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STZ 诱导糖尿病模型

链脲佐菌素 (STZ) 单次或多次小剂量诱导胰岛 β 细胞损伤。

Study design

小鼠经口灌胃葡萄糖负荷的耐受试验。禁食 4–6 小时；普通饮食按 1 g/kg、高脂饮食 4 周以上按 0.5 g/kg 给予 50% 葡萄糖溶液；负荷前及负荷后 15、30、60、90、120 分钟测定血糖。不同组别与样本量按具体研究设计确定，本方案不预设实验分组。

Outcome measures

以血糖浓度对时间作图得到完整血糖-时间曲线，并可计算曲线下面积（AUC）反映葡萄糖清除程度；当各受试对象基线血糖不同时应比较高于基线的幅度或曲线下面积，而非仅比较单一终点值。该检测反映葡萄糖负荷后的整体清除，不等同于胰岛素作用的直接测量。

基线与负荷后 15、30、60、90、120 分钟血糖时间序列；可计算血糖曲线下面积。血浆胰岛素结果按所用 ELISA 的校准与量程判读。

340 nm吸光度变化对应的葡萄糖浓度。

Experimental animals

血液葡萄糖浓度酶法测定
Tissue
blood glucose assay samples

Experimental procedures

提前一周单笼饲养 实验前一周将小鼠改为单笼饲养。 称量体重 逐只称量小鼠体重，用于按体重计算给糖体积。 禁食6小时 禁食六小时（06:00:00）。 测定基线血糖 禁食结束后，先用血糖仪测定血糖作为基线。 腹腔注射葡萄糖 按2 g/kg体重腹腔注射葡萄糖：将2 g D-葡萄糖溶于10 ml生理盐水配成20%葡萄糖贮备液，按每克体重10 ul给予。 系列采血测定血糖 于注射葡萄糖后15、30、45、60、90和120分钟采集血样，用血糖仪测定血糖。

禁食 Fasting begins first thing in the morning (around 9am). Fast mice for 4-6 hours before OGTT begins. Transfer mice to clean cage and wire top. Keep water bottles during the fasting period. 1. If using High Fat diet (HFD), save food to give back at the end of the OGTT. 1. mark tails with a sharpie for easier identification during OGTT. 口服葡萄糖负荷 Weigh mouse to obtain body weight for dose calculations. Calculate bolus dose of glucose (**1g/kg**). Weight of mouse (g) x 2 = ul sterile glucose (50% dextrose) solution *If using HFD for 4 weeks or more, use 0.5g/kg (weight of mouse (g) x 1 = ul sterile glucose (50% dextrose) solution) 口服葡萄糖负荷 Prepare syringes using 1cc syringe and 18G oral gavage needle. Load all syringes with calculated glucose dose and position in front of home cage. 口服葡萄糖负荷 Prior to the delivery of the sterile glucose (50% dextrose) solution, a time “zero” blood glucose level must be measured. Restrain mouse and clean tail with 70% ethanol/gauze. Extract a small amount of blood from the tail vein onto One Touch Ultra blood glucose strips used with the One Touch Ultra 2 blood glucose monitoring system. Blood can be collected into microvette tubes that can be used to extract plasma which can be assayed for plasma insulin (see step 9 for plasma collection). Microvette tubes are blood collection tubes with a capillary. Tap tube until all the blood is collected in the tube. Place on ice until plasma collection. 口服葡萄糖负荷 Administer the correct dose of sterile glucose (50% dextrose) solution using the syringe and gavage needle. - Restraint of the Mouse, avoiding an excessively tight grip of the mouse, as this will induce a panic response. Ensure that the mouse cannot bite you and cannot move its head. - Gravity alone should be used to move the cannula down the esophagus. While gliding the cannula, rotate slightly as the cannula goes past the epiglottis and into the esophagus.The mouse should swallow the cannula, and this can be observed. - Inject the fluid slowly. - The cannula is then gently removed and the animal is returned to its cage. 口服葡萄糖负荷 Begin timing from the point of successfully delivering the sterile glucose (50% dextrose) solution. 口服葡萄糖负荷 Test and record the blood glucose level (see step 4) at 15, 30, 60, 90 and 120 minutes after the initial glucose delivery. Record each value and the condition of the animal on the Glucose Tolerance Test record sheet. Extract a small amount of blood from the tail vein at 15, 30, 60 and 120 minutes for plasma; skip blood collection at 90 minutes to ensure not to exceed 0.007 ml blood/ g body weight. 口服葡萄糖负荷 Return food and environmental enrichments to cages. 血浆采集 Place tubes into pre-chilled centrifuge. Centrifguge for 2 mins at 2000 rpm. 00:02:00 4 °C 血浆采集 Remove the callipary-lid and dispose in biohazard bin. Place the base lid and seal the microvette (click position). 血浆采集 Centrifuge at 4°C for 10 mins at 10 000 rpm. 00:10:00 4 °C 血浆采集 Using a pipette, transfer the plasma (top transparent layer) to a clean and labelled tube. Store plasma samples at -20°C until assayed. 血浆胰岛素测定 Assay plasma insulin with the STELLUX Chemi Rodent Insulin ELISA kit (ALPCO #80-INSMR-CH01) according to the manufacturer's protocol. Include the kit standards and quality controls in each assay and read the plate by the kit's chemiluminescence procedure.

Calibrate Cobas for Glucose analysis by running a multi analyte standard and two control serum. Calibrate Cobas for Glucose analysis by running a multi analyte standard and two control serum. Sample Handling as performed by the Cobas Mira Plus. Sample Handling as performed by the Cobas Mira Plus. a) Pipette 3 µL of sample into cuvette. b) Absorbance is measured at 340 nm. c) Add 100 µL of Glucose liquid reagent. d) Mixture is incubated at 37ºC for 10 minutes. e) Absorbance is measured at 340 nm. Change in absorbance is calculated.

Model identity

Model name
STZ 诱导糖尿病模型
Species
mouse
Taxonomy ID
10090
Strain
C57BL/6J / BALB/c
Genetic background
化学诱导模型：模型由链脲佐菌素对胰岛 β 细胞的毒性暴露定义，不靠引入遗传突变形成。本条目宿主品系限于 C57BL/6J 与 BALB/c：C57BL/6J 有外部公开资源的已验证诱导方案可资参考；BALB/c 的 STZ 给药方案有系统比较研究，但该品系对 STZ 相对不敏感、成模不稳定。品系间对 STZ 效应存在包括遗传抗性在内的差异，造模条件须按品系与方案验证。外部公开验证记录与公司内部验证分列，本条目不含后者。

Biological characteristics

Phenotype stage
单次大剂量方案以急性 β 细胞损伤为主：STZ 经 GLUT2 摄入后作为 DNA 烷基化剂造成链断裂，PARP 驱动 NAD+ 与 ATP 耗竭，亚硝基脲基团释放 NO 损伤线粒体氧化代谢；多次小剂量（MLD-STZ）方案呈较渐进、自身免疫样的 β 细胞破坏，表型更接近 1 型糖尿病。某一外部验证服务流程以非空腹血糖超过 250 mg/dL 作为其成模确认阈值；该阈值为特定流程判据，不是所有 STZ 模型的固有统一标准。
Research questions
STZ 剂量与方案（单次大剂量对比多次小剂量）如何决定 β 细胞损伤模式、程度与血糖表型；品系差异（如 C57BL/6J 常用、BALB/c 相对耐受）对成模可靠性与模型适用性的影响；GLUT2 介导摄取与 DNA 烷基化/PARP 激活通路在 β 细胞选择性毒性中的作用。
Limitations
成模效果依赖品系、剂量与操作条件，且为化学损伤模型：其 β 细胞破坏不等同于自发遗传性自身免疫糖尿病的全部免疫遗传背景。GLUT2 在啮齿类 β 细胞的表达远高于以 GLUT1 为主的人 β 细胞，跨种属解读需谨慎；BALB/c 对 STZ 相对不敏感、成模不稳定，有研究结论认为其用于 1 型糖尿病移植流程研究并非有效模型。

Experimental methods

Experimental protocols

  • 小鼠腹腔注射葡萄糖耐量检测（IPGTT）

    Intraperitoneal glucose tolerance test (IPGTT) in mice: single-house for one week, weigh, fast for 6 hours, measure baseline glucose, inject 2 g/kg body weight of 20% D-glucose intraperitoneally, then collect blood at 15 to 120 minutes after injection. Used to assess glucose clearance in obesity and glucose-metabolism models.

  • 小鼠口服葡萄糖耐量试验（OGTT）

    This protocol adopts the protocols.io Oral Glucose Tolerance Test in Mouse v1. Mice are fasted for 4–6 hours, weighed, and given oral glucose at 1 g/kg (0.5 g/kg after at least 4 weeks of high-fat diet). Blood glucose is measured before and at 15, 30, 60, 90 and 120 minutes after the load, with tail-vein plasma collected at selected time points for insulin measurement. The record describes the assay procedure, not executed disease-model results.

  • 血液葡萄糖浓度酶法测定

    Enzymatic blood glucose quantification by coupled hexokinase and glucose-6-phosphate dehydrogenase reactions, using absorbance change at 340 nm on the Cobas Mira Plus.

  • 小鼠口服葡萄糖耐量试验（OGTT）

    This protocol adopts the protocols.io Oral Glucose Tolerance Test in Mouse v1. Mice are fasted for 4–6 hours, weighed, and given oral glucose at 1 g/kg (0.5 g/kg after at least 4 weeks of high-fat diet). Blood glucose is measured before and at 15, 30, 60, 90 and 120 minutes after the load, with tail-vein plasma collected at selected time points for insulin measurement. The record describes the assay procedure, not executed disease-model results.

  • 小鼠腹腔注射葡萄糖耐量检测（IPGTT）

    Intraperitoneal glucose tolerance test (IPGTT) in mice: single-house for one week, weigh, fast for 6 hours, measure baseline glucose, inject 2 g/kg body weight of 20% D-glucose intraperitoneally, then collect blood at 15 to 120 minutes after injection. Used to assess glucose clearance in obesity and glucose-metabolism models.

  • 小鼠口服葡萄糖耐量试验（OGTT）

    This protocol adopts the protocols.io Oral Glucose Tolerance Test in Mouse v1. Mice are fasted for 4–6 hours, weighed, and given oral glucose at 1 g/kg (0.5 g/kg after at least 4 weeks of high-fat diet). Blood glucose is measured before and at 15, 30, 60, 90 and 120 minutes after the load, with tail-vein plasma collected at selected time points for insulin measurement. The record describes the assay procedure, not executed disease-model results.

  • 小鼠腹腔注射葡萄糖耐量检测（IPGTT）

    Intraperitoneal glucose tolerance test (IPGTT) in mice: single-house for one week, weigh, fast for 6 hours, measure baseline glucose, inject 2 g/kg body weight of 20% D-glucose intraperitoneally, then collect blood at 15 to 120 minutes after injection. Used to assess glucose clearance in obesity and glucose-metabolism models.

  • 小鼠口服葡萄糖耐量试验（OGTT）

    This protocol adopts the protocols.io Oral Glucose Tolerance Test in Mouse v1. Mice are fasted for 4–6 hours, weighed, and given oral glucose at 1 g/kg (0.5 g/kg after at least 4 weeks of high-fat diet). Blood glucose is measured before and at 15, 30, 60, 90 and 120 minutes after the load, with tail-vein plasma collected at selected time points for insulin measurement. The record describes the assay procedure, not executed disease-model results.

  • 小鼠口服葡萄糖耐量试验（OGTT）

    This protocol adopts the protocols.io Oral Glucose Tolerance Test in Mouse v1. Mice are fasted for 4–6 hours, weighed, and given oral glucose at 1 g/kg (0.5 g/kg after at least 4 weeks of high-fat diet). Blood glucose is measured before and at 15, 30, 60, 90 and 120 minutes after the load, with tail-vein plasma collected at selected time points for insulin measurement. The record describes the assay procedure, not executed disease-model results.

  • 小鼠口服葡萄糖耐量试验（OGTT）

    This protocol adopts the protocols.io Oral Glucose Tolerance Test in Mouse v1. Mice are fasted for 4–6 hours, weighed, and given oral glucose at 1 g/kg (0.5 g/kg after at least 4 weeks of high-fat diet). Blood glucose is measured before and at 15, 30, 60, 90 and 120 minutes after the load, with tail-vein plasma collected at selected time points for insulin measurement. The record describes the assay procedure, not executed disease-model results.

  • 小鼠口服葡萄糖耐量试验（OGTT）

    This protocol adopts the protocols.io Oral Glucose Tolerance Test in Mouse v1. Mice are fasted for 4–6 hours, weighed, and given oral glucose at 1 g/kg (0.5 g/kg after at least 4 weeks of high-fat diet). Blood glucose is measured before and at 15, 30, 60, 90 and 120 minutes after the load, with tail-vein plasma collected at selected time points for insulin measurement. The record describes the assay procedure, not executed disease-model results.

  • 小鼠腹腔注射葡萄糖耐量检测（IPGTT）

    Intraperitoneal glucose tolerance test (IPGTT) in mice: single-house for one week, weigh, fast for 6 hours, measure baseline glucose, inject 2 g/kg body weight of 20% D-glucose intraperitoneally, then collect blood at 15 to 120 minutes after injection. Used to assess glucose clearance in obesity and glucose-metabolism models.

Reagents and materials

References

  1. Like AA, Rossini AA. Streptozotocin-induced pancreatic insulitis: new model of diabetes mellitus. Science. 1976;193(4251):415-417. DOI:10.1126/science.180605; PMID:180605. 摘要记载 STZ 在小鼠（mice）诱发胰腺胰岛炎，未注明具体品系。
  2. Dewalkar LP. Experimental models in diabetes research. Lab Anim Res. 2026;42(1):25. DOI:10.1186/s42826-026-00286-6; PMID:42387633; PMCID:PMC13326422. 正文记载：STZ 为经 GLUT2 进入 β 细胞的糖胺-亚硝基脲类化合物，主经 DNA 烷基化链断裂、PARP 驱动的 NAD+ 与 ATP 耗竭及 NO 释放致线粒体氧化代谢障碍杀伤 β 细胞；单次大剂量与多次小剂量（MLD-STZ）渐进自身免疫样破坏两种方案；GLUT2 在啮齿类远高于以 GLUT1 为主的人细胞。正文未把 C57BL/6J 或 BALB/c 列为 STZ 品系。
  3. The Jackson Laboratory. STZ-Induced Diabetes service page: "Diabetes is induced in C57BL/6J, NOD-scid, and NSG male mice using validated protocols"; "only provide mice with non-fasting blood glucose exceeding 250 mg/dL"；STZ described as an alkylating agent targeting insulin-producing beta cells, mimicking Type 1 diabetes phenotypes in mice. 访问于 2026-10-18；为外部公开资源的服务流程判据，不构成吉田内部验证或供应承诺。
  4. Wszola M, Klak M, Kosowska A, Tymicki G, Berman A, Adamiok-Ostrowska A, et al. Streptozotocin-Induced Diabetes in a Mouse Model (BALB/c) Is Not an Effective Model for Research on Transplantation Procedures in the Treatment of Type 1 Diabetes. Biomedicines. 2021;9(12):1790. DOI:10.3390/biomedicines9121790; PMID:34944607; PMCID:PMC8698562. 研究直接使用 40 只 BALB/c 小鼠行 STZ 给药并比较八种方案；讨论记载品系间对 STZ 存在包括遗传抗性在内的差异；结论认为 BALB/c 用于该类移植流程研究并非有效模型。
  5. NCBI Taxonomy record 10090: Mus musculus (house mouse), rank species.
  6. Roberta Marongiu, Sabina Marciano. Glucose Tolerance Test, Version 2. protocols.io record 98408, 2024-04-18. https://doi.org/10.17504/protocols.io.261geddodv47/v2
  7. Standard operating procedures for describing and performing metabolic tests of glucose homeostasis in mice. Disease Models & Mechanism (2010), PMID 20713647, DOI 10.1242/dmm.006239
  8. Nancy Smith, Mourad Ferdaoussi, Haopeng Lin, Patrick Macdonald. Oral Glucose Tolerance Test in Mouse. protocols.io. https://doi.org/10.17504/protocols.io.ujjeukn
  9. Benedé-Ubieto R, Estévez-Vázquez O, Ramadori P, Cubero FJ, Nevzorova YA. Guidelines and Considerations for Metabolic Tolerance Tests in Mice. Diabetes Metab Syndr Obes. 2020;13:439-450. PMID 32110077. doi:10.2147/DMSO.S234665
  10. ALPCO. STELLUX Chemi Rodent Insulin ELISA (Catalog #80-INSMR-CH01) instructions.
  11. protocols.io · vv1 · John Stack; Gary Cline · This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
  12. ARRIVE 2.0 guidelines
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