Abstract
BACKGROUND:
The incidence of diabetes and its complications has risen since the last decades, and it has been predicted that the prevalence of diabetes will be double in the coming year. Chronic hyperglycemia marked as elevated advanced glycation end products plays an important role in complications of diabetes. Basil leaves (Ocimum basilicum) originating from India and commonly found in Southeast Asia, is widely used empirically to decrease the blood sugar in diabetic patients.
AIM:
The purpose of the study was to investigate the effect of extract ethanol of Basil leaves in diabetic rats.
METHODS:
This is an experimental study; Streptozotocin was used to induce type 2 diabetic rats. Thirty Wistar male rats were divided into six groups (normal group, group treated with metformin 45 mg/kg body weight, group treated with Basil leaves 100 mg/kg body weight, 200 mg/kg body weight, 400 mg/kg body weight, control diabetic group). The blood glucose level was measured before induction and after treatment at week 1 and 4. Advanced glycation end products were measured in all groups at week 4.
RESULTS:
Extract ethanol of basil leaves 100, 200, 400 mg/body weight all showed significant lowering blood glucose level (p < 0.001) compared to diabetic group without treatment, but did not show superior to metformin group (p = 0.17), advanced glycation end products did not show statistical significance between group with normal rats and group of diabetic rats treated with basil leaves 100 mg/kg, 200 mg/kg, 400 mg/kg (p = 0.12, p = 0.33, p = 0.26).
CONCLUSION:
Extract ethanol of basil leaves showed good results in lowering blood glucose and advanced glycation end products in diabetic rats.
Keywords: Basil leaves, Blood glucose, Advanced glycation end products, Diabetic Rats
Introduction
Diabetes Mellitus (DM) is a chronic metabolic disease manifested with hyperglycemia caused by insulin deficiency or resistance. Prolonged elevation of the blood glucose level will cause macro and microvascular complications that resulted in damage of various organs of the body. More than eighty per cent of these conditions were found in low- or middle-income countries [1]. It is estimated that in 2030, in every ten adults one person will get diabetes mellitus (DM) [2]. Chronic hyperglycemia may induce glycation of various plasma proteins, and collagen [3] thus advanced glycation end-products (AGEs) are largely produced and accumulated in various tissues and blood circulation [4]. This result of protein formation through non-enzymatic processes caused by high blood sugar levels are thought to alter the hemostatic system, activated coagulation system and generation of oxygen free radicals. Advanced glycation is one of the major causes of diabetic complications [5], [6]. Control diets, exercise and medicine, have been the mainstay treatments of diabetes [7], [8], [9]. Most of the people in Indonesia still used traditional medicine empirically to treat diabetes, like Poguntano leaves [10] as Indonesia is a country of plenty of medicinal plants. Basil leaves (Ocimum basilicum) from family Lamiaceae (mints) known as the king of herbs or “royal herb” originating from India and commonly found in Southeast Asia, is widely used as a culinary ingredient [11], [12].
Empirically, it was used widely to decrease blood sugar in diabetic patients. Basil leaves were reported to have antihyperglycemic and liver protective effects as it stimulates the release of insulin from the pancreas, inhibit the glucose production in the liver and increased glycogen synthesis [13], [14].
This study aimed to study the effect of basil leaves on blood glucose level and AGE in diabetic rats.
Methods
This is an experimental study using Streptozotocin (STZ) in a dose of 40 mg/body weight injected intraperitoneal to induce type 2 diabetic rats. Thirty Wistar male rats weighed 150-200 grams were divided into six groups (normal group, group treated with metformin in a dose of 45 mg/kg body weight, group treated with basil leaves 100 mg/kg body weight, 200 mg/kg body weight, 400 mg/kg body weight, control diabetic group) for four weeks. Blood glucose levels were measured before starting the experiment; after the induction, blood glucose levels were measured in all groups at week 1, 4 and AGE level were measured in all groups at week 4.
This study will be analysed using SPSS V20 with One Way ANOVA.
Preparation of basil extract
Extraction was carried out by maceration using 96% ethanol as solvent standardisation of extracts including the determination of water content, levels of water-soluble and ethanol extracts, total ash content, insoluble acid and chromatogram profiles [15].
Laboratory Assay
Blood Glucose was measured using spectrophotometry, and AGE using sandwich Elisa with a double antibody.
Results
One way ANOVA was used to analyse the blood glucose levels at week 4 within groups, and it showed statistically significant (p = 0.0). Post hoc was used to analysed further and found out that the blood glucose level was statistically significant differences within the diabetic and treatment groups (p = 0.00) but did not show the statistical difference (p = 1) within treatment groups (metformin and basil groups).
Table 1.
Group A | Group B | Group C | Group D | Group E | Group F | |
---|---|---|---|---|---|---|
Blood Glucose level (mg/dl) | 78 ± 2 | 79.3 ± 1.5 | 78 ± 2 | 80 ± 2 | 79.3 ± 2 | 81. 3 ±1.5 |
Note: Group A: normal control, Group B: Diabetic Control, Group C: a group treated with metformin, Group D: group treated with basil 100 mg, Group E: a group treated with basil 200 mg, Group F: a group treated with basil 400 mg.
Advanced glycation end products showed statistical significant within diabetic control group and group treated either with metformin or basil leaves (p = 0.01), but did not showed statistical significance between group with normal rats and group of diabetic rats treated with basil leaves extracts 100mg/kg, 200 mg/kg, 400 mg/kg (p = 0.12, p = 0.33, p = 0.26).
Table 2.
Group A | Group B | Group C | Group D | Group E | Group F | |
---|---|---|---|---|---|---|
T1 (mg/dl) | 80 ± 1 | 501 ± 6.1 | 553 ± 5 | 473 ± 2.5 | 531 ± 4.1 | 525 ± 7.8 |
T4 (mg/dl) | 80 ± 1.5 | 495 ± 4.6 | 92 ± 4 | 131 ± 6.4 | 123 ± 3 | 107 ± 3 |
AGE (ng/ml) | 8.5 ± 1.3 | 5.6 ± 1.4 | 17 ± 1.0 | 4.6 ± 1.8 | 5.5 ± 1.8 | 5.2 ± 1.3 |
Notes: T1: blood glucose level after 1 week STZ injection; T4: blood glucose level week 4.
Discussion
Basil leaves extract showed statistical significant lowering blood glucose between the diabetic control group and the group treated with metformin or basil leaves either in 100, 200, 400 mg/kg body weights (p = 0.0), but did not show a statistical difference within the treatment group (p = 1). This means that either metformin or basil leaves can be used for lowering blood glucose, and one is not inferior to the other; this has been reported by Eddouks M and Kumar S that basil leaves have the ability as anti-diabetic agents and improve the insulin sensitivity [16], [17].
Advanced glycation end products showed a significant difference between the diabetic control group and group treated either with metformin or basil leaves; these results showed that the Basil leaves could also be used to treat the chronic hyperglycemic effect of diabetes, one of the most important issues in diabetes.
In the basil treatment group, we do not find any inferior effect in lowering blood glucose level in the three groups (100 mg, 200 mg, 400 mg/kg body weights).
In conclusion, extract ethanol of basil leaves showed good results in lowering blood glucose and advanced glycation end products in diabetic rats.
Footnotes
Funding: This study was supported by the Ministry of Research and Technology and the Higher Education Republic of Indonesia, under research grant DRPM
Competing Interests: The authors have declared that no competing interests exist
References
- 1.American Diabetes Association. New York: Diagnosis and classification of diabetes mellitus; 2009. [Google Scholar]
- 2.World Health Organization. Diagnosis and classification of diabetes mellitus and its compilation. Geneva: Department of Noncommunicable Disease Surveillance; 2008. [Google Scholar]
- 3.Varun Parkash Singh, Anjana Bali, Nirmal Singh, Amteshwar Singh Jaggi. Advanced Glycation End Products and Diabetic Complications. Korean J Physiol Pharmacol. 2014;18:1–14. doi: 10.4196/kjpp.2014.18.1.1. https://doi.org/10.4196/kjpp.2014.18.1.1 PMid:24634591 PMCid:PMC3951818. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4.Sang youl Rhee, Young Seoul Kim. The Role of Advanced Glycation End Products in Diabetic Vascular Complications. Diabetes Metab J. 2018;42(3):188–195. doi: 10.4093/dmj.2017.0105. https://doi.org/10.4093/dmj.2017.0105 PMid:29885110 PMCid:PMC6015964. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5.Negre-Salvayre A, Salvayre R, Augé N, Pamplona R, Portero-Otín M. Hyperglycemia and glycation in diabetic complications. Antioxid Redox Signal. 2009;11(12):3071–109. doi: 10.1089/ars.2009.2484. https://doi.org/10.1089/ars.2009.2484 PMid:19489690. [DOI] [PubMed] [Google Scholar]
- 6.Fraser DA, Hansen KF. Making sense of advanced glycation end products and their relevance to diabetic complications. Inter Diabetes Monitor. 2005;17:1–7. [Google Scholar]
- 7.Joshua A, Beckman I, Francesco P, Francesco C, Mark A, Crager Diabetes and vascular disease:pathophysiology clinical consequences, and medical therapy:part II. European Heart Journal. 2013;34(31):2436–43. doi: 10.1093/eurheartj/eht149. https://doi.org/10.1093/eurheartj/eht149 PMid:23641007 PMCid:PMC3743069. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 8.Ley SH, Hamdy O, Mohan V, Hu FB. Prevention and management of type 2 diabetes:dietary components and nutritional strategies. Lancet. 2014;383(9933):1999–2007. doi: 10.1016/S0140-6736(14)60613-9. https://doi.org/10.1016/S0140-6736(14)60613-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9.Nishida C, Uauy R, Kumanyika S, Shetty P. The joint WHO/FAO expert consultation on diet, nutrition and the prevention of chronic diseases:process, product and policy implications. Public Health Nutr. 2004;7(1A):245–50. doi: 10.1079/phn2003592. [DOI] [PubMed] [Google Scholar]
- 10.Widjaja SS, Rusdiana Extract Ethanol of Poguntano in Alloxan Induced Diabetic Rats. Bangladesh Journal of Medical science. 2018;17(2):251–54. https://doi.org/10.3329/bjms.v17i2.35879. [Google Scholar]
- 11.EL-Beshbishy HA, Bahashwan SA. Hypoglycemic effect of basil (Ocimum basilicum) aqueous extracts mediated through inhibition of α-glucosidase and α-amylase activities. Toxicol Ind Health. 2012;28(1):42–50. doi: 10.1177/0748233711403193. https://doi.org/10.1177/0748233711403193 PMid:21636683. [DOI] [PubMed] [Google Scholar]
- 12.Yacout G, Elguiindy N, Azab E. Hepatoprotective effect of basil (Ocimum basilicum L.) On CCl4- induced liver fibrosis in rats. Afr J Biotech. 2012;2(90):15702–11. https://doi.org/10.5897/AJB12.2048. [Google Scholar]
- 13.Xu Z, Wang X, Zhou M, Ma L, Deng Y, Zhang H, et al. The antidiabetic activity of total lignin from Fructus Arctii against alloxan-induced diabetes in mice and rats. Phytother Res. 2008;22(1):97–101. doi: 10.1002/ptr.2273. https://doi.org/10.1002/ptr.2273 PMid:17724763. [DOI] [PubMed] [Google Scholar]
- 14.Krishnasamy G, Muthusamy K, Chellappan DR, Subbiah N. Antidiabetic, antihyperlipidaemic, antioxidant activity of Syzygium densiflorum fruits in streptozotocin and nicotinamide-induced diabetic rats. Pharm Biol. 2016;54(9):1716–26. doi: 10.3109/13880209.2015.1125932. https://doi.org/10.3109/13880209.2015.1125932 PMid:26704340. [DOI] [PubMed] [Google Scholar]
- 15.BPOM RI. Jakarta: BPOM RI; 2005. Peraturan Kepala Badan Pengawas Obat dan Makanan Republik Indonesia Tentang Kriteria dan Tata Laksana Pendaftaran Obat Tradisional, Obat herbal Terstandar dan Fitofarmaka. [Google Scholar]
- 16.Eddouks M, Bidi A, El Bouhali B, Hajji L, Zeggwagh NA. Antidiabetic plants improving insulin sensitivity. J Pharm Pharmacol. 2014;66(9):1197–214. doi: 10.1111/jphp.12243. https://doi.org/10.1111/jphp.12243 PMid:24730446. [DOI] [PubMed] [Google Scholar]
- 17.Kumar S, Naewal S, Kumar V, Prakash O. α-glucosidase inhibitors from plants:A natural approach to treat diabetes. Pharmacogn Rev. 2011;5(9):19–29. doi: 10.4103/0973-7847.79096. https://doi.org/10.4103/0973-7847.79096 PMid:22096315 PMCid:PMC3210010. [DOI] [PMC free article] [PubMed] [Google Scholar]