High-fat diet-induced NASH Sprague-Dawley rat
Male Sprague-Dawley rats fed a high-fat diet containing approximately 60 to 71 kcal percent fat develop hepatocellular steatosis with hepatocyte ballooning injury and elevated hepatic triglycerides over 6 to 12 weeks. When combined with a single low-dose streptozotocin injection or initiated by diethylnitrosamine, the same diet drives more severe NASH-like inflammation and accelerates hepatocarcinogenesis, with NAS typically in the 5 to 7 range under methionine-choline-deficient regimens and lower NAS under high fat alone.
Outcome measures
血糖浓度读数。
获得可供光学显微镜观察的 H&E 染色组织切片:细胞核蓝紫色、细胞质与结缔组织纤维粉红色,用于观察组织中细胞的整体结构与形态细节。本方案本身不产出病理评分数值,也不提供特定分子标志物的定量结果。
Experimental animals
- Age
- as reported in cited studies
- Tissue
- blood
- Age
- as reported in cited studies
- Tissue
- liver
Experimental procedures
**Measurement of Blood Glucose ** **Measurement of Blood Glucose ** Blood samples were collected from the tail vein for the determination of blood glucose levels using Accutrend Plus test strips and meter (Roche).
脱蜡(Dewaxing) 石蜡切片置于二甲苯中脱蜡 3 次,每次 00:03:00。 复水(Rehydration) 石蜡切片依次置于 100% 乙醇中 3 次、每次 00:03:00,再置于 95% 乙醇中 1 次、00:03:00,随后用水漂洗。 苏木素核染色(Hematoxylin staining) 石蜡切片以苏木素染液染色 00:00:20,随后在流水下冲洗 00:01:00。 分化与返蓝(Differentiation and bluing) 石蜡切片置于返蓝液中 00:01:00,随后在流水下冲洗 00:01:00;此步使细胞核呈现蓝色。 伊红染色(Eosin staining) 石蜡切片以伊红复染 00:00:10;此步使细胞质成分与结缔组织纤维呈现粉红色。 脱水与透明(Dehydration and Clearing) 石蜡切片依次置于 95% 乙醇中 00:02:00、100% 乙醇中 2 次、每次 00:02:00,随后置于二甲苯中 2 次、每次 00:02:00。 封片(Mounting) 石蜡切片以 Permount 封片介质加盖玻片封固。
Model identity
- Model name
- High-fat diet-induced NASH Sprague-Dawley rat
- Species
- rat
- Taxonomy ID
- 10116
- Strain
- Sprague-Dawley
- Genetic background
- Sprague-Dawley
Biological characteristics
- Phenotype stage
- Male Sprague-Dawley rats, typically 8 weeks old and 180 to 200 g at start, are fed the high-fat diet for 6 to 12 weeks; diethylnitrosamine-initiated carcinogenesis protocols add a single priming injection followed by 6 weeks of high-fat feeding.
- Research questions
- The model has been used to evaluate hepatoprotective natural products such as lycopene, tomato extract and Dendrobium officinale polysaccharide against high-fat diet-induced MAFLD, and to compare NAFL, NASH and NAFL-plus-type-2-diabetes phenotypes across high-fat, methionine-choline-deficient and high-fat-plus-streptozotocin regimens in Sprague-Dawley rats.
- Limitations
- High-fat diet alone in Sprague-Dawley rats produces steatosis with modest NAS but limited progressive fibrosis over 12 weeks, so claims of advanced NASH require combination with methionine-choline deficiency or a carcinogen primer. The corpus does not document long-term fibrosis or cirrhosis endpoints, and diet compositions vary between studies, which limits cross-study comparison.
Experimental methods
Experimental protocols
- 大鼠尾静脉血糖测定
Measure rat tail-vein blood glucose using Roche Accutrend Plus test strips and meter.
- Hematoxylin and Eosin (H&E) staining for histopathology
Hematoxylin and eosin (H&E) staining of fixed, paraffin-embedded tissue sections (4 μm). The workflow covers dewaxing, rehydration, hematoxylin nuclear staining, bluing, eosin counterstaining, dehydration and clearing, and coverslipping, yielding blue-purple nuclei and pink cytoplasm and connective tissue fibers for light-microscopic assessment of cellular and tissue morphology.
Reagents and materials
- Accutrend Plus 血糖试纸与读数仪
血糖检测耗材与读数平台
- 组织学二甲苯(混合异构体)
脱蜡剂
- Ethanol
复水梯度溶剂(去除二甲苯)
- 95% 乙醇溶液
复水梯度溶剂
- 自来水(组织学染色用)
复水后漂洗用水(后续各流水冲洗共用)
- Epredia Signature Series 苏木素染液(7221)
细胞核染色
- Epredia Signature Series 返蓝液(7301)
返蓝(使细胞核转蓝)
- Epredia Signature Series 伊红 Y 染液(7111)
细胞质与结缔组织纤维复染
- Permount 组织学封片剂(Fisher SP15)
封片介质
References
- Dietary lycopene and tomato extract supplementations inhibit nonalcoholic steatohepatitis-promoted hepatocarcinogenesis in rats. Wang Yan; Ausman Lynne M; Greenberg Andrew S; et al. International journal of cancer 2010.
- Gene-metabolite network analysis in different nonalcoholic fatty liver disease phenotypes. Liu Xiao-Lin; Ming Ya-Nan; Zhang Jing-Yi; et al. Experimental & molecular medicine 2017.
- Hepatoprotective action of Dendrobium officinale polysaccharide against MAFLD: modulation of gut microbiota-metabolite cascade controls TLR4/NF-κB inflammatory response. Xu Yumei; Zhang Yanmei; Li Shuqing; et al. Frontiers in pharmacology 2026.
- protocols.io · vv2 · Michael Hill · 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
- Nathalia L. Oliveira; Samantha Gruenheid. Hematoxylin and Eosin (H&E) staining for histopathology. protocols.io, v1, 2026-07-31. DOI: 10.17504/protocols.io.kxygxr9jdg8j
- Epredia Signature Series Hematoxylin (7221), 产品信息页, Fisher Scientific
- Epredia Signature Series Bluing Reagent (7301), 产品信息页, Fisher Scientific
- Epredia Signature Series Eosin Y (7111), 产品信息页, Fisher Scientific
- Xylene, Fisherbrand HistoPrep (HC7001GAL), 产品信息页, Fisher Scientific
- Fisher Chemical Permount Mounting Medium (SP15 产品族), 产品信息页, Fisher Scientific
- Ethanol (CID 702, CAS 64-17-5), PubChem 化合物标准记录
- ARRIVE 2.0 guidelines