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 five-week duration. There was a combined administration of the extract and high-fat diet in the obese mice. The method can promote the benefits of plants in the treatment of obesity.

Excessive modified accumulation is characteristic of obesity. between energy intake and expenditure. Let's do the storage of excess of energy in adipocytes.

Actually, obesity is the cause of the 78%of type two diabetes, the 68%of hypertensive disease, and the 45%of ischemic stroke cases. Globally, it has been predicted that there will be 2, 000, 000 overweight and 1, 000, 000 obese individuals by 2030. Evidence for anti-obesity drugs indicated a high incidence of adverse effects and toxicity.

Surgical treatment intervention, such as bariatric surgery, are not always treatable. Consequently, more studies are needed to develop new treatments that could be used to decrease the prevalence of obesity. Medicinal plants contain pharmacodynamic bioactive compounds whose synergistic effect is beneficial in the management of metabolic disorders.

The discovery of these phytochemical components, and their use reduce the financial burden of relying on inexpensive synthetic drugs. Another driving factor in the use of medicinal plants is the perception that they are devoid of adverse side effects. Therefore, the study of medicinal plants and their biotic compounds has emerged as a key development in modern medicine.

Several medicinal plants have been studied to avoid obesity and related diseases. Among them, Syzygium aromaticum has been investigated for its anti-obesity potential in mutual treatments. Significant anti-overweight effects were observed in a multicenter open trial of Cuminum cyminum in extremely obese subjects.

And this study, we use C, 57, B, L, six, J mice to investigate an experimental first model to evaluate a potential anti-obesity agent for reduced obesity. The edible plants Syzygium aromaticum, clove, and Cuminum cyminum, cumin, were purchased from Mexico. Performing the extraction procedures through ultrasonic assisted extraction with the grinding of the seeds of the two plants.

500 grams. Sonic-ate them with ethanol. Water, 50 to 50, V over V, at a temperature range of 30 plus minus four degrees Celsius.

Later, filter the solutes using Whatman number two filter paper under vacuum, and concentrate the extract with a rotary evaporator. Then the UV vis spectrum of the extract of both plants was measured. Male C, 57, B, L, six, J mice of six weeks old were used for the study.

The animals were housed in groups of eight in standard laboratory conditions of temperature, relative humidity, lighting, with food and water ad linium. The mice were acclimatized in these conditions for a week before the experiment. All animal experiments were approved by the Animal Experiment Comity of A kilogram of the high fat diet used in this experiment is prepared with 140 grams of casein, one point eight grams of L cysteine, 120 grams of lard, 40 grams of soybean oil, 150 grams of Maltodextrin 10, 450 grams of sucrose, 50 grams of cellulose, a tablet equivalent of vitamin L and minerals, and two point five grams of Choline bitartrate.

At the beginning of the test, subject each one of the groups form to different treatments. Feed group one on a normal diet, group two on a high-fat diet, group three on a high fat diet plus 100 milligrams over kilograms of C, C extract, group four on a high-fat diet plus 250 milligrams over kilograms of C, C extract, group five on a high-fat diet plus 450 milligrams over kilograms of C, C extract, group six on a high-fat diet plus phentermine, an anti-obesity drug used as positive control. Feed all the groups of mice for five weeks and record the amount of food consumed daily.

For this, in a test tube with stopper, the extract or the drug is homogenized with a dose of water corresponding to the group to which the treatment is being administered. This is done with vortex shaker. Once homogenized, the corresponding treatment is emptied into the drinker of each group.

Measure both food intake and body weight once per week. Fast the mice overnight for a five week period, then sacrifice them by cervical dislocation and necropsy. After the mice have been sacrificed, immobilize them in a supine position.

Using clean surgical forceps and scissors, locate and lift the central skin near the genitals and make a small point two millimeter incision. Insert the flat scissors horizontally into the incision and carefully separate the abdominal skin from the abdominal wall. 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 the adipose tissue. Ensure collection of all adipose tissue from the body by blond to section with scissors and forceps, and 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, so as not to alter the results. To access visceral adipose tissue, cut to the abdominal muscle with surgical scissors from the genitals through the rib cage, making an incision in the abdominal muscles from the genitals towards the mouse's back. A hepatectomy is performed to remove a hundred percent of the liver.

To externalize the liver, unnecessary organs are removed and the hepatic veins and arteries are cut to separate the liver from the rest of the body. As the last step, extract precisely the gallbladder from the liver. Measure hypercholesterolemia and hypertriglyceridemia using commercial ELISA kits, according to the manufacturer's indications.

All experimental results should be representative from three independent essays. Expressed us mean plus minus standard deviation. Calculate standard error with a one way ANOVA analysis of variance followed by Tukey honest.

Let significant difference of P less than zero point zero five be considered as statistically significant. Optimum extraction conditions were obtained with ethanol as follows 50 to 50 V over V water with an ultrasound power of 300 W.Ultrasonic-cation time of 30 minutes. This way the extraction was faster than the conventional extraction methods, table two.

The extract showed an intense absorption peak at 320 N, M, in UV visible spectrum, figure one. Food intake and body weight. The changes in the body weight of the mice given different concentrations of the extract.

100, 200 and 450 milligrams over kilograms for five weeks are shown in figure two, A.The percentages of weight reduction in groups three, four, and five were 12.2, 30, and 41 percent respectively. This shows the lowest body weight gain was observed in older groups, but the HFD group had an increase of two point one fold compared to the normal fat diet group. This data indicates that 450 milligrams over kilograms dose of C, C is more effective than 30 milligrams over kilograms phentermine.

39%reduction in reducing body weight gain. The main food intake at the end of the experiment in the HFD plus C, C was 31%lower than the in the obese group. With one point three grams reduction in food intake compared to the obese group.

Food intake in the HFD fed mice was higher by one point five fold than in the normal control. Thus, it's significantly decreased in all C, C groups in a range of 7.14 to 31%The diet efficiency rate in HFD group was 16%which was significantly different from that of the control group at 11%These results demonstrate that C, C can reduce HFD induced body weight gain. The weight of Epididymal WAT, subcutaneous fat, and Perirenal WAT adipose tissues was higher in the HFD group, but not in the HFD, C, C groups.

The liver weight was significantly decreased in the C, C group, with a range of 15 to 28.4 percent, compared to those of the HFD group. The levels of serum triacylglycerol, TG, and total cholesterol, TC, after five weeks of treatment are shown in figures three, A and three, B, respectively. The triglycerides and total cholesterol level were significantly elevated by one point three two, and one point three one, respectively, in the HFD groups compared to those in the ND groups.

The increased levels of plasma TG and TC in the HFD diet were significantly attenuated by the administration of 200 and 450 milligrams per kilograms per day, C, C extract by 18.18 and 22.7%for TG and 16.41 and 20.61%for TC respectively compared to the control. In this study, we made the first model to determine the anti-obesity effect of an extract from a combination of two edible plants, Syzygium aromaticum, clove, and Cuminum cyminum, cumin, in C, 57, B, L, six, J mice, which roughly developed obesity and ameliorated hyperlipidemia induced by a high-fat diet feeding. After verifying findings in this research provides solid evidence that these first five weeks of duration could be used to assess whether a plant has capacity as a potential agent for reduce obesity.