The Effect of Capsaicin on the Metabolic Rate of Male Mus Musculus

The Effect of Capsaicin on the Metabolic Rate of Male Mus musculus

Linda Ko

Department of Biological Sciences

Saddleback College Mission Viejo, CA, 92692

(Title centered)

Capsaicin (8-methyl-N-vanillyl-6-nonenamide) is the chemical component in chili peppers that gives them heat(that produces a spicy sensation). The purpose of this experiment was to examine the effect of capsaicin on the metabolic rate in male Mus musculus. To calculate the metabolic rate, the production of CO2CO2iswasused. The objective of the study was to see what the impact of capsaicin would have on the basal metabolic rates (BMR) of mice. Twelve mice were used at different times to examine the BMR of mice with no substances, a control and experimental solution(How many mice at each time? Or did each time have 12 studies with a total of 36 mice? Or did you use all 12 for each trial??)The control and experimental groups were stomach fed via a gavage tube. They were then put into a CO2 CO2 analyzer. The hypothesis being tested is wasthat the consumption of capsaicin would have an effectaffect on the metabolic rates of miceMus musculus. The average BMR of the mice without any substances was 0.419 mLCO2 CO2/g/min (±0.0310 SEM, N=12). The average BMR of mice with the control solution was 0.230mLCO2 CO2 /g/min (±0.0284 SEM, N=12). The average BMR of mice with the experimental capsaicin solution was 0.341mLCO2CO2 /g/min (±0.0149 SEM, N=12). A One-Way Analysis of Variance test was used and the results indicate that there was a significant difference in the metabolic rate between the subject groups (p<0.0001). Tukey’s HSD Post Hoc test showed significant differences between all groups (p<0.05).

Capsaicin (8-methyl-N-vanillyl-6-nonenamide) is the chemical component in chili peppers that produces a spicy sensation. The objective of the study was to test the impact of capsaicin on the basal metabolic rates (BMR) of mice, Mus musculus. To calculate the metabolic rate, the production of CO2 was used. A total of twelve mice were separated into 3 groups of 4 mice at different times to examine the BMR of mice with no substances, control group and experimental solution???. The control and experimental groups were stomach fed via a gavage tube. They were then put into a CO2 analyzer. The hypothesis being tested was that the consumption of capsaicin would have an affect on the metabolic rates of Mus musculus. The average BMR of the mice without any substances was 0.419 mL CO2/g/min (±0.0310 SEM, N=12). The average BMR of mice with the control solution was 0.230mL CO2 /g/min (±0.0284 SEM, N=12). The average BMR of mice with the experimental capsaicin solution was 0.341mL CO2 /g/min (±0.0149 SEM, N=12). A One-Way Analysis of Variance test was used which resulted in a significan difference in metabolic rate between the subject groups (p<0.0001). Tukey’s HSD Post Hoc test showed significant differences between all groups (p<0.05, one-tailed or two-tailed????).

Introduction

Capsaicin (C18H27NO3)(C18H27NO3), a chemical compound (8- methyl-N-vanillyl-6-nonenamide), is the key component in red chili peppers that gives your the(try to not use first or second person)mouth a kick once it’s it is consumed (Hursel and Westerterp-Plantenga, 2009). This chemical compound is extremely pungent and irritates the skins of humans and animals. Capsaicin is notably found as a major pungent factor in the (take out the)fruits of Capsicum and it (take out it)has been stated that capsaicin increases the catecholamine secretion, energy outflow, and also reduces body fat in perspective of a long term treatment (Ohnuki et al., 2001). Capsaicin, a major component in red hot peppers, is a trigger for the bodytriggers the body to undergo thermogenesis. Thermogenesis is a process in which the body temperature is increased due to many biochemical and metabolic events. (Kobayashi et al., 1998) With an increase in thermogenesis, the body’s metabolic rates are raisedmetabolic rate increases; therefore, stored fat cells in the body are utilized as energy to support the body’s increased metabolic rate, resulting in weight loss.

In a previously published (do not really need published, so it would be In a previous study) study done by Ohnuki and his team (2001) at Kyoto University, researchers were able to see the result of M. musculus loss of weight were able to observe the weight loss of M. musculus.Their study was successful; however, we differ in the intentions of our study (???????????????????????). They were examining the long term effects of capsaicin on weight loss, and wewere studying the effects on their metabolic rates(NO! THIRD PERSON!!!). Ohnuki and his team colleagues (2001) also gave the mice CH-19 Sweet, the fruit of a non-pungent pepper, capsaicin that contains small amounts of capsaicinoids (Hursel and Westerterp-Plantenga, 2009; Kawabata et al., 2009; Ohnuki et al.,2001)(Ohnuki et al.,2001; Hursel and Westerterp-Plantenga, 2009; Kawabata et al., 2009). Cited references- if there is more than one, then it needs to be in chronological order.

The objective was to determine if the basal metabolic rates (BMR) of the mice would be affected when given capsaicinof mice effected when given capsaicin.BMR is the metabolic rate of a fasting mammal resting at a normal temperature in a dark room, in a conscious state (Does not make sense. Please reword).The hypothesis being tested is that the mice given the capsaicin would have an effect on BMR compared to the two other groupsThe hypothesis was tested to observe the effect on metabolic rate after the consumption of capsaicin.

Materials andMethods

Twelve male M. musculus were purchased and used as both the control and experimental group. They were kept in the guest bathroom at Linda Ko’sof the investigators house where they remained for 8 days prior to the start of the experiment. This was done to give them ample time to adapt to their environment and stabilize. The cages were placed in an isolated enclosure to prevent them from being scared by outside environmental factors. The room was blocked off from natural light; instead lights were turned on for 12 hours a day. Due to the mice being of a circadian species, the tests were given during times between 11:00 to 16:00 to avoid disturbances. They were given fresh food and water daily. Three hours prior to the runs with the CO2 CO2 analyzer, their food was taken away in to prevent any factors that may have arisen. Each mouse was weighed, and then had various colored rings drawn on their tails for identification. The CO2 CO2 analyzer had air flowing through the first chamber, where the mouse was contained. It then went down through a tube containing Drierite (Xenia, Ohio), then into the last chamber where a Pasco GLX Carbon Dioxide Gas sensor was placed. The sensor was connected to a Pasco GLX unit that generated a graph. A Pasco GLX barometer and temperature sensors were also used. A FoxBox (Sable Systems, Las Vegas) was used to generate flow rate (Figure 1).

The first trials weretrial of 12? or 4? micethe mice without any substances. A time of one minute was used to stabilize the machine. TheOne or Amouse(how many???) was put into the first tube and remained inside the tube for five minutes with a towel covering it. The mouse was then taken out and the investigators had to wait until the graph dropped down to the stabilization rate. It was then recorded for another minute. Calculations for the amount of solution given to the mice were done prior to stomach feeding them. Shown in a previous study, 10 mg of solution was distributed per kg body weight. The control solution consisted of 3 mL ethanol, 10 mL Polysorbate 80 (Tween 80) and enough NaCl to make 100 mL (Ohnuki et al. 2001, Kawabata et al. 2009)No need for this!!! This section is where you talk about what you did. The experimental solution consisted of 1.0 mg capsaicin, 3 mL ethanol, 10 mL Polysorbate 80 (Tween 80) and enough NaCl to make a total volume of 100 mL. The pure capsaicin was purchased from Sigma Aldrich. The mice were fed their control and experimental solution (What was their control solution???)via a gavage tube and then tested one hour after all of them had been fed. Six stainless steel gavage tubes were purchased from Instech Laboratories, model FTSS- 18S-51. The Institutional Animal Care and Use Committee (IACUC) at the University of California, San Francisco, states the standard procedure of oral gavage in mice is done by appropriately inserting the gavage tube into the mice. The length from the tip of the head to the last rib must be determined in order to prevent puncturing a hole in their stomach. A mouse must be scruffed by grasping its skin over the shoulders, utilizing the researcher’s thumb, middle finger, and pinky specifically for holding the tail. The fore legs must be extended out to the side in to prevent them kicking the tube away. The index finger will be placed on top of the head to bring it back. Doing so will create a straight line through the neck and esophagus. (You have to talk about what you did not what other studies have done. You explained this in the next couple sentences, so the highlighted blue is not necessary).The gavagetube was placed on either sideWhat side? of the mouse, but made sure it was over the tongue. The tube went through the pharynx, while pressing the gavage tube to the roof of the mouth, causing the mouth to elevate the head and the esophagus to straighten. Put the tube inThe tube was placed….WHERE?. The tube was taken out the same way put in. The data found was used to calculate the rate of CO2 CO2 produced and amount of O2 O2 consumed.

The rate of carbon dioxide production was calculated using the following equation:

VCO2= (Ve-V1)(Flow Rate mL/min).(1) What does this one represent???

This volume was correct to STPD using Boyle’s law. Assuming a respiratory quotient of 0.8, we converted rate of carbon dioxide consumption t=into oxygen consumption. The rate of carbon dioxide consumption was converted to oxygen consumption.All rates were divided by animal weight to give weight specific data. (Was it divided by the average weight or the specific weight to the rate for that specific rat? If so please reword to make it clear).

Figure 1. System for measuring CO2  CO2 production.

55 Saddleback Journal of Biology Vol. 9, Spring 2011. (Font Size same as the rest of the paper)

Results

The end result turned out that there was a statistically significant difference between the three groups, which consist of the same group of male mice, without any substances, control solution, and experimental solution. The mice without any of the substances showed a mean of 7.74 mLCO2 CO2/min (± 0.0310 SEM, N=12). The mice with the 10% Polysorbate 80 in 0.9% NaCl solution had a mean of 4.32 mLCO2 CO2/min (± 0.0284 SEM, N=12). AndThe mice that waswere given the 0.1mg/mL Capsaicin mixed with 10% Polysorbate 80 in 0.9% NaCl had a mean of 6.50 mLCO2 /min (±0.0149 SEM, N=12). Therefore, the mice that were given the Capsaicin solution resulted in a significant difference (p=0.00372, one-tailed or two-tailed???) with a PostHoc of Mean (M) 1 vs. M2 (p<0.01), M1 vs. M3 (nonsignificant), M2 vs. M3 (p<0.05).

WhenThe BMR was calculated, the mean of the mice without any substances turned out to be 0.419 mLCO2/g/min (± 0.0310 SEM, N=12), the mice given the control solution turned out to have a mean of 0.230 mLCO2/g/min (± 0.0284 SEM, N=12), and the mice given the experimental solution had a mean of 0.341 mLCO2/g/min (± 0.0149 SEM, N=12). There was a statistically significant difference (p<0.0001, ANOVA). PostHoc: M1 vs. M2 (p<0.01), M1 vs. M3 (nonsignificant), M2 vs. M3 (p<0.01).

In order to show the relations of the different groups a line graph and a bar graph was constructed (Figure 2 and Figure 3).  Does not need to be bolded.

(Center the Graph)

Figure 2. BMR measurements of each mouse for every run (p<0.0001, ANOVA. PostHoc: M1 vs. M2 (p<0.01), M1 vs. M3 (nonsignificant), M2 vs. M3 (p<0.01)).(Font Size same as the rest of the paper)

(Center the Graph and Take off the Line of the Graph)

Figure 3. The average BMR of the mice of the three groups (none 0.419 mLCO2/g/min (± 0.0310 SEM, N=12), control solution 0.230 mLCO2/g/min (± 0.0284 SEM, N=12), experimental solution 0.341. add Error bars indicate mean ± SEM. (Font Size same as the rest of the paper)

Discussion

The study demonstrated that the hypothesis tested was proved wrong even though there was a significant difference. Capsaicin does increase the BMR of the mice, however, the group without any substances had an elevated BMR, overriding the capsaicin group. As seen in Figure 2, the group that had no substances had a higher BMR value than the group with capsaicin. The investigators suggest that the reason why this occurred is because the run, where nothing was given, was the mouse’s first time in the constricted chamber and the mice may have panicked. In Ohunki’sexperiment(YEAR), he and his team had an increase in oxygen consumption the oxygen consumption increased for the group that was given capsaicin (p<0.05). In the experiment conducted by the investigators, they too got a significant difference ofO2 O2consumption (p=4.74 x 10-4), and also forCO2 CO2production (p=3.72 x 10-3) and BMR (p<0.0001).(Reword. Jumping from a study years ago to this experiment. It is a little confusing.)P values were analyzed to determine if there is a significant difference. In order to run the values under ANOVA, each mouse did all three runs, but not all in the same day.

In future experiments, the investigators should make sure that the equipment’s being used for their project are working properly and let the mice be accustomed to the chamber beforehand. (Cannot write this because this suggests that you could have done another trial with correcting these errors)

For the discussion, explain why you got the results instead of restating what you got.

Acknowledgements

Iwould like to thank Professor Steve Teh and Dr. Tony Huntley for all of their help during this project.

Literature Cited

Hursel, R, Westerterp-Platenga, MS. Thermogenic ingredients and body weight regulation. International Journal of Obesity. (2010). (34):659-669.

Kawabata F, Inoue N, Masamoto Y, Matsumura S, Kimura W, Kadowaki M, Higashi T, Tominaga M, Inoue K, Fushiki T. NonPungent Capsaicin Analogs (Capsinoids) Increase Metabolic Rate and Enhance Thermogensis via Gastrointestinal TRPV1 in Mice. Biosci Biotechnol Biochem. 2009 Dec;73(12):2690-2697. Epub 2009 Dec 7.

Kobayashi, Akiko, Osaka, Toshimasa, Namba, Yoshio, Inoue ,Shuji, Lee, Tai Hee, Kimura, Shuichi. Capsaicin activates heat loss and heat production simultaneously and independently in rats. American Journal of Physiology. (1998 July) 275:R92-R98.

Ohnuki, K., Haramizu, S., Oki K., Watanabe, T., Yazawa, S. Admisitration of Capsiate, a Non- Pungent Capsaicin Analog, Promotes Energy Metabolism and Suppresses Body Fat Accumulation in Mice. Biosci Biotechnol Biochem. 2001 Aug;65(12):2735-2740. Epub 2001 Aug 20.

(References- Need a total of 10 sources! Alphabetized and spaced between each source)

Kawabata, F., Inoue, N., Masamoto, Y., Matsumura, S., Kimura, W., Kadowaki, M., Higashi, T., Tominaga, M., Inoue, K., Fushiki, T. 2009. NonPungent Capsaicin Analogs (Capsinoids) Increase Metabolic Rate and Enhance Thermogensis via Gastrointestinal TRPV1 in Mice. Biosci Biotechnol Biochem. 73(12): 2690-2697.

Kobayashi, A., Osaka, T., Namba, Y., Inoue, S., Lee, T., Kimura, S. 1998. Capsaicin activates heat loss and heat production simultaneously and independently in rats. American Journal of Physiology. 275: R92-R98.

Hursel, R., Westerterp-Platenga, M. 2010. Thermogenic ingredients and body weight regulation. International Journal of Obesity. (34): 659-669.

Ohnuki, K., Haramizu, S., Oki, K., Watanabe, T., Yazawa, S. 2001. Admisitration of Capsiate, a Non- Pungent Capsaicin Analog, Promotes Energy Metabolism and Suppresses Body Fat Accumulation in Mice. Biosci Biotechnol Biochem. 65(12): 2735-2740.

Review Form

Department of Biological Sciences

Saddleback College, Mission Viejo, CA 92692

Author (s):______Linda Ko______

Title:___The Effect of Capsaicin on the Metabolic Rate of Male Mus musculus______

Summary

Summarize the paper succinctly and dispassionately. Do not criticize here, just show that you understood the paper.

Capsaicin is a chemical component that triggers the body to undergo thermogenesis. This study tested the metabolic rates of mice when capsaicin was consumed. The hypothesis was that there would be an affect on metabolic rates in Mus musculus. The results showed that there was a difference between the groups tested. However, the hypothesis was rejected because the group without any substances had a suspended basal metabolic rate, cancelling the capsaicin group. Capsaicin did increase the metabolic rates in rats.

General Comments

Generally explain the paper’s strengths and weaknesses and whether they are serious, or important to our current state of knowledge.

The paper’s strength was structure. This paper was not choppy; it flew from one idea to another. The weakness of this paper was grammatical error and some sentence structures need revision. Everything looks good, but the discussion needs more work. Also, discussion needs to be longer (support your results (why?)).

Technical Criticism

Review technical issues, organization and clarity. Provide a table of typographical errors, grammatical errors, and minor textual problems. It's not the reviewer's job to copy Edit the paper, mark the manuscript.

This paper was a final versionThis paper was a rough draft

Organization was good. There were some grammatical errors, but most were sentence structure errors. It is better to have the paper be in third person rather than first or second person.

Recommendation

 This paper should be published as is

 This paper should be published with revision

 This paper should not be published