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247Abdulaziz K. Al Mouslem
Department of Pharmaceutical Sciences, College of Clinical Pharmacy, King Faisal University, Al Ahsa 31982, Saudi Arabia. Corresponding Author E-mail: aalmoslem@kfu.edu.sa
DOI : https://dx.doi.org/10.13005/bpj/2362
Abstract
Objectives: To investigate the hypoglycemic and hypolipidemic potentials of Picris babylonica extract in streptozotocin (STZ) induced diabetes in rats. Methods: Animals were injected with 40mg/kg of STZ to induce diabetes, a common diabetic model. Development of the disease were assessed by measuring blood glucose level 3 days prior systemic administration of STZ and following STZ injection. Animals received 200mg/kg and 400mg/kg of Picris babylonica extract and 0.6mg/kg of glibenclamide, standard, by oral rout for 14 consecutive days. Results: Administration of the Picris babylonica extract significantly decreased serum blood glucose, total cholesterol, triglycerides, and low-density lipoprotein-cholesterol. In addition, high density lipoprotein-cholesterol level significantly enhanced as compared to standard. Conclusion: Picris babylonica extract demonstrated beneficial effects in lowering blood glucose and improving lipid profile, therefore, Picris babylonica extract could be developed as hypoglycemic and hypolipidemic therapy.
Keywords
Asteraceae; DPPH; Hypolipidemic; Streptozotocin Antidiabetic; Picris babylonica
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| Copy the following to cite this article: Al-Mouslem A. K. Antidiabetic and Hypolipidemic Potentials of Extract of Picris Babylonica in Streptozotocin-Induced Diabetic Model in Rats. Biomed Pharmacol J 2022;15(1). |
| Copy the following to cite this URL: Al-Mouslem A. K. Antidiabetic and Hypolipidemic Potentials of Extract of Picris Babylonica in Streptozotocin-Induced Diabetic Model in Rats. Biomed Pharmacol J 2022;15(1). Available from: https://bit.ly/3LbvsGu |
Introduction
One of the challenging health concerns is Diabetes mellitus (DM), which affects approximately 537 million adults globally with an estimated 6.7 million deaths in 2021. The number of incidence is anticipated to increase up to 643 million in 2030 and 784 million in 2045. 1. DM is a metabolic disorder occasioned by abnormally increased-blood sugar level (hyperglycemia). In addition, it is very often correlated with disturbances in metabolism of carbohydrate, protein, and lipid2, 3. These metabolic disturbances are triggered by either insulin deficiency and/or insulin resistance 4. It is well-known that there is an association between hyperglycemia and diabetic dyslipidemia, which leads to numerous complications including microvascular damages and functional impairments of organs such as the heart, the kidney, and central nervous system 3, 5.
Large body of evidence indicated that type 2 diabetes mellitus represents greater than 90% of all cases 1. Usually, pharmacological interventions are required to control blood glucose level in most of these cases. However, the most used antidiabetic agents have limited efficacy and serious side effects such as hypoglycemia and diabetic keto-acidosis 6, 7. Therefore, identifying effective antidiabetic and hypolipidemic agents with fewer side effects are warranted. Furthermore, tremendous efforts have been utilized on finding an alternative medicine from naturally existing products such as herbal remedies due to their enhanced efficacy and limited undesirable effects 8, 9.
Compositae, also known as Asteraceae, is considered as one of the largest families of flowering plants and comprises around 22,000 species, distributed into 1620 genera. Some of these genera were reported to have an antidiabetic activity 10, 11. Among these genera, Picris known as an important genus comprises around 40 species12, 13. Many plants of genus Picris were investigated previously and reported to contain different types of biologically active constituents like, phenolic constituents, flavonoids, and various types of terpenoids 14, 15. Some species of Picris were found to have an antidiabetic activity 16. Picris babylonica (P. babylonica) spreads in many parts of Saudi Arabia, including Riyadh region. According to the recent reports, P. babylonica was investigated for its volatile constituents and its phytochemical contents for various plant parts 12, 15. These findings encourage us to carry out this experimental work, in which we explore the antidiabetic and hypolipidemic potentials of P. babylonica in diabetes induced rats.
In this study, we aimed to evaluate the role of P. babylonica extract on serum blood glucose, total cholesterol, triglycerides, and low- and high-density lipoprotein-cholesterol in STZ-induced diabetes in rats.
Materials and methods
Drugs, Reagents, and Instruments:
1,1-diphenyl-2-picrylhydrazyl (DPPH) and streptozotocin (STZ) were purchased from sigma-Aldrich Inc. Standard kits for total cholesterol, high density lipoprotein (HDL), low density lipoprotein (LDL) and triglycerides (TGs) were purchased from Labtest® diagnostica. All other chemicals of analytical grade were purchased from standard commercial suppliers.
Plant Materials
babylonica was collected from Riyadh region, Saudi Arabia. Aerial parts of plant material were air-dried in accordance with the universal standard herbarium procedures. A plant voucher specimen is kept in Department of Pharmaceutical Sciences, College of Clinical Pharmacy, King Faisal University for future research and reference.
Plant Extract Preparation
The air-dried ground of P. babylonica aerial parts (1 kg) was exhaustively extracted three times with 3 days time interval at the room temperature (21-23 °C) using 10 Liters of 70% methanol in water through cold maceration method at the room temperature. The solvent was removed by rotary evaporator 12. The resultant extract was directly freeze-dried to yield the total extract (90 g).
Radical scavenging DPPH assay
The protocol of the DPPH free radical scavenging assay was adapted with some modifications from [17-19]. Briefly, the tested extract was initially dissolved in 100 µl of methanol in a 96-well microplate, The absorbance (Ab) of the test extract was recorded at 515 nm at zero time (t0) as Ab blank. Then, 100 µl of 200 µM DPPH solution was added to every well, and then was kept in the darkness at the room temperature for 30 minutes. Subsequently, the absorbance of the tested extract was measured again as Ab sample. The inhibition percentage (% of inhibition) was measured using the following equation:
% of inhibition = (1 – [Ab sample – Ab blank] / [Ab control – Ab blank]) ×100
Where Ab control is the absorbance of a mixture of all other reactants and DMSO without the tested extract. Half-maximal inhibitory concentration (IC50) was determined as the required concentration of tested sample to cause inhibition of 50 % DPPH free radicals in the method of DPPH radical scavenging.
Experimental Animals
Thirty Wistar rats of both sexes weighing between 150-200 g were obtained from the animal care facility, King Faisal University, Saudi Arabia. Animals were maintained for 1 week to be acclimatized, i.e., ambient room temperatures of (23±2) ºC, at relative humidity of 45-55% for 12 hr time intervals, each of dark and light cycle. The animals were fed with standard rodent pellets and water ad libitum before the start of experiment. All procedures that involve experimental animals were carried out in according to the guidelines on the usage and caring of experimental animals, published by the US National Institutes of Health (NIH) and approved by the Committee of Institutional Research Ethics (with protocol ID: KFU-REC-2021-NOV-EA000166) at King Faisal University, Saudi Arabia.
Induction of diabetes
Experimental animals were exposed to a high fat diet for two weeks and subsequently a single intraperitoneal (IP) dose of 40 mg/kg body weight of STZ to induce diabetes as described in Furman et.al with some modifications [20]. The fresh preparation of STZ was in 0.1 mol/L of cold sodium citrate buffer at a pH of 4.5 for stability enhancement [21, 22]. The confirmation of diabetes development was by measuring the blood glucose level before and 72 hours after the injection of STZ. Rats with a basal blood glucose level more than 200 mg/dL were used in the experimental study [23].
Experimental design
A total of 30 experimental rats, including 6 normal control and 24 STZ-induced diabetic surviving rats, were randomly divided in the following five groups with 6 animals per group:
Group I: Normal healthy control rats.
Group II: Diabetic control rats.
Group III: Diabetic rats received P. babylonica extract with a daily dose of 200 mg/kg body weight.
Group IV: Diabetic rats treated with P. babylonica extract with a daily dose of 400 mg/kg body weight.
Group V: Diabetic rats received Glibenclamide with a daily dose of 0.6 mg/kg body weight [24].
The treatment by the P. babylonica extract was dissolved in 5% carboxy methyl cellulose (CMC) solutions and given orally for two weeks in Group III and Group IV, and Glibenclamide in Group V.
The experimental animals were euthanized at the end of the experiment. The blood glucose level was measured by blood glucometer (One Touch Ultra, LifeScan, Milpitas, CA, USA). The level determination of some parameters of lipid profile in serum that includes total cholesterol, serum triglycerides (TGs), high-density lipoprotein (HDL) cholesterol and low-density lipoprotein (LDL) cholesterol levels were performed by enzymatic colorimetric methods (UV/Vis) using commercial kits (Labtest® diagnostica) according to the manufacturer’s instructions.
Results
DPPH radical scavenging activity
The examination of the radical scavenger activity of plant total extract was performed by using DPPH free radical scavenging assay. It showed marked scavenging activity (IC50:29.4 µg/mL), comparable with the standard trolox (IC50:21.8 µM).
Effect of P. babylonica on glucose level after 1 and 2 weeks of treatment
The potentials of antihyperglycemic effect of P. babylonica extract-treated groups compared to healthy control, diabetic control and standard glibenclamide-treated groups were investigated in STZ-induced diabetic rats at different time intervals (Figure 1). Results revealed that the measured level of blood glucose in diabetic control (Group II) was slightly increasing with time from 225.58 ± 6.97 to 243.12 ± 4.89 mg/dL in comparison with the healthy control group (Group I). Generally, the highest blood glucose level was found in diabetic control (Group II) compared to the groups of P. babylonica extract 200 mg/kg (Group III), P. babylonica extract 400 mg/kg (Group IV), and standard glibenclamide (Group V) at all time intervals after the diabetes induction. Our major observation from the obtained results is that the oral administration of 200-400 mg/kg P. babylonica extract exhibited a significant antihyperglycemic effect. Moreover, experimental animals treated with 200 mg/kg dose of P. babylonica extract (Group III) showed a substantial reduction in the measured blood glucose concentrations between day 1 and day 14, which was 226.45 ± 6.71 to 132.43 ±5.28 mg/dL in comparison with diabetic control (Group II). Similarly, the experimental rats treated with 400 mg/kg dose of P. babylonica extract (Group IV) had a decrease in the measured blood glucose level in the allocated time-period from 230.25 ± 6.34 to 124.52 ± 4.33 mg/dL in comparison with diabetic control (Group II). Therefore, we can attribute that the observed antihyperglycemic effect to be because of the administration of the extract of P. babylonica plant. However, when compared to the rats treated with standard glibenclamide, the animals treated with P. babylonica in both III and IV groups had less antihyperglycemic effect.
 |
Figure 1: Effect of daily oral administration of the extract of P. babylonica on blood glucose in STZ-induced diabetic rats. |
Effect of P. babylonica on dyslipidemia
The lipid profile of all groups was determined on the last day of the experiment (after 14 days of treatment), as shown in Figure 2. Biochemical analysis of the blood serum, from diabetic control (Group II), has shown a substantial increase in levels of total cholesterol, TGs, as well as LDL, but there was a decrease in HDL level compared to the healthy rats (Group I). Generally, our observation is that animals treated with P. babylonica extract (Group III and IV) had a significant reduction in LDL, TGs, and total cholesterol, however, level of HDL was higher, compared to diabetic control (Group II). As shown in the results, it appears to exhibit a dose-dependent effect with P. babylonica extract 400 mg/kg (Group IV) have the highest antihyperlipidemic effect and P. babylonica extract 200 mg/kg (Group III) have the least antihyperlipidemic effect, but it is still significant different when compared to diabetic control (Group II). Moreover, the animals treated with P. babylonica extract at 200 and 400 mg/kg doses (Group III and IV) had less antihyperlipidemic effect in comparison with the rats treated with standard glibenclamide. Therefore, these results indicate that P. babylonica extract might have contributed to lowering total cholesterol, LDL, and TGs levels.
 |
Figure 2: Effect of oral administration of extract of P. babylonica on lipid profile in STZ-induced diabetic rats after 14 days. |
Discussion
Type 2 DM accounts for approximately 90% of all diabetes and considered one of the most prevalent metabolic disease in the world [1]. Moreover, there is an association between the uncontrolled type 2 DM, due to treatment failure, and a high risk of developing dyslipidemia, as well as cardiovascular diseases[3, 5]. This increases the necessity to investigate for an effective antidiabetic from natural products with little side effects. Previously, numerous experiments have been carried out to investigate antidiabetic and hypolipidemic potentials of natural extracts in experimental animal models using rats and mice. It is well known that STZ, a nitrosourea compound produced by Streptomyces achromogenes, can cause DM by inducing DNA strand breakage in pancreatic β-cells [25]. As a result, the level of insulin will decline, leading to augmentation of glucose level in the blood. In current study, P. babylonica extract demonstrated antidiabetic potential by significantly reducing the level of plasma glucose to 132.43 ± 5.28 mg/dL and 124.52 ± 4.33 mg/dL with both doses of 200 mg/kg and 400 mg/kg, respectively, in a dose-dependent manner.
Several reports indicated that DM is highly associated with disturbance in lipid profiles. Indeed, DM considered the main risking factor in hypertriglyceridemia and hypercholesteremia. In addition, we have shown that total cholesterol, LDL and TGs levels were significantly increased in diabetic control (Group II) with the following values: 186.43 ±1.61 mg/dL; 138.9 ± 1.26 mg/dL; and 173.2 ± 1.53 mg/dL, respectively, while the HDL level was reduced. We demonstrated that extract-treated groups remarkably diminished total cholesterol, LDL and TGs levels. Also, we elucidated that extract-treated groups improved HDL level, which was 37.39 ± 1.5 mg/dL with P. babylonica extract 200 mg/kg and 43.07 ± 1.83 mg/dL with P. babylonica extract 400 mg/kg. These results indicate the potential hypolipidemic effects of P. babylonica extract.
Insulin normally activates lipoprotein lipase enzyme and hydrolyses TGs, but in insulin deficiency will hinder these physiological processes causing hypertriglyceridemia. The mechanism of anti-hyperlipidemic effect by P. babylonica extract is proposed to be by inhibiting fatty acid synthesis, However, further research is necessary to fully examine the precise mechanism. It is important to mention that one of this study limitations is that the measurement of the subjects’ body weight were not reported before and after the treatment. From these findings, we can conclude that P. babylonica extracts have significant antidiabetic and hypolipidemic properties. Nevertheless, more confirmatory research is recommended to investigate the plausible involvement of P. babylonica active constituents pertaining these properties.
Conclusion
The present study revealed that the P. babylonica extract could be developed as an antidiabetic and anti-dyslipidemic agent. However, further research is warranted to discover the active constitutes as well as to investigate the possible molecular mechanism.
Acknowledgment
The Author highly acknowledges Dr. Hany Ezzat Khalil for help in the preparation of plant extract and also extends the appreciation to the Department of Pharmaceutical Sciences, College of Clinical Pharmacy, King Faisal University for the encouragement and moral support throughout the study.
Conflict of Interest
The author reports no conflicts of interest.
Funding Sources
This study was partially supported by the department of pharmaceutical sciences, King Faisal University.
References
- International Diabetes Federation, IDF Diabetes Atlas. 10 ed, ed. I.D. Federation. 2021, Brussels, Belgium.
- American Diabetes Association, Diagnosis and classification of diabetes mellitus. Diabetes care, 2013. 36 Suppl 1(Suppl 1): p. S67-S74.
CrossRef - World Health Organization, Classification of diabetes mellitus. 2019, Geneva: World Health Organization.
- Cuschieri, S., Type 2 Diabetes: An Unresolved Disease, in Obesity and Diabetes: Scientific Advances and Best Practice, J. Faintuch and S. Faintuch, Editors. 2020, Springer International Publishing: Cham. p. 567-578.
CrossRef - Martín-Timón, I., et al., Type 2 diabetes and cardiovascular disease: have all risk factors the same strength? World journal of diabetes, 2014. 5(4): p. 444.
CrossRef - Chaudhury, A., et al., Clinical Review of Antidiabetic Drugs: Implications for Type 2 Diabetes Mellitus Management. Front Endocrinol (Lausanne), 2017. 8: p. 6.
CrossRef - Chandramohan, G., S. Ignacimuthu, and K.V. Pugalendi, A novel compound from Casearia esculenta (Roxb.) root and its effect on carbohydrate metabolism in streptozotocin-diabetic rats. Eur J Pharmacol, 2008. 590(1-3): p. 437-43.
CrossRef - Kim, S.H., S.H. Hyun, and S.Y. Choung, Anti-diabetic effect of cinnamon extract on blood glucose in db/db mice. J Ethnopharmacol, 2006. 104(1-2): p. 119-23.
CrossRef - Kesari, A.N., et al., Studies on the glycemic and lipidemic effect of Murraya koenigii in experimental animals. Journal of Ethnopharmacology, 2007. 112(2): p. 305-311.
CrossRef - Sidhu, M. and T. Sharma, A Database of Antidiabetic Plant Species of Family Asteraceae, Euphorbiaceae, Fabaceae, Lamiaceae and Moraceae. International Journal of Herbal Medicine, 2013. 1: p. 139.
CrossRef - Trojan-Rodrigues, M., et al., Plants used as antidiabetics in popular medicine in Rio Grande do Sul, southern Brazil. J Ethnopharmacol, 2012. 139(1): p. 155-63.
CrossRef - Khalil, H.E., QUALITATIVE ASSESSMENT OF CHEMICAL CONSTITUENTS OF PICRIS BABYLONICA. World journal of pharmaceutical research, 2017: p. 719-725.
CrossRef - Michalska, K., E. Szneler, and W. Kisiel, Sesquiterpenoids of Picris koreana and their chemotaxonomic significance. Biochemical Systematics and Ecology – BIOCHEM SYST ECOL, 2007. 35: p. 459-461.
CrossRef - Mirjana, M., et al., Antioxidant activities of the constituents of Picris echoides. Journal of the Serbian Chemical Society, 2002. 67.
CrossRef - Milovanovic, M. and K. Picuric-Jovanovic, Terpenoids from Picris echoides. Fitoterapia, 2005. 76(5): p. 490-2.
CrossRef - Jia, Y. and L. Chen, Antidiabetic Activity of Picris japonica Thunb Aqueous Extract in Diabetic KK-A(y) Mice. Evid Based Complement Alternat Med, 2018. 2018: p. 1298030.
CrossRef - Muniyappan, N. and N. Nagarajan, Green synthesis of silver nanoparticles with Dalbergia spinosa leaves and their applications in biological and catalytic activities. Process biochemistry, 2014. 49(6): p. 1054-1061.
CrossRef - Khalil, H. and A. Al-Ahmed, Phytochemical Analysis and Free Radical Scavenging Activity of Carthamus oxyacantha Growing in Saudi Arabia: A Comparative Study. 2017. 45: p. 51-55.
- Yong, Y.K., et al., Clinacanthus nutans extracts are antioxidant with antiproliferative effect on cultured human cancer cell lines. Evidence-Based Complementary and Alternative Medicine, 2013. 2013.
CrossRef - Furman, B.L., Streptozotocin-Induced Diabetic Models in Mice and Rats. Curr Protoc, 2021. 1(4): p. e78.
CrossRef - Akbarzadeh, A., et al., Induction of diabetes by streptozotocin in rats. Indian Journal of Clinical Biochemistry, 2007. 22(2): p. 60-64.
CrossRef - Srinivasan, K. and P. Ramarao, Animal model in type 2 diabetes research: An overview. Indian Journal of Medical Research, 2007. 125(3): p. 451.
- Qinna, N.A. and A.A. Badwan, Impact of streptozotocin on altering normal glucose homeostasis during insulin testing in diabetic rats compared to normoglycemic rats. Drug design, development and therapy, 2015. 9: p. 2515.
CrossRef - Balamurugan, R. and S. Ignacimuthu, Antidiabetic and Hypolipidemic effect of methanol extract of Lippia nodiflora L. in streptozotocin induced diabetic rats. Asian Pacific Journal of Tropical Biomedicine, 2011. 1(1, Supplement): p. S30-S36.
CrossRef - Thangaraj, P., Evaluation of Anti-diabetic Property on Streptozotocin-Induced Diabetic Rats, in Pharmacological Assays of Plant-Based Natural Products. 2016, Springer International Publishing: Cham. p. 145-149.
CrossRef
