Western or fast food style diets fed to induce NASH with metabolic syndrome contain 40 – 45% kcal from milkfat (a fat source high in palmitate) with added cholesterol (0.15 – 2%) and are high in sucrose (>30%). Dietary palmitate and cholesterol have both previously been associated with the progression from simple steatosis to NASH.

Examples:

• TD.88137       Adjusted Calories Diet (42% from fat)
• TD.96121       21% MF, 1.25% Chol. Diet
• TD.120528     42% Kcal/Fat Diet (Incr. Sucrose, 1.25% Chol.)

Research use:

These diets can induce obesity, metabolic syndrome, and simple steatosis within nine weeks of feeding. Increased hepatic inflammation has been observed after 12 weeks of feeding. NASH typically requires longer feeding with fibrosis developing within nine months and late stage fibrosis including hepatic ballooning occurring after 14 – 20 months of feeding. Increasing dietary sucrose (~41%) and cholesterol (~1.25%) accelerates the NASH phenotype with steatosis, inflammation and hepatocyte ballooning observed within 12 weeks. In addition to feeding a high fat diet, providing a glucose/fructose mixture in the drinking water may further promote NASH development.

Select References:

Charlton, M., et al., Fast food diet mouse: novel small animal model of NASH with ballooning, progressive fibrosis, and high physiological fidelity to the human condition. Am J Physiol Gastrointest Liver Physiol, 2011. 301(5): p. G825-34. http://www.ncbi.nlm.nih.gov/pubmed/21836057

Gores, G., Charlton M, Krishnan A, Viker K, Sanderson S, Cazanave S, McConico A, Masuoko H. Am J Physiol Gastrointest Liver Physiol, 2015. 308: p. G159. http://ajpgi.physiology.org/content/308/2/G159

Li, Z.Z., et al., Hepatic lipid partitioning and liver damage in nonalcoholic fatty liver disease: role of stearoyl-CoA desaturase. J Biol Chem, 2009. 284(9): p. 5637-44. http://www.ncbi.nlm.nih.gov/pubmed/19119140

Ioannou, G.N., et al., Hepatic cholesterol crystals and crown-like structures distinguish NASH from simple steatosis. J Lipid Res, 2009. 54(5): p. 1326-34. http://www.ncbi.nlm.nih.gov/pubmed/23417738

Alkhouri, N., et al., Adipocyte apoptosis, a link between obesity, insulin resistance, and hepatic steatosis. J Biol Chem, 2010. 285(5): p. 3428-38. http://www.ncbi.nlm.nih.gov/pubmed/19940134

Dixon, L.J., et al., Caspase-1 as a central regulator of high fat diet-induced non-alcoholic steatohepatitis. PLoS One, 2013. 8(2): p. e56100. http://www.ncbi.nlm.nih.gov/pubmed/23409132

DeLeve, L.D., et al., Prevention of hepatic fibrosis in a murine model of metabolic syndrome with nonalcoholic steatohepatitis. Am J Pathol, 2008. 173(4): p. 993-1001. http://www.ncbi.nlm.nih.gov/pubmed/18772330

VanSaun, M.N., et al., High fat diet induced hepatic steatosis establishes a permissive microenvironment for colorectal metastases and promotes primary dysplasia in a murine model. Am J Pathol, 2009. 175(1): p. 355-64. http://www.ncbi.nlm.nih.gov/pubmed/19541928

Asgharpour, A., et al., A diet-induced animal model of non-alcoholic fatty liver disease and hepatocellular cancer. J Hepatol, 2016. 65(3): p. 579-88. http://www.ncbi.nlm.nih.gov/pubmed/27261415

Tetri, L.H., et al., Severe NAFLD with hepatic necroinflammatory changes in mice fed trans fats and a high-fructose corn syrup equivalent. Am J Physiol Gastrointest Liver Physiol, 2008. 295(5): p. G987-95. http://www.ncbi.nlm.nih.gov/pubmed/18772365

Tsuchida, T., et al., A simple diet-and chemical-induced murine NASH model with rapid progression of steatohepatitis, fibrosis and liver cancer. Journal of hepatology, 2018. 69(2):385-395. https://www.ncbi.nlm.nih.gov/pubmed/29572095

The American Lifestyle-Induced Obesity Syndrome (ALIOS) model involves feeding the “American fast food” diet high in trans-fats and sugar. Dietary trans-fats from hydrogenated vegetable shortening (HVO) are associated with increased insulin resistance and hepatic inflammation in rodent NASH models. In addition to diet, a glucose/fructose solution is added to the drinking water and sedentary behavior promoted by removing the overhead cage feeders in this model.

Examples:

• TD.06303       22% HVO Diet
• TD.120330     22% HVO + 0.2% Cholesterol Diet
• TD.130885     ALIOS with Added Sugar

Research use:

The ALIOS model develops obesity with insulin resistance, elevated ALT levels, and steatosis within 16 weeks. Increased inflammation and early development of fibrosis have been observed at 6 months. Severe steatosis with fibrosis and inflammation develops within 12 months of feeding with 50% of the mice reportedly developing hepatic neoplasms. Adding cholesterol (0.2%) to the American Fast Food diet may accelerate NASH phenotype development.

Select References:

Koppe, S.W., et al., Trans fat feeding results in higher serum alanine aminotransferase and increased insulin resistance compared with a standard murine high-fat diet. Am J Physiol Gastrointest Liver Physiol, 2009. 297(2): p. G378-84. http://www.ncbi.nlm.nih.gov/pubmed/19541924

Tetri, L.H., et al., Severe NAFLD with hepatic necroinflammatory changes in mice fed trans fats and a high-fructose corn syrup equivalent. Am J Physiol Gastrointest Liver Physiol, 2008. 295(5): p. G987-95. http://www.ncbi.nlm.nih.gov/pubmed/18772365

Mells, J.E., et al., Glp-1 analog, liraglutide, ameliorates hepatic steatosis and cardiac hypertrophy in C57BL/6J mice fed a Western diet. Am J Physiol Gastrointest Liver Physiol, 2012. 302(2): p. G225-35. http://www.ncbi.nlm.nih.gov/pubmed/22038829

Dowman, J.K, et al., Development of hepatocellular carcinoma in a murine model of nonalcoholic steatohepatitis induced by use of a high-fat/fructose diet and sedentary lifestyle. Am J Pathol, 2014. 184(5):1550-1561. https://www.ncbi.nlm.nih.gov/pubmed/24650559 

Mells, J.E., et al., Saturated fat and cholesterol are critical to inducing murine metabolic syndrome with robust nonalcoholic steatohepatitis. J Nutr Biochem, 2014. 26(3): p. 285-92. http://www.ncbi.nlm.nih.gov/pubmed/25577467

The Fructose, Palmitate, Cholesterol and Trans-Fat (FPC) diet is a recent NASH diet that includes Western and ALIOS model diets to achieve both metabolic and hepatic NASH features within an accelerated time frame. Key features of the FPC diet include 1) a lower Met content than typical rodent diets by decreasing total protein without supplementing sulfur amino acids; 2) choline supplementation is lower than typical but is not considered deficient; 3) high in sucrose (~34% by weight); 4) 1.25% cholesterol; 5) 52% kcal from fat with fat sources including milkfat fat, palmitic acid and hydrogenated vegetable shortening to provide trans-fats. Like the ALIOS model, the FPC model also provides a glucose/fructose solution to the drinking water.

Examples:

• TD.160785     52 kcal/Fat Diet (C16:0, HVO, AMF, Choline/Met)

Research use:

Male C57BL/6J mice fed the FPC diet and provided a glucose/fructose drinking solution developed insulin resistance and NAFLD with inflammation, hepatocyte death, and fibrosis within 16 weeks.

Select References:

Wang, X., et al., Hepatocyte TAZ/WWTR1 promotes inflammation and fibrosis in nonalcoholic steatohepatitis. Cell Metab, 2016. 24(6): p. 848-62. https://www.ncbi.nlm.nih.gov/pubmed/28068223  

Zhu, C., et al., Hepatocyte Notch activation induces liver fibrosis in nonalcoholic steatohepatitis. Sci Transl Med, 2018. 10(468). https://www.ncbi.nlm.nih.gov/pubmed/30463916

Common diets to induce obesity (DIO) can be fed to induce uncomplicated NAFLD. These high fat diets typically contain 40–60% kcal from fat without supplemented cholesterol or cholate. Simple sugars such as sucrose or fructose can also be supplemented via diet or water to progress the fatty liver phenotype. Diets can be in pellet or powder/dough form depending on the formula. Some models require limited physical activity and in those cases diets can be fed inside the cage. For more information see our Diet Induced Obesity page.

Examples:

• TD.08811       45%kcal Fat Diet (21% MF, 2% SBO)
• TD.06414       Adjusted Calories Diet (60/Fat)

Research use:

In susceptible rodent models, high fat diets are commonly used to induce NAFLD with obesity and insulin resistance common metabolic features associated with NASH in humans. However, the degree of NASH pathology (steatosis, inflammation, and fibrosis) is limited or mild and varies depending on the animal model, length of feeding, and dietary components.

Originally formulated to induce mild atherosclerosis in wild-type rodents, high fat diets containing added cholesterol (1 – 1.25%) and cholate (0.5% as sodium cholate or cholic acid) have also been useful in inducing NASH. This diet option includes purified “Western” style diets with increased cholesterol and cholate and also hybrid diets. Hybrid diets were originally developed by Beverly Paigen and colleagues by mixing a natural ingredient mouse diet in a 3:1 ratio with a concentrated purified diet (containing 5% cholesterol and 2% sodium cholate) resulting in a diet containing ~15.8% fat, 1.25% cholesterol, and 0.5% sodium cholate. Although a less refined approach, the hybrid diet is associated with increased gallstone formation and liver damage as compared to similar purified diets.

Examples:

• TD.02028       Atherogenic Rodent Diet
• TD.88051       Cocoa Butter Diet and Purina Mouse Chow
• TD.09237       15% AMF Diet (1% Chol, 0.5% NaChol)

Research use:

Atherogenic diets are able to induce varied degrees of NASH with increased hepatic inflammation with early fibrosis observed after ten weeks of feeding. However, the metabolic profile typical in human NASH (obesity with insulin resistance) is not recapitulated in this model with animals typically maintaining similar body weights as control fed groups without the development of metabolic syndrome.

Select References:

Nishina, P.M., J. Verstuyft, and B. Paigen, Synthetic low and high fat diets for the study of atherosclerosis in the mouse. J Lipid Res, 1990. 31(5): p. 859-69. http://www.ncbi.nlm.nih.gov/pubmed/2380634

Kamari, Y., et al., Lack of interleukin-1alpha or interleukin-1beta inhibits transformation of steatosis to steatohepatitis and liver fibrosis in hypercholesterolemic mice. J Hepatol, 2011. 55(5): p. 1086-94. http://www.ncbi.nlm.nih.gov/pubmed/21354232

Kim, D.G., et al., Non-alcoholic fatty liver disease induces signs of Alzheimer’s disease (AD) in wild-type mice and accelerates pathological signs of AD in an AD model. J Neuroinflammation, 2016. 13: p. 1. http://www.ncbi.nlm.nih.gov/pubmed/26728181

Madrigal-Perez, V.M., et al., Preclinical analysis of nonsteroidal anti-inflammatory drug usefulness for the simultaneous prevention of steatohepatitis, atherosclerosis and hyperlipidemia. Int J Clin Exp Med, 2015. 8(12): p. 22477-83. http://www.ncbi.nlm.nih.gov/pubmed/26885230

Savransky, V., et al., Chronic intermittent hypoxia causes hepatitis in a mouse model of diet-induced fatty liver. Am J Physiol Gastrointest Liver Physiol, 2007. 293(4): p. G871-7. http://www.ncbi.nlm.nih.gov/pubmed/17690174

Methionine and choline deficient (MCD) diets are amino acid defined rodent diets deficient in methionine and choline, high in sucrose (>40% by weight) with ~10% corn oil by weight. Methionine and choline deficiency decreases fat oxidation and export of fat from the liver. Dietary sucrose is necessary for hepatic lipid accumulation and oxidation. The polyunsaturated fat in corn oil promotes hepatic lipid oxidation.

Example:

• TD.90262       Methionine/Choline Deficient Diet

Control:

• TD.94149       Amino Acid Control Diet

Research use:

Steatosis, increased serum alanine aminotransferase (ALT), inflammation, and hepatic fat oxidation has been observed within three weeks of feeding the MCD diet with fibrosis development after six weeks. This dietary model does not produce metabolic syndrome (an aspect of NASH in human models) and progressive weight loss (up to 40%) is associated with the MCD diet feeding.

蛋氨酸/胆碱缺乏症(MCD)日粮
蛋氨酸和胆碱缺乏(MCD)饲料是一种氨基酸定义的啮齿动物饲料，缺乏蛋氨酸和胆碱，蔗糖含量高(体重>40%)，玉米油含量约10%。蛋氨酸和胆碱缺乏减少脂肪氧化和从肝脏输出脂肪。日粮蔗糖对肝脏脂质的积累和氧化是必需的。玉米油中的多不饱和脂肪促进肝脏脂质氧化。

例子：TD.90262 蛋氨酸/胆碱缺乏症饮食
管制：TD.94149 氨基酸控制饮食

研究用途：

观察到脂肪变性、血清丙氨酸转氨酶(ALT)升高、炎症和肝脂肪氧化反应。这种饮食模式不会产生代谢综合症(在人类模型中是NASH的一个方面)和累进性减肥(高达40%)与MCD饮食喂养有关。

Select References:

Pickens, M.K., et al., Dietary sucrose is essential to the development of liver injury in the MCD model of steatohepatitis. J Lipid Res, 2009. 50(10):2072-82.  http://www.ncbi.nlm.nih.gov/pubmed/19295183

Li, Z.Z., et al., Hepatic lipid partitioning and liver damage in nonalcoholic fatty liver disease: role of stearoyl-CoA desaturase. J Biol Chem, 2009. 284(9): p. 5637-44. http://www.ncbi.nlm.nih.gov/pubmed/19119140

Lee, G.S., et al., Polyunsaturated fat in the methionine-choline-deficient diet influences hepatic inflammation but not hepatocellular injury. J Lipid Res, 2007. 48(8): p. 1885-96. http://www.ncbi.nlm.nih.gov/pubmed/17526933

Vetelainen, R., A. van Vliet, and T.M. van Gulik, Essential pathogenic and metabolic differences in steatosis induced by choline or methione-choline deficient diets in a rat model. J Gastroenterol Hepatol, 2007. 22(9): p. 1526-33. http://www.ncbi.nlm.nih.gov/pubmed/17716355

Leclercq, I.A., et al., Intrahepatic insulin resistance in a murine model of steatohepatitis: effect of PPARgamma agonist pioglitazone. Lab Invest, 2007. 87(1): p. 56-65. http://www.ncbi.nlm.nih.gov/pubmed/17075577

Kashireddy, P.R. and M.S. Rao, Sex differences in choline-deficient diet-induced steatohepatitis in mice. Exp Biol Med (Maywood), 2004. 229(2): p. 158-62. http://www.ncbi.nlm.nih.gov/pubmed/14734794

Dixon, L.J., et al., Caspase-1-mediated regulation of fibrogenesis in diet-induced steatohepatitis. Lab Invest, 2012. 92(5): p. 713-23. http://www.ncbi.nlm.nih.gov/pubmed/22411067

Dietary models of NAFLD/NASH continue to evolve with the goal of more accurately recapitulating both the metabolic and hepatic symptoms of human disease. Commonly researchers are studying the synergistic effects of various NASH dietary features to accelerate progression of the model and severity of liver disease.

A Teklad nutritionist can work with you to formulate new diets in order to investigate novel dietary models of NAFLD/NASH.

The choice of control diet is dependent on the specific research goal. Many researchers choose to compare their NAFLD/NASH diet-fed animals to animals fed a natural ingredient, grain-based diet (also referred to as standard diet or chow). These diets differ in the source and level of nutrients as well as in the presence of non-nutritive factors (such as phytates or phytoestrogens).

Depending on what your main comparisons are, it may be suitable to have a grain-based diet as your control/reference group. However, making such comparisons limits inferences to dietary patterns versus a specific dietary component. In some cases, such as those studies feeding amino acid defined diets like the MCD model, a matched control diet is recommended given the very different formulations and protein sources of grain-based diets.

When making inferences about specific nutrients within the diet an ingredient matched, low fat control diet may be necessary. There are many options with different levels and types of fat in addition to different types of carbohydrate ranging from sucrose (highly refined and digestible) to corn starch (refined, but more complex) to resistant starch (refined, but not fully digestible).

A very basic purified control diet would be AIN-93M TD.94048 or AIN-93G TD.94045. AIN-93 diets have a moderate amount of sucrose at ~10% with fat from soybean oil providing a healthy fatty acid profile.

Contact a nutritionist for an additional information and control diet recommendations.

对照日粮
控制饮食的选择取决于具体的研究目标。许多研究人员选择比较他们的NAFLD/纳什饮食喂养的动物和喂养一种天然成分的动物，谷物为基础的饮食(也称为标准饮食或周食)。这些饮食在营养来源和水平以及非营养因素(如植酸盐或植物雌激素)存在的情况下存在差异。

根据您的主要比较，它可能适合作为您的对照/参考组谷物基础的饮食。然而，这样的比较限制了对饮食模式和特定饮食成分的推断。在某些情况下，如那些研究喂食氨基酸定义的饮食，如mcd模型，建议一个匹配的对照饮食，考虑到非常不同的配方和蛋白质来源的谷物为基础的饮食。

当对饮食中的特定营养成分做出相应的推断时，低脂控制饮食可能是必要的。除了不同类型的碳水化合物外，还有许多不同水平和类型的脂肪，从蔗糖(高精制和可消化)到玉米淀粉(精制，但更复杂)，再到抗性淀粉(精制，但不能完全消化)。

一种非常基本的纯正控制饮食将是-9300万。TD.94048或者是93g TD.94045。AIN-93日粮中含有适量的蔗糖~10%，大豆油中的脂肪提供了健康的脂肪酸谱。