The Ginseng Enigma: Regulating the Symptoms of Diabetes Mellitus Using Panax Quinquefolius

Transcription

1 The Ginseng Enigma: Regulating the Symptoms of Diabetes Mellitus Using Panax Quinquefolius Tran Luong Southmoore High School 2901 S. Santa Fe Moore, OK

2 Table of Contents Abstract 3 Introduction 4 Questions and Hypothesis 7 Procedures 7 Results 9 Conclusion 16 Bibliography 18 Acknowledgements 20 2

3 Abstract This project was conducted to look at the effects of Panax quinquefolius ( and if its components are successful in creating positive outcomes on diabetic mellitus symptoms, such as reduced mental and physical abilities and elevated blood sugar levels. In this study, ten diabetic mice and ten control mice were used for the comparison of their blood sugar levels, weight, and mental and physical abilities. The mice were then divided into four groups of five mice. They are then fed the Purina high fat diet and 0.05 grams of both P. quinquefolius and non-p. quinquefolius (butter) dosages. Group A and Group C consisted of mice that were Panax quinquefolius treated, but the mice in Group A were diabetic while the mice in Group C were control. Group B and Group D had mice that were not P. quinquefolius treated, but Group B consisted of diabetic mice while Group D consisted of control. Their blood was then tested twice a week. Afterwards, they were then put through the Morris Water Maze. The data from this experiment supported that the consumption of P. quinquefolius does have a positive effect on diabetic and non-diabetic organisms and that it not only helps lower and maintain the blood sugar levels and weight in diabetic and non-diabetic organisms but also help improve their mental and physical abilities. 3

4 Introduction According to the National Diabetes Information Clearinghouse s article Diabetes Overview (2008), diabetes mellitus is a metabolism disorder disease in which the body cannot properly store and use the natural sugar, glucose. This results in abnormally high blood sugar. Glucose is taken in through the consumption of foods and, once digested, is used as energy by the body s cells. However, in order for the cells to take in the glucose, a sufficient amount of insulin must be present. Insulin is a hormone produced by the pancreas (Gerdes, 2003). In diabetes mellitus, this gland behind the stomach is unable to produce the appropriate amount of insulin or the cells are unable to use the insulin properly to convert the glucose into energy. There are three main types of diabetes mellitus: Type 1, Type 2, and gestational. In type 1 diabetes mellitus, the immune system attacks and then destroys the beta cells, which are located in the pancreas, that produce insulin. People who have type 1 diabetes mellitus must take insulin medication to live. In type 2 diabetes mellitus, the pancreas produces enough insulin. However, due to insulin resistance, the body cannot use the insulin effectively. Insulin resistance can be caused by obesity and stress. Gestational diabetes mellitus develop in women who are late in their pregnancy. Although this form of diabetes usually disappears after the birth of the baby, women who have had gestational diabetes have a 40 to 60 percent chance of developing type 2 diabetes within 5 to 10 years. (NDIC, 2008). Panax quinquefolius ( is an ancient root that has been used as a drug for humans for thousands of years, dating back to ancient Asia. Its components are separated into ginsenoside Rb1 and ginsenoside Rh2. According to (Xie, et al. 2004) and (Shang, et al. 2004) Ginsenoside Rb1 is claimed to stimulate glucose uptake through insulin, and ginsenoside Rh2 is 4

5 suspected to increase the secretion of insulin. These claims are from holistic medicine but not scientifically proven. According to (Craker, 2011), P. quinquefolius isn t just said to lower blood sugar in diabetic organisms but nondiabetic ones, as well. Also, studies have shown that more men are affected with diabetes mellitus and their symptoms are much more visible. This study will focus on whether herbal supplements, such as P. quinquefolius, will have a positive effect on the symptoms of diabetes mellitus. With diabetes mellitus comes not only the hassle of glucose test meters and strips, a careful watch of a daily diet and exercise, and the side effects caused by the many diabetes mellitus prescribed medications, but also the possibility of obesity, blindness, high blood pressure, and high blood sugar. Not only will those medications result in short term and long term side effects, but they will also cost an immense amount of money. If the data supports using P. quinquefolius to alleviate symptoms of diabetes, then adding it alongside with the medications would conceivably lessen the amount of medications needed to be taken which in turn could result in fewer side effects and less money spent. In this study, P. quinquefolius will be used as a substitute for diabetic medication, and an appropriate mouse model was chosen as a substitute for humans. According to (Craker 2011) P. quinquefolius has been used for many centuries by the Chinese and is said to not only lower blood pressure but blood sugar, as well. Because of this reason, this research is being conducted to determine if P. quinquefolius can be a holistic alternative to diabetic medicine. Mice, like humans, are vertebrates and have very similar body systems. However, with mice, since they live a much shorter time span than humans do, effects of the P. quinquefolius maybe measureable as a reduction in blood glucose in a much shorter period of time than with a human. 5

6 The mice that were used in this experiment came from Jackson s Laboratory, a professional laboratory that engineers and sells, under a license, genetically engineered mice for scientific research. They were all male mice with the strain C57BL/6J at six weeks of age. The control mice were of the same strain but lack the genetically engineered diabetes gene. The diet that the mice will be consuming for the first 8 weeks is a high fat Western diet from Purina. This diet was used to replicate the diet of the westernized country in the world. Because of the world s accelerated modernization, the human population is slowly shifting towards a more Western diet and lifestyle. The intention of using the high fat Western diet was to mimic the modern human of many countries around the world. While a normal, healthy diet contains only about 11% fat, the Purina high fat Western diet contains about 58% fat. These two diets were used to compare the blood glucose levels between a high fat Western diet and a normal, healthy diet. Thirty years ago, a scientist by the name of Richard Morris designed the Morris Water Maze. This test, according to (D Hooge 2001) is used to test the mental abilities, such as thinking, learning, and memory, of lab mice. Each mouse performed the Morris Water Maze in this research. The spatial learning and memory were acquired through repeated trials. This allowed the mice to realize where the platform is located and tested their ability to remember where it was and return back to it, once placed back in the water again. In this test, each mouse was placed on a clear plastic platform, that was placed in water, and then dropped in the water and timed to observe their mental and physical abilities that was affected by diabetic symptoms and P. quinquefolius. 6

7 Questions and Hypotheses What are the measurable effects of ginsenoside in Panax quinquefolius on diabetic symptoms? If the herbal supplement Panax quinquefolius is given to a diabetic organism, then measurable improvements can be noted in diabetes mellitus symptoms, by decreasing blood sugar levels and body weight, and improving mental and physical abilities. What are the measurable effects of ginsenoside in Panax quinquefolius on non-diabetic organisms? If the herbal supplement Panax quinquefolius is given to a nondiabetic organism, then measurable improvements can be noted in the organism s blood sugar, body weight, and mental and physical abilities. Procedures The mice were divided into four groups with five mice each. The groups were male diabetic mice treated with P. quinquefolius (Group A), male diabetic mice not treated with P. quinquefolius (Group B), control male mice treated with P. quinquefolius (Group C), and control male mice not given P. quinquefolius (Group D). Each mouse were fed one pellet of Purina High Fat Western Diet (58% fat) a day for eight weeks. Starting at week 9, all mice were then switched over to a normal diet (11% fat). A measurement gram of P. quinquefolius extraction medication was then mixed with grams of melted control butter, for easy ingestion by the mice. Every day at 12:30 P.M., mice in Group A and Group C were fed a dosage of 0.05 grams of P. quinquefolius extract 7

8 medication, and mice in Group B and Group D were fed a piece of 0.05 grams of control, nontreated butter. Every mouse had their blood tested every Tuesday and Friday of every week at 3:00 P.M. The end of each mouse s tail was gently scrapped until a slight trace of blood was visible. The meter that was used was an Alpha Trak meter with #7 glucose strips, made especially for the use of testing animal blood. The process was then repeated for every mouse, and the data was then recorded. The mice were then weighed every Friday at 3:00 P.M. The Morris Water Maze was used for measuring physical agility and mental awareness in the mice. A clear platform was placed in the center of the bucket of water (water was mixed with white tempera paint for camouflage). Each mouse was then placed on that platform for 15 seconds and then placed down on the ground for 10 seconds. After that, each mouse was dropped in the same area in the bucket for them to find the platform again. Once the platform was found, the time was recorded. 8

9 Figure 1 Blood Glucose Levels (mg/dl) Results Average Blood Glucose Levels in Diabetic and Control Mice on a High Fat Western Diet Over an Eight Week Period Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7 Week 8 Group A (diabetic, ginseng treated) Group B (diabetic, no Group C (control, ginseng treated) Group D (control, no Time (weeks) As seen in the results, Group A (P. quinquefolius treated) for the first two weeks had a higher average blood glucose level than Group B (non-p. quinquefolius). However, numerically, from the third week on, the average blood glucose level of Group A was significantly much lower than that of Group B s. Group C s (control, P. quinquefolius treated) average blood glucose compared to Group D s (control, non-p. quinquefolius) was usually significantly higher, but stable. Figure 2 Blood Glucose Levels (mg/dl) Blood Glucose Levels in Diabetic and Control Mice on a High Fat Western Diet Over an Eight Week Period Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7 Week 8 Time (weeks) Group A (diabetic, ginseng treated) Group B (diabetic, no Group C (control, ginseng treated) Group D (control, no 9

10 Figure 3 Weight (grams) Average Weight of Diabetic and Control Mice on a High Fat Western Diet Over an Eight Week Period Group A (diabetic, ginseng treated) Group B (diabetic, no Group C (control, ginseng treated) 5 0 Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7 Week 8 Time (weeks) Group D (control, no Group A s and Group B s weight were about the same during week 1 and 2, but, numerically, Group B s (diabetic, non-p. quinquefolius) average weight then significantly rose and remained higher than Group A s (diabetic, P. quinquefolius treated). Both Group A s and Group B s weight rose linearly the same. Group D s (control, non-p. quinquefolius) remained constantly higher than Group C s (control, P. quinquefolius treated).. Figure 4 Weight (grams) Average Weight of Diabetic and Control Mice on a High Fat Western Diet Over an Eight Week Period Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7 Week 8 Time (weeks) Group A (diabetic, ginseng treated) Group B (diabetic, no Group C (control, ginseng treated) Group D (control, no 10

11 Figure Average Blood Glucose Levels in Diabetic and Controlled Mice on a Normal Diet over a Two Week Period Blood Glucose Level (mg/dl) As seen from the graph, each group s average blood glucose levels have decreased since they switched over to a normal diet. However, Group B s (diabetic, non- P. quinquefolius) average blood glucose level remained much higher than that of Group A s (diabetic, P. quinquefolius). Group C s (control, P. quinquefolius treated) blood glucose level is also lower than that of Group D s (control, non-p. quinquefolius). Figure 6 Weight (grams) Week 9 Week 10 Time (weeks) Average Weight of Diabetic and Control Mice on a Normal Diet Over a Period of Two Weeks Week 9 Week 10 Time (weeks) Group A (diabetic, ginseng treated) Group B (diabetic, no Group C (control, ginseng treated) Group D (control, no Group A (diabetic, ginseng treated) Group B (diabetic, no Group C (control, ginseng treated) Group D (control, no According to the graph each group s average weight has significantly decreased since the switch to a normal diet. Group B s (diabetic, non-p. quinquefolius) average weight is still higher than that of Group A s (diabetic, P. quinquefolius), while Group C s (control, P. quinquefolius) average weight is now higher than that of Group D s (control, non-p. quinquefolius). 11

12 Figure Average Glucose Level of Group A on a High Fat Western Diet Over an Eight Week Period Blood glucose level (mg/dl) Group A (diabetic, ginseng treated) 0 Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7 Week 8 Time (weeks) With the Panax quinquefolius dosage, the average glucose level of the mice in Group A decreased after two weeks of being treated. It then remains fairly constant. Figure Average Blood Glucose Level of Group B on a High Fat Western Diet Over an Eight Week Period Blood glucose level (mg/dl) Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7 Week 8 Group B (diabetic, no ginseng ) Time (weeks) With no Panax quinquefolius, Group B s average blood glucose level remains high. Around the time of week 5, the group s blood glucose level begins to increase 12

13 Figure 9 Blood Glucose Level (mg/dl) Group C s average blood glucose level increased after the first week, levels off, then remained constant afterwards for the rest of the time Figure 10 Average Blood Glucose Level of Group C on a High Fat Western Diet Over an Eight Week Period Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7 Week 8 Time (weeks) Group C (control, ginseng treated) Blood Glucose Level (mg/dl) Average Blood Glucose Level of Group D on a High Fat Wester Diet Over an Eight Week Period Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7 Week 8 Group D (control, no Time (weeks) Group D s average blood glucose levels is very fluctuating. Overall, it decreases, then increases, then resumes to decrease. It has no definite rate or leveling. 13

14 Figure 11 Time (seconds) Average Time of Diabetic and Control Mice on the Morris Water Maze Group A (diabetic, ginseng treated) Group B (diabetic, no Group C (control, Group D (control, no 10 0 Test 1 Test 2 Test 3 Test 4 Test 5 Test Number Group A and Group C (P. quinquefolius treated) had lower time averages in the Morris Water Maze test than did Group B and D (non-p. quinquefolius). Group A had the lowest while Group D had the overall highest times. Discussion of Results As seen in the results of Figure 1, the diabetic P. quinquefolius treated group for the first two weeks had a higher average blood glucose level than the diabetic non-p. quinquefolius group. However, from the third week on, the average blood glucose level of the diabetic P. quinquefolius treated group was significantly lower than that of the diabetic non-p. quinquefolius group. As seen in the results of Figure 3, the diabetic P. quinquefolius treated group s and the diabetic non-p. quinquefolius group s weight were about the same during week 1 and 2, but the diabetic non-p. quinquefolius group s average weight then significantly rose higher than the diabetic P. quinquefolius treated group. Both the diabetic P. quinquefolius treated groups and the diabetic non-p. quinquefolius group s weight increased linearly the same. The control P. quinquefolius treated group had a more stable blood glucose and a lower weight average than the control non-p. quinquefolius group. Looking at both Figure 5 and Figure 6, it can be seen that 14

15 once the groups diet were switched to a normal diet, the diabetic P. quinquefolius treated group s average blood glucose levels and average weight still remained lower than that of the diabetic non-p. quinquefolius group. Referring back again to Figure 5 and 6, the control P. quinquefolius treated group had a lower blood glucose level but a slightly higher weight average than the control non-p. quinquefolius. Looking at the average blood glucose for each group individually, the diabetic P. quinquefolius treated group s average blood glucose levels (Figure 7) had decreased after a week of being on the P. quinquefolius dosage. Afterwards, it remained constant at a lower level than the diabetic non-p. quinquefolius group s average blood glucose level. The diabetic non-p. quinquefolius group s average blood glucose level (Figure 8) remained high throughout the entire experiment. During the last few weeks of experimenting, their blood glucose level began to increase. For the control P. quinquefolius treated group (Figure 9), its average blood glucose level increased after the first week, levels off, then remained constant afterwards for the rest of the time. The control non-p. quinquefolius group s average blood glucose (Figure 10) is somewhat similar to the control P. quinquefolius treated group s but its levels are more fluctuating. Overall, it decreases, then increases, and then resumes decreasing. It has no definite rate or leveling. With the Morris Water Maze, Group A (diabetic, P. quinquefolius treated) had a much lower times than Group B (diabetic, non-p. quinquefoliu) did, Group C (control, P.quinquefolius treated) also had the lower times compared to Group D (control, P. quinquefolius). With diabetic mice, their physical and mental abilities are much lower than that of the controls. However, Group A was much faster in remembering and finding the platform than Group B was. Once placed in the water, most of the mice in Group A instantly turned to the platform s direction and 15

16 swam toward it, while Group B had a harder time remembering where the platform was. The control groups had much higher energy levels than the diabetic groups, therefore they spent a longer time swimming around, trying to find an escape route. However, Group C s memory levels were higher than that of Group D s, so their times to find the platform, once they were tired, were much lower. Conclusion Hypothesis 1 is supported by Figure 1, 3, 5, 6, 7, and 8. It can be seen that compared to the group of diabetic mice that were not give the P. quinquefolius treatment, the group of diabetic mice that were given the P. quinquefolius dosages had overall lower blood glucose levels and weight. The data supports that P. quinquefolius does have a positive effect on diabetic symptoms. Hypothesis 2 is supported, as seen from Figure 1, 3, 5, 6, 9, and 10. The group of control mice that were given the P. quinquefolius treatment had very similar, if not slightly higher, blood glucose levels and lower weight than the group of control mice that were not given the P. quinquefolius treatment. However, their performance levels in the Morris Water Maze were much higher. They were quicker to learn and memorize where the platform was. One of the main reasons why these hypotheses are supported is due to the P-value. The P-value for this experiment is , which means that this project contains % certainty that results are produced by the ginseng P. quinquefolius supplement. If given another chance to perform this project again, changes will be needed to be added for improvement. The mice will be ordered at an age of 8 weeks, rather than at 6 weeks. This is to provide more time for the diabetic symptoms in the mice to be more visible. Every time the mice are tested, taking their blood and performing the Morris Water Maze, they must be tested 16

17 all at once. This is to prevent their stress levels from rising, which will affect not only their blood glucose levels but also their physical abilities. It was noted that the mice seemed to detect that they were going to be tested, so the mice that were tested later had higher stress levels, that could have contributed to higher blood glucose levels. Some errors that were noted in the experiment are that the mice sometimes were not tested at the exact same time for every test. This is the same for when the mice were given their dosages, as well. Another margin of error is that some mice ate more of their high fat diet food and dosages than others. The time that the mice ate their high fat diet food and dosages has an effect on their blood glucose levels and physical abilities, as well. Some mice ate it right was it was given while some waited longer to consume it. Diabetes mellitus has not only been a problem in the United States for the past few decades, but it has also reached to the developed countries around the world that have started to embrace the Western diets and way of life. Problems such as high blood sugar levels, chances of obesity/weight gain, and reduced physical abilities accompany diabetes mellitus. The hassles of purchasing and consuming diabetes mellitus medication bring about not only expensive medication but also side effects. This study shows and supports that P. quinquefolius has a positive effect on diabetic symptoms and this researcher recommends that P. quinquefolius be used as a tool to help control diabetes mellitus in humans. Further research is recommended as to P. quinquefolius s effect on human diabetes mellitus. If P. quinquefolius can help lower and keep in balance the blood sugar levels and body weight and increase the mental awareness and physical abilities in people with diabetes mellitus, then taking the supplement alongside with a diabetes medication could be very beneficial. 17

18 Bibliography Craker, Lyle E. "Ginseng." World Book Student. World Book, Web. 23 Sept D'Hooge, R. "Applications of the Morris Water Maze in the Study of Learning and Memory." PubMed.gov. NCBI, Aug Web. 12 Dec < Harlan, David. "Diagnosis of Diabetes - National Diabetes Information Clearinghouse." National Diabetes Information Clearinghouse Home. Clearinghouse, Oct Web. 22 Sept < "Diabetes Overview - National Diabetes Information Clearinghouse." National Diabetes Information Clearinghouse Home. Clearinghouse, Nov Web. 27 Sept < Brown, David. "Global Diabetes Rate Soars." Washington Post. 26 Jun 2011: A.1. SIRS Researcher. Web. 27 Sep "Anti-diabetic Effect of Ginsenoside Re in Ob/ob Mice." NCBI. PubMed, 10 June Web. 20 Sept < "Ginseng and Ginsenoside Re Do Not Improve β-cell Function or Insulin Sensitivity in Overweight and Obese Subjects with Impaired Glucose Tolerance or Diabetes."NCBI. PubMed, 16 Mar Web. 20 Sept < Morholt, Evelyn, and Paul Brandwein. A Sourcebook for the Biological Sciences. Orlando, Florida: Saunders College Publishers,

19 Gerdes, Louise I., ed. Diabetes. Greenhaven, Print. Shang, W., Y. Yang, L. Zhou, B. Jiang, H. Jin, and M. Chen. PubMed.gov. NCBI, 11 June Web. 20 Sept Silverstein, Alvin, Virginia B. Silverstein, and Laura Silverstein. Nunn. The Diabetes Update. Berkeley Heights, NJ: Enslow, Print. 19

20 Acknowledgements I would like to take the time now to just thank a few very kind and special people who have been of great help to me throughout the time of my project. I d like to thank all the companies who donated my supplies and equipment. Aparo Maya donated the AlphaTRAK glucose meter and glucose strips. Purina donated their high fat Western diet for mice. I would also like to thank Mrs. Wyatt, my wonderful mentor who has never failed to help me or solve any problems that I had, and Mrs. Marsh, for always providing me new questions for further research. I also like to thank some of my kind and wonderful classmates for taking time out to help me out with my project: Blake Allen, Melissa Zanabria, Kyle Wallace, and Joann Lee. And, of course, a special Thank You to my wonderful parents who have been so supportive and helpful with everything. 20