Name: rapid model to evaluate the anti-obesity potential of a combination of syzygium aromaticum (clove) and cuminun cyminum (cumin) on c57bl6/j mice fed high-fat diet
Uploaded: 2021-07-31T13:00:00.000+00:00
Duration: 13 min 9 s
Description: Watch this Scientific Journal Video about Rapid Model to Evaluate the Anti-Obesity Potential of a Combination of Syzygium aromaticum Clove and Cuminun cyminum Cumin on C57BL6/j Mice Fed High-Fat Diet

This research was supported by Instituto Polit&#233;cnico Nacional M&#233;xico.

All animal experiments were approved by the Animal Experiment Committee of Escuela Nacional de Ciencias Biologicas (IPN; Protocol No. 2732).
1. Preparation of the extract of the combination of cloves and cumin
NOTE: The edible plants, Syzygium aromaticum (clove) and Cuminum cyminum (cumin) were purchased from Central de Abastos CDMX, Mexico.
<ol>
	<li>Grind 500 g of seeds from the plants and perform ultrasonic-assisted extraction. Sonicate them (320 Watts; 24 kHz frequency, 30 min) with ethanol:water (60:40 v/v) at a temperature range of 30 &#177; 4 &#176;C.13</li>
	<li>Filter the solids using Whatman number 2 filter paper under vacuum and concentrate the extract with a rotary evaporator.</li>
	<li>Measure the UV-vis spectrum of the extract of both plants (CC).</li>
</ol>
2. Animals
<ol>
	<li>Use male C57BL6/J mice of six weeks old (48 in total, body weight 24-29 g) for the study.</li>
	<li>House the animals in groups of eight in standard laboratory conditions: temperature (20-22 &#177; 1 &#176;C), relative humidity (45-54 &#177; 2%), lighting (08:00-20:00 h) with food and water ad libitum. Acclimate the mice in these conditions for a week before the experiment.</li>
</ol>
3. Experimental Procedure
<ol>
	<li>Prepare the high fat diet14 as in Table 1.</li>
	<li>At the beginning of the test, subject each of the groups formed (n = 8) to different treatments: Feed Group 1 on a normal diet; Group 2, on high-fat diet; Group 3, on high-fat diet + 100 mg/kg of CC extract; Group 4, on high-fat diet + 250 mg/kg of CC extract; Group 5, on high-fat diet + 450 mg/kg of CC extract; Group 6, on high-fat diet + phentermine (an antiobesity drug used as positive control)15.</li>
	<li>Feed all the groups of mice for 5 weeks and record the amount of food consumed daily.
	<ol>
		<li>In a test tube with stopper, homogenize the extract or the drug with the dose of water corresponding to the group to which the treatment is being administered. This is done with vortex shaker.</li>
		<li>Once homogenized, add all the corresponding treatment into the drinker of each group.</li>
	</ol>
	</li>
</ol>
4. Sample collection
<ol>
	<li>Measure both food intake and body weight once per week. Fast the mice overnight for a 5-week period.</li>
	<li>Sacrifice them by cervical dislocation and necropsy.</li>
	<li>After the mice have been sacrificed, immobilize them in a supine position.</li>
	<li>Using clean surgical forceps and scissors, locate and lift the central skin near the genitals and make a small 0.2 mm incision.</li>
	<li>Insert the flat scissors horizontally into the incision and carefully separate the abdominal skin from the abdominal wall.</li>
	<li>With the forceps, lift the central skin and with the surgical scissors cut it at the level of the rib cage. Then cut the skin above both of the hind limbs to facilitate the collection of adipose tissue.</li>
	<li>Ensure the collection of all adipose tissue from the body by blunt dissection with scissors and forceps. Once collected, place the adipose tissue in aluminum foil. Remember to collect the fat around the reproductive organs. Avoid the accumulation of hair and skin to not alter the results.</li>
	<li>To access the visceral adipose tissue, cut the abdominal muscle with surgical scissors from the genitals to the rib cage, making an incision in the abdominal muscles from the genitals towards the mouse&#39;s back.</li>
	<li>Perform a hepatectomy to remove 100% of the liver. To externalize the liver, remove unnecessary organs and cut the hepatic veins and arteries to separate the liver from the rest of the body. As the last step, precisely extract the gallbladder from the liver16.</li>
	<li>Measure hypercholesterolemia (cholesterol) and hypertriglyceridemia (triglycerides) using commercial assay kits according to the manufacturer&#39;s indications.</li>
</ol>
5. Statistical analysis
NOTE: All experimental results should be representative from three independent assays, expressed as mean &#177; standard deviation.
<ol>
	<li>Calculate standard error with a one-way ANOVA analysis of variance followed by Tukey&#39;s range test. Consider a P&#60; 0.05 as statistically significant.</li>
</ol>

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Authors declare no conflict of interest.

In this study we evaluated, for the first time, the effect of oral administration of a combination of cloves and cumin (CC) extract on lipid profiles and obesity in mice fed with a high-fat diet for 5 weeks. Findings indicated that the HFD groups showed a significantly higher body weight gain compared to the ND group, which showed that the induction of obesity in the obese model was successful. The administered dose of CC (100, 200 and 450 mg/kg/day) produced a reduction in food intake and body weight. Consequently, the ...

Excess body fat accumulation is a characteristic of obesity. The disequilibrium between energy intake and consumption leads to the storage of excess energy in adipocytes, which is related to metabolic risk factors for hyperglycemia and insulin resistance in type 2 diabetes, hypertension, hypercholesterolemia, and cardiovascular disease1,2.
Natural products with minimum side effects and low cost have received increased attention since previous studies have reported bioactive phytochemicals and potential anti-obesity agents with mechanisms that reverse or delay metabolic syndrome and associated pathologies8.
Several medicinal plants have been studied to prevent obesity and related diseases. Among them, Syzygium aromaticum has been investigated for its anti-obesity potential in in vitro treatment on 3T3-L1 cells and in vivo treatment on mice fed with a high-fat diet11.&#160;In addition, significant anti-overweight effects were observed in a multi-center open trial of Cuminum cyminum on extremely obese subjects12. In this study, C57BL6/J mice was used to investigate the experimental fast model to evaluate a potential anti-obesity agent using a combination of both edible plants.

<table><tbody><tr><td>Distilled Water</td><td>Any vendor</td><td>n/a</td><td>Available for other vendors as well</td></tr><tr><td>Ethanol</td><td>Fermont</td><td>6063</td><td>99.8% purity</td></tr><tr><td>&lt;strong&gt;&amp;nbsp;Diet Ingredients&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Casein</td><td>Any vendor</td><td>n/a</td><td></td></tr><tr><td>Cellulose</td><td>Any vendor</td><td>n/a</td><td></td></tr><tr><td>Centrum balance multivitamin</td><td>Pfizer</td><td>n/a</td><td></td></tr><tr><td>Choline Bitartrate</td><td>Any vendor</td><td>n/a</td><td></td></tr><tr><td>L- cystein</td><td>Sigma Aldrich</td><td>168149</td><td>Available for other vendors as well</td></tr><tr><td>Lard</td><td>Any vendor</td><td>n/a</td><td></td></tr><tr><td>Maltodextrin 10</td><td>Any vendor</td><td>n/a</td><td></td></tr><tr><td>Pellets Nutricubos</td><td>Purina</td><td>n/a</td><td>Available for other vendors as well</td></tr><tr><td>Soybean Oil</td><td>Any vendor</td><td>n/a</td><td></td></tr><tr><td>Sucrose</td><td>Any vendor</td><td>n/a</td><td></td></tr><tr><td>&lt;strong&gt;Extraction Equipment&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Rotavapor</td><td>Buchi</td><td>R-300</td><td></td></tr><tr><td>Shimadzu UV-1800 UV/Visible Scanning Spectrophotometer</td><td>Cole Parmer</td><td>T-83400-20</td><td>Available for other vendors as well</td></tr><tr><td>Ultrasonic Unit</td><td>Elma</td><td>TI-H-20</td><td>Available for other vendors as well</td></tr><tr><td>Vacuum pump</td><td>Buchi</td><td>V-100</td><td></td></tr></tbody></table>

rapid model to evaluate the anti-obesity potential of a combination of syzygium aromaticum (clove) and cuminun cyminum (cumin) on c57bl6/j mice fed high-fat diet

Several studies have demonstrated that the phytochemical contents of plants are potential anti-obesity agents. In this study we examine the effect of using a combination of dry buttons from Syzygium aromaticum and seeds from Cuminum cyminum (CC) on C57BL6/J mice induced with obesity via high-fat-diet (HFD). The aim of this study is to demonstrate that the method proposed in the study reduced obesity significantly after several weeks of experimentation. The extract from both plants was extracted using ultrasound to enhance the extraction of phytochemicals. Optimum extraction conditions were obtained with ethanol as follows: 50:50 v/v water with an ultrasound power of 300 W, and ultrasonication time of 30 minutes. The simultaneous administration of the CC extract in HFD for 5 weeks led to the regulation of lipid profiles (cholesterol and triglycerides), reduction of food intake, weight gain, adipose tissue and liver weight. Findings obtained by this obese model indicate that CC extract can prevent obesity. Compared with the traditional 16 weeks method (8 weeks to get fat, and 8 weeks to lose weight), similar results were obtained in the present study obese model in less time of experimentation.

Ultrasonic-assisted extraction 
The optimum extraction condition for the extraction was ethanol:water (50:50, v/v) with an ultrasound power of 300 W and an ultrasonication time of 30 min. This way, the extraction was faster than conventional extraction methods (Table 2). The extract showed an intense absorption peak at 320 nm in UV-visible spectra (Figure 1).
Food intake and body weight 
The changes in the body weight o...

Watch this Scientific Journal Video about Rapid Model to Evaluate the Anti-Obesity Potential of a Combination of Syzygium aromaticum Clove and Cuminun cyminum Cumin on C57BL6/j Mice Fed High-Fat Diet 

Rapid Model to Evaluate the Anti-Obesity Potential of a Combination of Syzygium aromaticum Clove and Cuminun cyminum Cumin on C57BL6/j Mice Fed High-Fat Diet

This work presents a protocol to explore the anti-obesity effect of two plants used together, for a 5-week duration. There was a combined administration of the extract and high-fat-diet (HFD) in the obese mice. The method can promote the benefits of plants in the treatment of obesity.

Rapid Model to Evaluate the Anti-Obesity Potential of a Combination of Syzygium aromaticum (Clove) and Cuminun cyminum (Cumin) on C57BL6/j Mice Fed High-Fat Diet

Several studies have demonstrated that the phytochemical contents of plants are potential anti-obesity agents. In this study we examine the effect of ...

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Medicine

3.6K Views.  Instituto Politécnico Nacional (IPN) Unidad Profesional Adolfo López Mateos S/N Av. Instituto Politécnico Nacional Ciudad de Mexico. This work presents a protocol to explore the anti-obesity effect of two plants used together, for a 5-week duration. There was a combined administration of the extract and high-fat-diet (HFD) in the obese mice. The method can promote the benefits of plants in the treatment of obesity.

Video: Rapid Model to Evaluate the Anti-Obesity Potential of a Combination of Syzygium aromaticum Clove and Cuminun cyminum Cumin on C57BL6/j Mice Fed High-Fat Diet

Segmentation and Measurement of Fat Volumes in Murine Obesity Models Using X-ray Computed Tomography

Obesity is associated with increased morbidity and mortality as well as reduced metrics in quality of life.1 Both environmental and genetic factors are associated with obesity, though the precise underlying mechanisms that contribute to the disease are currently being delineated.2,3 Several small animal models of obesity have been developed and are employed in a variety of studies.4 A critical component to these experiments involves the collection of regional and/or total animal fat content data under varied conditions.
Traditional experimental methods available for measuring fat content in small animal models of obesity include invasive (e.g. ex vivo measurement of fat deposits) and non-invasive (e.g. Dual Energy X-ray Absorptiometry (DEXA), or Magnetic Resonance (MR)) protocols, each of which presents relative trade-offs. Current invasive methods for measuring fat content may provide details for organ and region specific fat distribution, but sacrificing the subjects will preclude longitudinal assessments. Conversely, current non-invasive strategies provide limited details for organ and region specific fat distribution, but enable valuable longitudinal assessment. With the advent of dedicated small animal X-ray computed tomography (CT) systems and customized analytical procedures, both organ and region specific analysis of fat distribution and longitudinal profiling may be possible. Recent reports have validated the use of CT for in vivo longitudinal imaging of adiposity in living mice.5,6 Here we provide a modified method that allows for fat/total volume measurement, analysis and visualization utilizing the Carestream Molecular Imaging Albira CT system in conjunction with PMOD and Volview software packages.

Fat content analysis is routinely conducted in studies utilizing murine obesity models. Emerging methods in small animal CT imaging and analysis are providing for longitudinal detail rich fat content analysis. Here we detail step by step procedures for performing small animal CT imaging, analysis, and visualization.

Obesity is associated with increased morbidity and mortality as well as reduced metrics in quality of life.1 Both environmental and genetic factors are ...

Evaluating the Anti-depression Effect of Xiaoyaosan on Chronically-stressed Mice

In addition to the standardized use of antidepressant medications and psychotherapy, the usage of traditional Chinese medicine has lead to an overall improvement of patients with major depressive disorder (MDD). Therefore, the purpose of this study was to establish the mouse depressive model, observe the behavior changes associated with chronic unpredictable mild stress (CUMS), and then evaluate the anti-depression effect of Xiaoyaosan. Mice were randomly divided into four groups: a control group, a model group, a treatment group with Xiaoyaosan, and a treatment group with fluoxetine. All mice were individually kept in cages, and depression was induced in the mice by exposing them to several designed manipulations of CUMS for 21 days, as described in the protocol. Mice in the control group and model group received 0.5 mL of distilled water, while mice in the treatment groups received either Xiaoyaosan (0.25 g/kg/day) or fluoxetine (2.6 mg/kg/day). The drugs used in the study were given intragastrically daily during the entire three weeks. To estimate the depressive-like behaviors, a series of parameters including the coat state, body weight, open field test score, and sucrose preference test score were recorded. Data analysis showed that behaviors of model mice were significantly changed compared to behaviors of mice in the control group, which were improved by the treatment of Xiaoyaosan and fluoxetine. The current findings demonstrated the anti-depression effects of Xiaoyaosan on the behaviors of CUMS-induced mice and revealed that compounds from the Xiaoyaosan prescription may be worthwhile for treating depression, considering their beneficial effects on depressive-like behaviors.

Here, we present a protocol to establish the mouse depressive model, observe the behavior changes associated with chronic unpredictable mild stress (CUMS), and evaluate the anti-depression effect of Xiaoyaosan.

In addition to the standardized use of antidepressant medications and psychotherapy, the usage of traditional Chinese medicine has lead to an overall ...

Behavior

Optimized Analysis of In Vivo and In Vitro Hepatic Steatosis

Establishing a system of procedures to qualitatively and quantitatively characterize in vivo and in vitro hepatic steatosis is important for metabolic study in the liver. Here, numerous assays are described to comprehensively measure the phenotype and parameters of hepatic steatosis in mouse and hepatocyte models. 
Combining the physiological, histological, and biochemical methods, this system can be used to assess the progress of hepatic steatosis. In vivo, the measurements of body weight and nuclear magnetic resonance (NMR) provide a general understanding of mice in a non-invasive manner. Hematoxylin and Eosin (H&amp;E) and Oil Red O staining determine the histological morphology and lipid deposition of liver tissue under nutrient overload conditions, such as high-fat diet feeding. Next, the total lipid contents are isolated by chloroform/methanol extraction, which are followed by a biochemical analysis for triglyceride and cholesterol. Moreover, mouse primary hepatocytes are treated with high glucose plus insulin to stimulate lipid accumulation, an efficient in vitro model to mimic diet-induced hyperglycemia and hyperinsulinemia in vivo. Then, the lipid deposition is measured by Oil Red O staining and chloroform/methanol extraction. Oil Red O staining determines intuitive hepatic steatotic phenotypes, while the lipid extraction analysis determines the parameters that can be analyzed statistically. The present protocols are of interest to scientists in the fields of fatty liver diseases, insulin resistance, and type 2 diabetes.

Optimized Analysis of In Vivo and In Vitro Hepatic Steatosis

Here, optimized methods to generate in vivo and in vitro models of hepatic steatosis and to analyze the steatotic phenotypes and related physiological parameters are described.

Establishing a system of procedures to qualitatively and quantitatively characterize in vivo and in vitro hepatic steatosis is important for metabolic ...

Study of In Vivo Glucose Metabolism in High-fat Diet-fed Mice Using Oral Glucose Tolerance Test (OGTT) and Insulin Tolerance Test (ITT)

Obesity represents the most important single risk factor in the pathogenesis of type 2 diabetes, a disease which is characterized by a resistance to insulin-stimulated glucose uptake and a gross decompensation of systemic glucose metabolism. Despite considerable progress in the understanding of glucose metabolism, the molecular mechanisms of its regulation in health and disease remain under-investigated, while novel approaches to prevent and treat diabetes are urgently needed. Diet derived glucose stimulates the pancreatic secretion of insulin, which serves as the principal regulator of cellular anabolic processes during the fed-state and thus balances blood glucose levels to maintain systemic energy status. Chronic overfeeding triggers meta-inflammation, which leads to alterations in peripheral insulin receptor-associated signaling and thus reduces the sensitivity to insulin-mediated glucose disposal. These events ultimately result in elevated fasting glucose and insulin levels as well as a reduction in glucose tolerance, which in turn serve as important indicators of insulin resistance. Here, we present a protocol for the generation and metabolic characterization of high-fat diet (HFD)-fed mice as a frequently used model of diet-induced insulin resistance. We illustrate in detail the oral glucose tolerance test (OGTT), which monitors the peripheral disposal of an orally administered glucose load and insulin secretion over time. Additionally, we present a protocol for the insulin tolerance test (ITT) to monitor whole-body insulin action. Together, these methods and their downstream applications represent powerful tools to characterize the general metabolic phenotype of mice as well as to specifically assess alterations in glucose metabolism. They may be especially useful in the broad research field of insulin resistance, diabetes and obesity to provide a better understanding of pathogenesis as well as to test the effects of therapeutic interventions.

Study of In Vivo Glucose Metabolism in High-fat Diet-fed Mice Using Oral Glucose Tolerance Test OGTT and Insulin Tolerance Test ITT

The current article describes the generation and metabolic characterization of high-fat diet-fed mice as a model of diet-induced insulin resistance and obesity. It further features detailed protocols to perform the oral glucose tolerance test and the insulin tolerance test, monitoring whole-body alterations of glucose metabolism in vivo.

Obesity represents the most important single risk factor in the pathogenesis of type 2 diabetes, a disease which is characterized by a resistance to ...

Body Composition and Metabolic Caging Analysis in High Fat Fed Mice

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate utilization, and behaviors such as food intake and physical activity can provide important information regarding the underlying mechanisms of disease. Given the importance of body composition and metabolism to the development of obesity and its subsequent sequelae, it is necessary to make accurate measures of these parameters in the pre-clinical research setting. Advances in technology over the past few decades have made it possible to derive these measures in rodent models in a non-invasive and longitudinal fashion. Consequently, these metabolic measures have proven useful when assessing the response of genetic manipulations (for example knockout or transgenic mice, viral knock-down or overexpression of genes), experimental drug/compound screening and dietary, behavioral or physical activity interventions. Herein, we describe the protocols used to measure body composition and metabolic parameters using an animal monitoring system in chow-fed and high fat diet-fed mice.

This protocol describes the use of a body composition analyzer and metabolic animal monitoring system to characterize body composition and metabolic parameters in mice. An obesity model induced by high-fat feeding is used as an example for the application of these techniques.

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate ...

Palatable Western-style Cafeteria Diet as a Reliable Method for Modeling Diet-induced Obesity in Rodents

Obesity is rapidly increasing in incidence in developed and developing countries and is known to induce or exacerbate many diseases.&#160;The health burden of obesity and its comorbid conditions highlight the need for better understanding of its pathogenesis, yet ethical constraints limit studies in humans. To this end externally valid models of obesity in laboratory animals are essential for the understanding of being overweight and obesity.&#160;While many species have been used to model the range of changes that accompany obesity in humans, rodents are most commonly used. Our laboratory has developed a western-style cafeteria diet that consistently leads to considerable weight gain and markers of metabolic disease in rodents. The diet exposes rodents to a variety of highly palatable foods to induce hyperphagia, modeling the modern western food environment. This diet rapidly induces weight gain and body fat accumulation in rats allowing for the study of effects of overeating and obesity. While the cafeteria diet may not provide the same control over macronutrient and micronutrient profile as purified high-fat or high-fat, high-sugar diets, the cafeteria diet typically induces a more severe metabolic phenotype than that observed with purified diets and is more in line with metabolic disturbances observed in the overweight and obese human population.

This protocol describes the use of a highly palatable, western-style cafeteria diet to model overeating and obesity in rodents. Here, we provide a detailed outline of food selection, preparation and measurement, and explain methodological factors that assist in generating a robust and reproducible phenotype.

Obesity is rapidly increasing in incidence in developed and developing countries and is known to induce or exacerbate many diseases.&#160;The health ...

Assessing Whole-Body Lipid-Handling Capacity in Mice

Assessing lipid metabolism is a cornerstone of evaluating metabolic function, and it is considered essential for in vivo metabolism studies. Lipids are a class of many different molecules with many pathways involved in their synthesis and metabolism. A starting point for evaluating lipid hemostasis for nutrition and obesity research is needed. This paper describes three easy and accessible methods that require little expertise or practice to master, and that can be adapted by most labs to screen for lipid-metabolism abnormalities in mice. These methods are (1) measuring several fasting serum lipid molecules using commercial kits (2) assaying for dietary lipid-handling capability through an oral intralipid tolerance test, and (3) evaluating the response to a pharmaceutical compound, CL 316,243, in mice. Together, these methods will provide a high-level overview of lipid handling capability in mice.

This paper provides three easy and accessible assays for assessing lipid metabolism in mice.

Assessing lipid metabolism is a cornerstone of evaluating metabolic function, and it is considered essential for in vivo metabolism studies. Lipids are a ...

Network Pharmacology Prediction and Metabolomics Validation of the Mechanism of Fructus Phyllanthi against Hyperlipidemia

Hyperlipidemia has become a leading risk factor for cardiovascular diseases and liver injury worldwide. Fructus Phyllanthi (FP) is an effective drug against hyperlipidemia in Traditional Chinese Medicine (TCM) and Indian Medicine theories, however the potential mechanism requires further exploration. The present research aims to reveal the mechanism of FP against hyperlipidemia based on an integrated strategy combining network pharmacology prediction with metabolomics validation. A high-fat diet (HFD)-induced mice model was established by evaluating the plasma lipid levels, including total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C). Network pharmacology was applied to find out the active ingredients of FP and potential targets against hyperlipidemia. Metabolomics of plasma and liver were performed to identify differential metabolites and their corresponding pathways among the normal group, model group, and intervention group. The relationship between network pharmacology and metabolomics was further constructed to obtain a comprehensive view of the process of FP against hyperlipidemia. The obtained key target proteins were verified by molecular docking. These results reflected that FP improved the plasma lipid levels and liver injury of hyperlipidemia induced by a HFD. Gallic acid, quercetin, and beta-sitosterol in FP were demonstrated as the key active compounds. A total of 16 and six potential differential metabolites in plasma and liver, respectively, were found to be involved in the therapeutic effects of FP against hyperlipidemia by metabolomics. Further, integration analysis indicated that the intervention effects were associated with CYP1A1, AChE, and MGAM, as well as the adjustment of L-kynurenine, corticosterone, acetylcholine, and raffinose, mainly involving tryptophan metabolism pathway. Molecular docking ensured that the above ingredients acting on hyperlipidemia-related protein targets played a key role in lowering lipids. In summary, this research provided a new possibility for preventing and treating hyperlipidemia.

The present protocol describes an integrated strategy for exploring the key targets and mechanisms of Fructus Phyllanthi against hyperlipidemia based on network pharmacology prediction and metabolomics verification.

Hyperlipidemia has become a leading risk factor for cardiovascular diseases and liver injury worldwide. Fructus Phyllanthi (FP) is an effective drug ...

Assessment of the Metabolic Effects of Isocaloric 2:1 Intermittent Fasting in Mice

Intermittent fasting (IF), a dietary intervention involving periodic energy restriction, has been considered to provide numerous benefits and counteract metabolic abnormalities. So far, different types of IF models with varying durations of fasting and feeding periods have been documented. However, interpreting the outcomes is challenging, as many of these models involve multifactorial contributions from both time- and calorie-restriction strategies. For example, the alternate day fasting model, often used as a rodent IF regimen, can result in underfeeding, suggesting that health benefits from this intervention are likely mediated via both caloric restriction and fasting-refeeding cycles. Recently, it has been successfully demonstrated that 2:1 IF, comprising 1 day of fasting followed by 2 days of feeding, can provide protection against diet-induced obesity and metabolic improvements without a reduction in overall caloric intake. Presented here is a protocol of this isocaloric 2:1 IF intervention in mice. Also described is a pair-feeding (PF) protocol required to examine a mouse model with altered eating behaviors, such as hyperphagia. Using the 2:1 IF regimen, it is demonstrated that isocaloric IF leads to reduced body weight gain, improved glucose homeostasis, and elevated energy expenditure. Thus, this regimen may be useful to investigate the health impacts of IF on various disease conditions.

The current article describes a detailed protocol for isocaloric 2:1 intermittent fasting to protect and treat against obesity and impaired glucose metabolism in wild-type and ob/ob mice.

Intermittent fasting (IF), a dietary intervention involving periodic energy restriction, has been considered to provide numerous benefits and counteract ...

Biology

Murine Model for Diabetic Cardiomyopathy via High-Fat Diet and STZ Injection

Modeling and Evaluation of Murine Diabetic Cardiomyopathy Model

The underlying pathophysiological mechanisms of diabetic cardiomyopathy (DbCM), a leading cause of mortality among patients with type 2 diabetes mellitus (T2DM), remain poorly understood. The myocardial toxicity associated with T2DM is attributed to factors such as lipotoxicity, glucotoxicity, oxidative stress, reduced cardiac efficiency, and lipoapoptosis. Compared to rats, mice offer greater accessibility, cost-effectiveness, and broader applicability for animal experiments. Insulin resistance and impaired insulin secretion are crucial factors in the pathophysiology of T2DM. We introduce a novel nongenetic murine model that replicates the progression of human DbCM induced by a combination of high-fat diet (HFD) feeding and streptozotocin (STZ) injection. In this study, we used wild-type C57BL/6J mice, administering an HFD regimen for 12 weeks, followed by intraperitoneal injections of STZ for an additional 12 weeks to induce characteristic manifestations of T2DM. We conducted oral glucose tolerance tests and measured serum insulin concentrations to confirm the development of insulin resistance and insufficient insulin secretion. Cardiac structure and function were rigorously assessed through noninvasive transthoracic echocardiography. Pathological characteristics were evaluated through Masson's trichrome staining and wheat germ agglutinin (WGA) staining, revealing pathological features related to DbCM. Therefore, we provide a robust and versatile method for establishing a nongenetic murine model of DbCM.

This protocol outlines a method for inducing diabetic cardiomyopathy through a combination of high-fat diet feeding and streptozotocin injection. This approach aims to provide a reliable framework for scientific investigation into diabetic cardiomyopathy and to explore potential avenues for clinical treatment applications.

The underlying pathophysiological mechanisms of diabetic cardiomyopathy (DbCM), a leading cause of mortality among patients with type 2 diabetes mellitus ...

Rapid Model to Evaluate the Anti-Obesity Potential of a Combination of Syzygium aromaticum (Clove) and Cuminun cyminum (Cumin) on C57BL6/j Mice Fed High-Fat Diet

Summary

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Optimized Analysis of In Vivo and In Vitro Hepatic Steatosis

Study of In Vivo Glucose Metabolism in High-fat Diet-fed Mice Using Oral Glucose Tolerance Test (OGTT) and Insulin Tolerance Test (ITT)

Body Composition and Metabolic Caging Analysis in High Fat Fed Mice

Palatable Western-style Cafeteria Diet as a Reliable Method for Modeling Diet-induced Obesity in Rodents

Assessing Whole-Body Lipid-Handling Capacity in Mice

Network Pharmacology Prediction and Metabolomics Validation of the Mechanism of Fructus Phyllanthi against Hyperlipidemia

Assessment of the Metabolic Effects of Isocaloric 2:1 Intermittent Fasting in Mice

Modeling and Evaluation of Murine Diabetic Cardiomyopathy Model