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The Effects of Rhein and Thymoquinone on Obesity and Diabetes in Diet-Induced Obese Mice." (2015)

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The Effects of Rhein and Thymoquinone on Obesity and Diabetes in Diet-Induced Obese Mice. University of Rhode Island DigitalCommons@URI Senior Honors Projects Honors Program at the University of Rhode Island 2015 The ffecE ts of Rhein and Thymoquinone on Obesity and Diabetes in Diet-induced Obese Mice. Emily Martell University of Rhode ISland, [email protected] Creative Commons License This work is licensed under a Creative Commons Attribution-Noncommercial-Share Alike 3.0 License. Follow this and additional works at: http://digitalcommons.uri.edu/srhonorsprog Part of the Natural Products Chemistry and Pharmacognosy Commons, Other Pharmacy and Pharmaceutical Sciences Commons, and the Pharmaceutics and Drug Design Commons Recommended Citation Martell, Emily, "The Effects of Rhein and Thymoquinone on Obesity and Diabetes in Diet-induced Obese Mice." (2015). Senior Honors Projects. Paper 444. http://digitalcommons.uri.edu/srhonorsprog/444http://digitalcommons.uri.edu/srhonorsprog/444 This Article is brought to you for free and open access by the Honors Program at the University of Rhode Island at DigitalCommons@URI. It has been accepted for inclusion in Senior Honors Projects by an authorized administrator of DigitalCommons@URI. For more information, please contact [email protected]. The effects of Rhein and Thymoquinone on obesity and diabetes in diet-induced obese mice. Emily Martell, Cameron Picard, and Dr. Angela Slitt Department of Biomedical and Pharmaceutical Sciences, College Of Pharmacy University of Rhode Island, Kingston, RI 02881 Introduction Analysis Conclusions Natural product extracts and chemicals isolated from natural products (e.g. plants, berries, seeds) have been commonly used in various types of traditional • There are differences in body weight, FBG, and GTT between the medicines. In addition, some drugs on the market today have been derived from mice feed a HFD and LFD as expected natural product sources. The purpose of our study was to evaluate two natural • The body weights, FBG, and GTT of the groups dosed with rhein products, Rhein and Thymoquinone, as potential anti-diabetic and anti–obesity compared with the groups dosed with the vehicle CO show little agents. According to the Center for Disease Control (CDC), the number of people differences in the US diagnosed with diabetes has increase from 11.9 million people in the year 2000 to 20.8 million people in the year 2011. Rhein is a natural compound and • The body weights and GTT of the groups dosed with TQ a major component of Rheum palmatum, or Rhubarb. It has been used in Chinese compared with the groups dosed with the vehicle CO show little medicine to treat constipation, gastrointestinal hemorrhage, ulcers as well as differences metabolic disorders such as diabetes. Recently, Rhein had been shown to improve • There is a slight difference between the FBG of the HFD group non-alcoholic fatty liver disease (NAFLD) at does of 150 mg/Kg/day in diet induced dosed with TQ compared with the HFD group dosed with the obese mice through reducing body fat, improving serum lipid and glucose metabolism, and decreasing liver lipids, and reversing hepatic steatosis (Sheng et vehicle CO. Which may be an indication that TQ may lower blood al., 2011). Rhein has also been shown to bind to and effect the Liver X receptors sugar but more studies need to be conducted and this group is (LXRs) which play important roles in regulating cholesterol homeostasis, and lipid also missing from the GTT data due to unforeseen circumstances and energy metabolism. (Sheng et al., 2012). Thymoquinone (TQ) is a compound found in the plant Nigella sativa, or black cumin and has been documented to exhibit anti-diabetic, anti-obesity, hypotensive and hypo-lipidemic properties in Figure 1. Changes in average body weights in grams human and animal studies. (Razavi and Hosseinzadeh, 2014). Extracts from throughout the 69 days of dosing within the treatment groups. Nigella sativa significantly increased hepatic and intestinal apolipoprotein A-I which is a major protein component of high density lipoprotein (HDL) secretion. The extract also induced peroxisome proliferator-activated receptor alpha (PPARα) expression by 9-fold and retinoid X receptor alpha (RXRα) expression by 2.5-fold. F a s tin g B lo o d G lu c o s e Future Studies (Haas et al., 2014). The PPARα and RXRα nuclear receptors play and important 3 0 0 role in regulating major lipid metabolism proteins. The purpose of our study was * E to evaluate whether daily administration of Rhein or TQ could improve obesity- S O Figure 2. Fasting induced diabetes in mice. First, male C57BL/6 mice that were 6 weeks of age were C 2 0 0 l U blood glucose • Blood, liver, kidneys, skeletal muscle, adipose tissue, small d L fed a low fat diet (10% kcal, LFD) or a high fat diet (60% kCal, HFD) for 12 weeks. / G g (mg/dL) of different intestine, and colon tissues were collected a further analysis Over the twelve-week period, body weight, fasting blood glucose and glucose m D 1 0 0 tolerance were determined to assess whether the high fat diet could induce a O treatment groups should be conducted in these tissues such as a Glycated O diabetic condition. Starting from week 12, mice were administered canola oil L measured at day 23 hemoglobin (A1c or HbA1c) measurement from the blood tissue B vehicle (CO, 5 ml/kg), Rhein (20 mg/kg, 5 ml/kg in CO), or TQ (1 mg/kg, 5 ml/kg) 0 of dosing. which would identify the average plasma glucose daily. After three weeks of dosing the Rhein and TQ doses were increased to O in Q O in Q concentration over a prolonged period of time. C e T C e T Rhein (50 mg/kg, 2.5 ml/kg) and TQ (10 mg/kg, 2.5 ml/kg). There were six groups of l h l l h l a R a a R a C l C C l C • Further analysis of the tissues after homogenization, which will k a k k a k mice in this study, with the following groups: i) LFD + CO, ii) LFD + Rhein, iii) LFD C C % k % % k % 0 0 0 0 1 % 1 6 % 6 break apart cells so protein expression can be analyzed would be + TQ, iv) HFD + CO, v) HFD + Rhein, vi) HFD + TQ. Body weight and food 0 0 consumption were measured daily. At periodic points throughout the study, 1 6 helpful to determine if the natural products had an effect on T R E A T M E N T fasting blood glucose (FBG) and glucose tolerance (GTT) were measured in the mechanisms of obesity and diabetes. mice. • Western blotting of LXR, PPARα, PPARγ and RXR would be useful to see if Rhein or TQ had an effect on these nuclear Materials and Methods receptor that control energy metabolism. • Western blotting of IRS-1, GLUT2, GLUT4 and SREBP-1c G lu c o s e T o le r a n c e T e s t would be useful to see other mechanisms of glucose and lipid 6 0 0 1 0 % C O regulation by transporters and transcription receptors. E 1 0 % R h e in S O 1 0 % T Q C 4 0 0 ) l U 6 0 % C O d L / g G 6 0 % R h e in m D ( O 2 0 0 O L References B 0 Haas, M.J., Onstead-Haas, L.M., Naem, E., Wong, N.C.W., and Mooradian, A.D. (2014). 0 .0 0 .5 1 .0 1 .5 2 .0 Induction of apolipoprotein A-I gene expression by black seed (Nigella sativa) extracts. Pharm. Biol. 52, 1119–1127. T IM E (H o u rs ) Razavi, B.M., and Hosseinzadeh, H. (2014). A review of the effects of Nigella sativa L. and its constituent, thymoquinone, in metabolic syndrome. J. Endocrinol. Invest. Sheng, X., Wang, M., Lu, M., Xi, B., Sheng, H., and Zang, Y.Q. (2011). Rhein ameliorates fatty liver disease through negative energy balance, hepatic lipogenic Figure 3. GTT done at day 56 of dosing for the treatment regulation, and immunomodulation in diet-induced obese mice. Am. J. groups (Missing 60% TQ group). Physiol. - Endocrinol. Metab. 300, E886–E893. Average blood glucose measurements in (mg/dL) taken at Sheng, X., Zhu, X., Zhang, Y., Cui, G., Peng, L., Lu, X., and Zang, Y.Q. (2012). Rhein protects against obesity and related metabolic disorders through liver X various time points, after giving a oral dose of glucose (1 receptor-mediated uncoupling protein 1 upregulation in brown adipose tissue. mg/kg). Int. J. Biol. Sci. 8, 1375–1384. CDC - Number of Adults - Diagnosed Diabetes - Data & Trends - Diabetes DDT. Poster was presented at 2015 Honors Undergraduate Conference, Kingston, RI After 3 weeks, doses Preformed necropsy Male C57BL/6 mice that were 6 Mice dosed daily by were increased and and collected tissue weeks of age were given a LFD oral gavage continued for 69 days samples or HFD for 12 weeks low fat diet (10% kcal, LFD) Collected: Blood, Liver, Dosed with Canola oil Dosed with CO (2.5 Kidneys, Vehicle (CO, 5 ml/kg), ml/kg), Skeletal muscle, Adipose tissue high fat diet (60% kCal, HFD) Dosed with Rhein Dosed with Rhein (50 Small intestine, (20 mg/kg in 5 ml/kg mg/kg in 2.5 ml/kg CO) And colon CO) Dosed with TQ (1 Dosed with TQ (10 mg/kg, in 5 ml/kg CO mg/kg, in 2.5 ml/kg CO During the 12 week period, body weight, FBG and GTT BodyBody weight weight and and food food consumption consumption were were measured measured daily. were determined to assess daily.At periodic At periodic points points throughout throughout the study, the study, fasting fasting FBG and whether the HFD could FBG and GTT were GTTmeasured were measured induce a diabetic condition Fasting Blood Glucose (FBG) and Glucose Tolerance Testing (GTT) Mice were fasted overnight Average of the two values At 15, 30, 60, and 90 were recorded at initial minutes after the does of blood glucose and FBG glucose, mouse tail blood was collected and The end of the mouse’s tail measured with the was pricked and blood glucometer twice and the was collected and Mice were give a dose of average of the two values measured with a glucose (1g/kg) by oral were recorded.
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