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

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.

Abstract

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.

Introduction

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 disease^1,2.

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 ....

Protocol

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.

1. 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 ± 4 °C.¹³

Results

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 of th.......

Discussion

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 .......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Distilled Water	Any vendor	n/a	Available for other vendors as well
Ethanol	Fermont	6063	99.8% purity
Diet Ingredients
Casein	Any vendor	n/a
Cellulose	Any vendor	n/a
Centrum balance multivitamin	Pfizer	n/a
Choline Bitartrate	Any vendor	n/a
L- cystein	Sigma Aldrich	168149	Available for other vendors as well
Lard	Any vendor	n/a
Maltodextrin 10	Any vendor	n/a
Pellets Nutricubos	Purina	n/a	Available for other vendors as well
Soybean Oil	Any vendor	n/a
Sucrose	Any vendor	n/a
Extraction Equipment
Rotavapor	Buchi	R-300
Shimadzu UV-1800 UV/Visible Scanning Spectrophotometer	Cole Parmer	T-83400-20	Available for other vendors as well
Ultrasonic Unit	Elma	TI-H-20	Available for other vendors as well
Vacuum pump	Buchi	V-100