secretion levels between MIP-TF and WT mouse. The fluorescent signal were mainly located in pancreatic area in MIP-TF mice. The fluorescence intensity increased significantly after glucose injection. Pancreatic β cell mass represented by bioluminescence intensity was increased to four-fold higher in HFD MIP-TF mice than in normal chow control mice, while the fluorescence intensity disappeared completely after STZ injection, both of which were inconsistent with the findings from insulin immunofluorescence data. Conclusion β cell-specific expression of the luciferase does not affect the growth and glucose metabolism of MIP-TF mice. The platform based on MIP-TF mice for in vivo pancreatic β cells mass detection technology has high sensitivity and specificity, and can be used as an animal model tool for detecting β cell mass dynamically in diabetic researches.
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