A subscription to JoVE is required to view this content. Sign in or start your free trial.

In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Results
  • Discussion
  • Disclosures
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

Summary

The study investigates the anti-obesity efficacy of Papaver somniferumΒ seeds in obesity-induced Albino rats. Washed poppy seeds effectively reduced weight, lowered glucose levels, and improved lipid profiles without toxicity. In contrast, unwashed seeds altered blood parameters, suggesting potential toxicity and the need for further research.

Abstract

Obesity is a major global health issue, affecting nearly 30% of the population worldwide. Despite the prevalence of obesity, there is currently no data available on the anti-obesity and metabolic effects of Papaver somniferum. The objective of the study was to confirm the anti-obesity and metabolic effects of Papaver somniferum (poppy) seeds in high-fat diet (HFD)-induced obese rats, assessing their impact on weight reduction, lipid profile, and organ toxicity. The experiment was conducted in two phases: a 4 week poppy seed intervention and a 6 week obesity induction trial. Rats were separated into groups and given both washed or unwashed poppy seeds, HFD, and a prescription medication for weight loss. The findings demonstrated that washing poppy seeds significantly decreased weight gain and enhanced lipid profiles, particularly reducing triglycerides, low-density lipoprotein (LDL), and very low-density lipoprotein (VLDL). Additionally, treated groups showed a decrease in glucose levels. However, higher doses of unwashed poppy seeds caused modest liver stress, indicated by raised alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels and renal histopathology showed mild inflammation, but hematological parameters were constant. These results suggest that washed poppy seeds may have the potential to reduce obesity and enhance metabolic health without adverse harm, indicating the need for further research to explore their therapeutic potential.

Introduction

Obesity is a medical condition characterized by an excessively high body fat percentage despite a body mass index (BMI) of less than 201,2. When the body's controlled system fails to maintain the proper balance between nutrients and energy in the body's regulatory system, it results in the accumulation of excess body fat deposited in the body3. The BMI is the primary diagnostic tool used to determine normal weight, overweight, and obesity status. It is commonly used in clinical research to identify individuals with excess weight or obesity. Research indicates that obesity is a disease in and of itself rather than a major cause of any chronic disease4. It is primarily caused by the consumption of excessive calorie-containing food and a sedentary lifestyle5. Genetic weight-gaining mechanisms and prolonged and excessive exposure to food high in energy may also contribute to obesity. The global prevalence of obesity has increased rapidly in recent years, with an estimated 2.1 billion people globally, or 30% of the world's population, suffering from obesity and being overweight. This ratio is predicted to reach 40% by 2030 if the current trends continue to rise. Environmental factors like ecosystem and social issues also influence the development of obesity6,7. Obesity has been linked to several forms of cancer, including uterine, breast, and colon cancer, as well as comorbidities such as dyslipidemia, diabetes, and musculoskeletal disorders (especially osteoarthritis). Furthermore, obesity is also associated with cardiac risk factors, including hyperglycemia and a high body mass index. The most well-known factors that contribute to obesity include hormonal problems and compulsive eating8. While traditional treatments for obesity have not been extensively studied, they generally pose minimal risks. On the other hand, western medications often come with costly side effects that can pose significant health risks. Therefore, an alternate approach to creating safe, efficient anti-obesity medications can be to investigate natural products against obesity. The advancement of phytochemical studies supports the traditional use of therapeutic herbs9, which may be used as an alternative therapy for obesity. Several scientific investigations have shown the effectiveness of herbal medicines in treating obesity for centuries. Earlier studies have demonstrated that medicinal herbs, which contain a variety of pharmacological components, are consumed as food10. There has been discussion about the interest in employing natural herbs as medications. Using these herbs has been associated with very few negative consequences11; these plants can improve digestion and accelerate weight reduction12. Medicinal plants employ a multifaceted approach to address obesity, encompassing five primary mechanisms: appetite suppression, stimulation of thermogenesis and lipid metabolism, inhibition of pancreatic lipase activity, prevention of pathogenesis, and promotion of lipolysis13. Furthermore, natural herbs often contain bioactive compounds that act as digestive enzyme inhibitors, thereby hindering the hydrolysis and absorption of dietary carbohydrates and fats14.

Papaver somniferum, commonly known as the opium poppy or Khashkhash in the subcontinent, is a globally recognized traditional plant with a rich history of use. Various phytochemicals, including alkaloids such as morphine, codeine, noscapine, papaverine, and thebaine, have been isolated from opium15. While the poppy plant is primarily associated with its psychoactive and analgesic properties, its seeds are increasingly recognized for their potential health benefits. Poppy seeds are a rich source of polyunsaturated fatty acids, particularly omega-3 fatty acids, which have been linked to weight management16. Notably, poppy seeds contain Ξ±-linolenic acid, an omega-3 fatty acid whose anti-obesity potential has garnered considerable attention. The 10 and 12 isomers of Ξ±-linolenic acid have been specifically implicated in weight loss. Numerous human studies have demonstrated that supplementation with a combination of these isomers can decrease body fat percentage17. The primary objective of this investigation was to evaluate the effects of P. somniferum on weight reduction in an animal model. Additionally, the study aimed to assess its impact on lipid profile, hematological parameters, kidney and liver function, and adipose tissue histology.

Protocol

All procedures were conducted after the ethical committee of the University of Lahore Pakistan approved them in the meeting held on 21-04-2021 with Registration No: REG. # EPZOOL02193026

1. Housing of animals

  1. House 35 male Wistar albino rats (3 weeks old) individually in standard laboratory cages. Provide ad libitum access to rodent chow and water throughout the experiment.
  2. Maintain the vivarium environment at a constant temperature of 22 Β± 1 Β°C and a relative humidity of 50% Β± 10%. Implement a 12 h:12 h light-dark cycle using artificial illumination.

2. Grouping of animals

  1. At the start of the experiment, divide animals into 2 groups: obese and non-obese. Provide the obese group with a high-fat diet in addition to the normal diet. Feed the non-obese group a normal diet and use it as the control group (control negative) in the experiment.

3. Preparation of high-fat diet (HFD)

  1. Formulate a high-fat diet (HFD) by increasing the proportion of fats derived from both plant and animal sources. Combine the HFD with the standard chow to prepare pellets.
  2. Set the fat content in the HFD at 4 g per 30 g of feed. Place the pellets in the animal cages for 24 h. Weigh and record any remaining pellet after 24 h, then remove it. Provide fresh pellets each day. Maintain this HFD feeding regimen for 6 weeks to induce obesity.
  3. Monitor the body weight of the animals and record every week to check if it is increasing or not. To measure the body weight, put the jar on the weight machine and cover it. Add the rat inside the jar to check the weight.
  4. Supplement the same pellets with poppy seeds for the remaining 4 weeks to investigate their potential weight-loss effects.

4. Grouping of obese animals

  1. After 6 weeks, further divide obese animals into four groups: Group 1 is the obese control (control positive), which continued feeding on HFD but was not provided with any treatment. Group 2 (Standard) is treated with commercially available medicine (see Table of Materials) for obesity control. Group 3 (unwashed) and Group 4 (Washed) are fed with washed and unwashed poppy seeds, respectively.

5. Preparation of poppy seeds

  1. Obtain poppy seeds (P. somniferum) from a local commercial supplier as it is easily available in stores. Sun-dry the seeds for a few hours. Divide the seeds into two groups: unwashed and washed.
  2. Wash the washed group poppy seeds 7x with distilled water, followed by sun drying. Add 500 g of poppy seed to a 1 L beaker, add water and mix with hands. Then discard the water and repeat the procedure 7x to make sure to remove all the impurities, dust, and other particles from the seeds. After the washing, place the seeds in sunlight to dry.

6. Calculation of dosage and preparation of feed

  1. Determine the dosage based on the animal's feed intake, standardized at 30 g per individual. Administer poppy seeds at a concentration of 0.5 g per 30 g of feed.
  2. Incorporate the poppy seeds into the feed by mixing them with distilled water to form a pellet. Prepare the pellets fresh daily throughout the experimental period of 4 weeks.

7. Dissection of rats and blood collection

  1. Fast the rats for 24 h. Sacrifice the animal by using chloroform and collect the blood and organ samples for further procedures as described below.
  2. Anesthetize the rats using chloroform. Administer chloroform by a trained lab technician at the rate of 1% inhaled chloroform (0.05 mL/L) and ensure the rat before is properly anesthetized by pinching the foot of the rat.
  3. Perform dissection using standard dissecting box instruments. Disinfect a sharp knife, scissors, and forceps prior to starting the dissection. During dissection remove kidney, liver, and adipose tissue samples. Preserve the excised samples in formalin-filled centrifuge tubes.
  4. Collect blood samples in EDTA-coated vials. Extract 12 mL of blood from each animal and then further divide 3 mL each to test kidney function, liver functioning test, and lipid profiling.
  5. Centrifuge the collected blood samples at 1,957 x g for 5 min. Separate blood serum from the rest as a clear or yellowish liquid. Separate the serum and aliquot it into micro centrifuge tubes for further biochemical analyses.

8. Sample analysis

  1. Determine the total lipid profile, including total cholesterol, HDL-cholesterol, LDL-cholesterol, triglycerides, and VLDL, using standard enzymatic colorimetric methods18. Base the determination of HDL cholesterol on a time-consuming precipitation procedure18.
  2. Monitor rat glucose levels regularly during the whole experiment using a blood glucose monitor GL-11019.

9. Evaluation of toxicity

  1. Assess the toxicological impact of Papaver somniferum on the parameters ALT, AST, ALP, TP, and bilirubin using liver function tests20. Measure serum creatinine levels following the methodology outlined in 21.
  2. Determine hematological parameters, including red blood cell (RBC) and white blood cell (WBC) counts, using a Neubauer hemocytometer. Assess hemoglobin concentration using the cyanmethemoglobin method.
  3. Conduct a histopathological examination of the collected tissue samples. Stain sections with hematoxylin and eosin (H&E) for microscopic evaluation.

10. Statistical analysis

  1. Use SPSS software version 16 for statistical analysis. Apply an independent t-test to the first month's data for the obesity induction trial.
  2. Utilize repeated measures ANOVA with LSD post-hoc test for the second phase. Employ one-way ANOVA with LSD to analyze blood parameters for toxicity assessment
  3. Define statistical significance as p ≀ 0.05, with p ≀ 0.001 considered highly significant.

Results

Rats with an initial body weight of 40-45 g (weaning stage) were selected for the 70 day experiment, which was divided into two phases. The first phase, lasting 6 weeks, involved inducing obesity in the experimental group by supplementing their standard chow with a high-fat diet (HFD; Figure 1). In the subsequent 4 week phase, the obese rats were administered Papaver somniferum. Body weight was measured at the beginning and end of the experiment. Repeated measures A...

Discussion

The protocol has the following critical steps to be taken care of. The first critical step was to induce obesity in the animals by using High fat diet. The second step was to monitor animals which are on high fat diet that they do not develop diabetes or any other disease due to HFD.

The protocol also has the following limitations. Due to academic restrictions, the experimental time to study the exact effect of poppy seeds was only 10 weeks. A more detailed study of poppy seeds with different ...

Disclosures

The authors have nothing to disclose.

Acknowledgements

The authors would like to extend their sincere appreciation to the Researchers Supporting Project Number (RSPD2025R986), King Saud University, Riyadh, Saudi Arabia.

Materials

NameCompanyCatalog NumberComments
Blood glucose monitor GL-110CertezaGL-110A portable device used to measure blood glucose levels.
CentrifugeEppendorf22620005A laboratory equipment that spins samples at high speeds to separate components based on density.
EDTA-coated vialsBD Vacutainer367861Tubes used for collecting blood samples, preventing clotting by binding calcium ions.
Eppendorf tubesEppendorf0030 120.094Small plastic tubes used for storing and handling small volumes of liquid.
Falcon tubesCorning352059Conical tubes used for centrifugation and various laboratory applications.
MicroscopeOlympusCX23RF100An optical instrument used for viewing small objects, typically magnified through lenses.
Neubauer hemocytometerHawksleyH.H1A specialized slide used for counting cells under a microscope.
Orlistat120 mgWindlas Biotech Ltd
Pipette tipsEppendorf0030 073.435Disposable tips used with pipettes for transferring liquids accurately.
Serological pipettesFalcon357551Graduated pipettes used for transferring liquids in larger volumes.
SPSS software version 16IBMN/A (software, not a physical product)A statistical software package used for data analysis.
Standard laboratory cage (Super Mouse 750)Lab Products, Inc.10021A cage designed to house laboratory mice, providing a controlled environment.

References

  1. Da Costa, G. F. et al. The beneficial effect of anthocyanidin-rich Vitis vinifera l. Grape skin extract on metabolic changes induced by a high-fat diet in mice involves anti-inflammatory and antioxidant actions. Phytother Res. 31 (10), 1621-1632 (2017).
  2. Andersen, M. M., Varga, S., Folker, A. P. On the definition of stigma. J Evaluat Clin Pract. 28 (5), 847-853 (2022).
  3. Archer, E. Hill, J. O. Body, and fat mass are not regulated, controlled, or defended: An introduction to the invisible hand and 'competition ' models of metabolism. Progr Cardiovasc Dis. 79, 56-64 (2023).
  4. Imhagen, A., Karlsson, J., Jansson, S., AnderzΓ©n-Carlsson, A. A lifelong struggle for a lighter tomorrow: A qualitative study on experiences of obesity in primary healthcare patients. J Clin Nurs. 32 (5-6), 834-846 (2023).
  5. Alonso-Castro, A. J. et al. Self-treatment with herbal products for weight loss among overweight and obese subjects from central Mexico. J Ethnopharmacol. 234, 21-26 (2019).
  6. Alsareii, S. A. et al. Iot framework for a decision-making system of obesity and overweight extrapolation among children, youths, and adults. Life. 12 (9), 1414 (2022).
  7. Bautista, R. J. H., Mahmoud, A. M., KΓΆnigsberg, M., Guerrero, N. E. L. D. Obesity: Pathophysiology, monosodium glutamate-induced model and anti-obesity medicinal plants. Biomed Pharmacother. 111, 503-516 (2019).
  8. Radin, R. M. et al. Do stress eating or compulsive eating influence metabolic health in a mindfulness-based weight loss intervention? Health Psychol. 39 (2), 147 (2020).
  9. De Freitas Junior, L. M. De Almeida Jr, E. B. Medicinal plants for the treatment of obesity: Ethnopharmacological approach and chemical and biological studies. Am J Transl Res. 9 (5), 2050 (2017).
  10. Awuchi, C. G. Medicinal plants: The medical, food, and nutritional biochemistry and uses. Int J Adv Acad Res. 5 (11), 220-241 (2019).
  11. Ozioma, E. O. J. Chinwe, O. a. N. Herbal medicines in african traditional medicine. Herbal Med. 10, 191-214 (2019).
  12. Al-Snafi, A. E., Singh, S., Bhatt, P., Kumar, V. A review on prescription and non-prescription appetite suppressants and evidence-based method to treat overweight and obesity. GSC Biol Pharmaceut Sci. 19 (3), 148-155 (2022).
  13. Saad, B., Zaid, H., Shanak, S., Kadan, S. Anti-diabetes and Anti-obesity Medicinal Plants and Phytochemicals: Safety, Efficacy, and Action Mechanisms. Springer Cham (2017).
  14. Ardeshirlarijani, E. et al. Potential anti-obesity effects of some medicinal herbs: In vitro Ξ±-amylase, Ξ±-glucosidase, and lipase inhibitory activity. Int Biol Biomed J. 5 (2), 1-8 (2019).
  15. Haber, I., Pergolizzi, J., Lequang, J. A. Poppy seed tea: A short review and case study. Pain Ther. 8, 151-155 (2019).
  16. Liu, R. et al. Omega-3 polyunsaturated fatty acids prevent obesity by improving tricarboxylic acid cycle homeostasis. J Nutri Biochem. 88, 108503 (2021).
  17. Basak, S. Duttaroy, A. K. Conjugated linoleic acid and its beneficial effects in obesity, cardiovascular disease, and cancer. Nutrients. 12 (7), 1913 (2020).
  18. Mcclatchey, K. Clinical laboratory medicine. Lippincott Williams & Wilkins, Philadelphia, USA (2002).
  19. Padmaja, T. K., Naidu, P. B., Kumar, G. E. N. H., Ganapathy, S., Balaji, M. Antiobesity activity of bauhinia purpurea extract: Effect on hormones and lipid profile in high-calorie diet-induced obese rats. Adv Biosci Biotechnol. 5 (11), 861 (2014).
  20. BΓΌlbΓΌl, T., GΓΌr, E., Bozkurt, F., Eryavuz, A., BΓΌlbΓΌl, A. Biochemical, hematological and histopathological evaluation of the food-safety of the leaf extract of Papaver somniferum in rats. J Trad Compl Med. 10 (6), 544-554 (2021).
  21. Precious, I. O., Ayoka, T. O., Nnadi, C. O. In-vivo sub-chronic toxicological evaluation of extract of vernonia glaberrima leaves in experimental rats. Notulae Sci Biol. 14 (2), 11181-11181 (2022).
  22. Bonizzi, A., Piuri, G., Corsi, F., Cazzola, R., Mazzucchelli, S. Hdl dysfunctionality: Clinical relevance of quality rather than quantity. Biomedicines. 9 (7), 729 (2021).
  23. Cabot, S. Hepatitis and aids: A plan to recover with complementary and modern treatments. SCB International (2015).
  24. Mohamed, S. S. Fayed, A. H. M. Anti-obesity synergistic effect of pomegranate seed oil (pso) and arabic gum (ag) in albino rats. Int J Vet Sci. 9 (1), 84-89 (2020).
  25. Czaja, A. J. Hepatic inflammation and progressive liver fibrosis in chronic liver disease. World J Gastroenterol. 20 (10), 2515 (2014).
  26. Barrett, A. H., Farhadi, N. F., Smith, T. J. Slowing starch digestion and inhibiting digestive enzyme activity using plant flavanols/tannins-a review of efficacy and mechanisms. Lwt. 87, 394-399 (2018).
  27. Nakajima, K., Muneyuki, T., Munakata, H., Kakei, M. Revisiting the cardiometabolic relevance of serum amylase. BMC Res Notes. 4, 1-5 (2011).
  28. Lesmana, R. et al. Nutmeg extract potentially alters the characteristics of white adipose tissue in rats. Vet Med Sci. 7 (2), 512-520 (2021).
  29. Azman, K. F. et al. Antiobesity effect of tamarindus indica l. Pulp aqueous extract in high-fat diet-induced obese rats. J Natural Med. 66, 333-342 (2012).

Reprints and Permissions

Request permission to reuse the text or figures of this JoVE article

Request Permission

Explore More Articles

Papaver SomniferumAnti obesityObesity ModelMetabolic EffectsHigh fat DietWeight ReductionLipid ProfilePoppy Seed InterventionTriglyceridesLDLVLDLGlucose LevelsLiver StressALTASTRenal HistopathologyTherapeutic Potential

This article has been published

Video Coming Soon

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright Β© 2025 MyJoVE Corporation. All rights reserved