|Year : 2022 | Volume
| Issue : 1 | Page : 24-28
Ameliorative activities and safety of Moringa Oleifera oil and Nigella Sativa oil on diet-induced hyperlipidemic male wistar rats
Godwin Atufe1, Oghenetega ThankGod Oweh2, Ohunene Makoju Avidime3, Ejiro Prosper Awhin4, Quadri Olaide Nurudeen5
1 Department of Biochemistry, University of Benin, Benin City, Nigeria
2 Department of Medical Biochemistry, College of Medicine, Kaduna State University, Kaduna State, Nigeria
3 Department of Physiology, College of Medicine, Kaduna State University, Kaduna State, Nigeria
4 Department of Medical Biochemistry, Delta State University, Abraka, Nigeria
5 Department of Biological Sciences (Biochemistry Unit), Al-Hikmah University, Kwara State, Ilorin, Nigeria
|Date of Submission||28-Feb-2022|
|Date of Decision||16-Apr-2022|
|Date of Acceptance||19-Apr-2022|
|Date of Web Publication||01-Jul-2022|
Dr. Oghenetega ThankGod Oweh
Department of Medical Biochemistry, College of Medicine, Kaduna State University KASU
Source of Support: None, Conflict of Interest: None
Background: Numerous medicinal plants have been explored as therapy for hyperlipidemia which could be induced by a high-fat diet (HFD). Aims and Objectives: The ameliorative effects of Moringa oleifera oil and Nigella sativa oil and their safety on diet-induced hyperlipidemic Wistar rats were examined. Materials and Methods: Thirty Wistar rats were distributed into six groups of five each. Group 1 was the control group while the other groups were fed with HFD. Groups 3–6 were treated using 1 ml/kg BW of M. oleifera oil, 0.5 ml/kg BW M. oleifera oil, 1 ml/kg BW N. sativa oil, and 0.5 ml/kg BW N. sativa oil, respectively, for 10 days. The plasma total and tissue cholesterol, triglyceride, and high-density lipoprotein (HDL)-cholesterol concentrations as well as alkaline phosphatase (ALP), alanine aminotransferases (ALT), and aspartate aminotransferase (AST) activities were analyzed. Results: M. oleifera oil and N. sativa oil (1 ml/kg BW and 0.5 ml/kg BW) showed significant reduction at (P < 0.05) in total plasma cholesterol and lipid levels compared to the control group but had no significant effects on the activities of AST, ALT, and ALP. Conclusion: The study proved that M. oleifera and N. sativa oil ameliorate diet-induced hyperlipidemia in Wistar rats by reducing plasma cholesterol, triglyceride, and increasing HDL levels and show no adverse effects on the activities of the liver enzymes.
Keywords: Hyperlipidemia, liver enzymes, plasma cholesterol
|How to cite this article:|
Atufe G, Oweh OT, Avidime OM, Awhin EP, Nurudeen QO. Ameliorative activities and safety of Moringa Oleifera oil and Nigella Sativa oil on diet-induced hyperlipidemic male wistar rats. Niger J Exp Clin Biosci 2022;10:24-8
|How to cite this URL:|
Atufe G, Oweh OT, Avidime OM, Awhin EP, Nurudeen QO. Ameliorative activities and safety of Moringa Oleifera oil and Nigella Sativa oil on diet-induced hyperlipidemic male wistar rats. Niger J Exp Clin Biosci [serial online] 2022 [cited 2022 Sep 28];10:24-8. Available from: https://www.njecbonline.org/text.asp?2022/10/1/24/349558
| Introduction|| |
Hyperlipidemia, which is a disorder that is characterized by the presence of high levels of lipids in the blood, is a major threat that influences cardiovascular disease and is a severe health concern to the world. Numerous clinical studies have explained its role in atherosclerosis pathogenesis. Several attempts are made at exploring the potential of medicinal plants toward the management of hyperlipidemia. Supplements from plants have been reported to be very effective in cardiovascular health and serve to reduce atherosclerosis (caused by high lipid in the blood).
Moringa oleifera and Nigella sativa are two different plants with numerous medicinal activities attached to their various parts such as leaves, roots, stems, and even the oil obtained from these plants. M. oleifera possesses numerous dietary and therapeutic values ascribed to the roots, bark, leaves, flowers, fruits, and seeds. Research has shown that most of the parts of the plant hold hepatoprotective, antimicrobial, and antidiabetic activity. Furthermore, the hypocholesterolemic activity of M. oleifera crude extract was reported. Furthermore, N. sativa Linn. (N. sativa) well known as black seed or black cumin belongs to the botanical family of Ranunculaceae. The seeds of the plant serve as a natural remedy used in the treatment of several diseases such as asthma, hypertension, diabetes, hypercholesterolemia, inflammation, arthritis, tumor, and gastrointestinal disturbances.,, They are broadly used as seasoning and fragrant added to beverages, salads, and bread.
N. sativa has been reported to produce several favorable actions such as hypoglycemic, hypocholesterolemic, and antioxidant effects. N. sativa oils and its active constituent thymoquinone have been reported to have a useful effect in hyperglycemia and hyperlipidemia. Pourghassem-Gargari et al. reported the ameliorative activities of N. sativa seed on diet-induced hyperlipidemic rabbits. Furthermore, Ajayi et al. reported the lipid-altering potentials of M. oleifera seed extract in Wistar rats. Similarly, Souravh et al. reported the hypolipidemic activity of M. oleifera leaves against high-fat diet (HFD)-induced obesity in rats. Literature is replete with the role of the seeds and leaves of both M. oleifera and N. sativa in reducing high blood lipids. However, there is a paucity of information on the ameliorative activities of the oil of M. oleifera and N. sativa that this study aims to address.
| Materials and Methods|| |
M. oleifera and N. sativa Oil: the M. oleifera and N. sativa oil were obtained from Asa organic oil Factory, Benin City, Edo State, Nigeria.
Thirty male Wistar rats with weighing 80–120 g were housed in a standard cage with a room temperature of 25°C and were fed with rat chow and water ad libitum during the 2-week acclimatization.
They were grouped into six groups of five rats each.
- Group 1: Animals that feed on normal diet (control)
- Group 2: HFD only
- Group 3: HFD + 1 ml/kg Bw M. oleifera oil
- Group 4: HFD + 0.5 ml/kg Bw M. oleifera oil
- Group 5: HFD + 1 ml/kg Bw N. sativa oil
- Group 6: HFD + 0.5 ml/kg Bw N. Sativa oil.
All chemicals and reagents used were of analytical grade. Total cholesterol, triglyceride, low-density lipoprotein (LDL), and high-density lipoprotein (HDL)-cholesterol assay kits were products of Randox, United Kingdom.
Induction of hyperlipidemia
The rats in Group 1 were fed with a controlled diet, while the rats in Groups 2–6 were fed with HFD as described by Matos et al. The ingredient of the diet and their composition is shown in [Table 1].
The experiment lasted for 10 days, after which the animals were fasted overnight and anesthetized in a chloroform chamber.
The study was conducted in agreement with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health and all efforts were made to minimize animal suffering and the number of animals used. The protocol was approved by the Ethics Committee of University of Benin, Nigeria.
Collection of blood sample
The blood samples were collected by retro-orbital sinus puncture, under mild chloroform anesthesia in lithium heparin bottles and centrifuged for 10 min at 3000 rpm. The supernatants were stored frozen at –20°C until required for biochemical assays.
Tissue collection/preparation of tissue homogenates
Liver and heart were collected and washed in ice-cold physiological saline repetitively and weighed accurately. A portion of each of the tissue was chopped into very small pieces and homogenized in ice-cold physiological saline (1 g tissue: 5 ml saline – 20% homogenates) using a precooled mortar and pestle. The homogenates were centrifuged at 3000 rpm for 10 min and the supernatant stored frozen at –20°C until required for biochemical assays.
Determination of total cholesterol, HDL-cholesterol, and LDL-cholesterol concentrations
Total cholesterol, HDL-cholesterol (HDL-C), and LDL-cholesterol in the homogenate supernatants were determined using the method described by Allain et al., Roeschlaw et al. National Cholesterol Education Program (NCEP).,,
Determination of triglycerides concentrations
Triglycerides concentrations in the tissue homogenates supernatant were determined following the method described by the NCEP. The method defined in the assay kit leaflets comprises routine enzymatic methods demonstrated by that defined by Stein and Myers.
Estimation of liver enzymes
Alkaline phosphatase (ALP), alanine aminotransferases (ALT), and aspartate aminotransferase (AST) were estimated using the method described by Teitz.
| Results|| |
The results of the study showing the effects of M. oleifera and N. sativa oil on lipid profile parameters as well as its safety regarding liver enzymes are presented in [Figure 1],[Figure 2],[Figure 3],[Figure 4].
|Figure 1: Effects of Moringa oleifera oil and Nigella sativa oil on serum lipid profile of diet-induced hyperlipidemic Wistar rats|
Click here to view
|Figure 2: Effects of Moringa oleifera oil and Nigella sativa oil on lipid profile of liver homogenate on diet-induced hyperlipidemic Wistar rats|
Click here to view
|Figure 3: Effects of Moringa oleifera oil and Nigella sativa oil on lipid profile of heart homogenate on diet-induced hyperlipidemic Wistar rats|
Click here to view
|Figure 4: Effects of Moringa oleifera oil and Nigella sativa oil on serum liver function parameters of diet-induced hyperlipidemic Wistar rats|
Click here to view
The HFD increased significantly (P < 0.05) in total cholesterol and triglyceride in the rats compared to the control group. Furthermore, the groups treated with 1 ml/kg BW and 0.5 ml/kg BW of M. oleifera, as well as 1 ml/kg BW N. sativa oil significantly (P < 0.05) reversed the effect of the HFD on serum lipid profile [Figure 1] while the group treated with 0.5 ml/kg BW of N. sativa oil increased significantly (P < 0.05) serum cholesterol compared to the HFD group and control group. However, the groups treated with 1 ml/kg BW, 0.5 ml/kg BW M. oleifera, and 0.5 ml/kg BW N. sativa oil reversed the HFD-induced increase in the lipid profile parameters monitored [Figure 2].
Effects of Moringa oleifera and Nigella sativa oil on liver function enzymes
HFDs increased plasma ALP activity and lowered ALP activity in liver homogenate. However, it had no effect on both AST and ALT. Furthermore, 0.5 and 1 ml/kg BW of M. oleifera oil and 0.5 ml/kg BW of N. sativa oil had no significant effect on the plasma liver enzymes monitored while the group treated with 1 ml/kg BW of N. sativa oil showed a significant increase in the plasma liver enzymes monitored.
| Discussion|| |
This study was conducted to evaluate the hypolipidemic effect of M. oleifera oil and N. sativa oil on HFD fed Wistar rats. The results obtained from this study show that feeding animals with HFD resulted in increased total cholesterol and triglyceride concentrations in serum and liver homogenate. Furthermore, feeding the animals with HFD increased triglycerides and lowered total cholesterol in heart homogenate in this study. Fatty-rich foods have been frequently linked with raised serum cholesterol concentration hypercholesterolemia in animal models., Studies have shown that a high serum concentration of total cholesterol, LDL-cholesterol, and triglyceride coupled with a low HDL-C increase the likelihood of coronary heart disease. The HFD-induced hyperlipidemia is basically as a result of the increase in mobilization of free fatty acids from the peripheral store. The result of this study shows a significant decrease in hyperlipidemia induced by fatty-rich diets. It has been reported severally that serum cholesterol, triglyceride, and HDL are elevated during the consumption of HFDs. M. oleifera leaves extract has been reported to lower serum cholesterol, triglyceride, and LDL cholesterol in HFD-induced hyperlipidemia. M. oleifera oil is a vegetable oil with high contents of monounsaturated fatty acid. It is obtained from M. oleifera seed with high crude fat content of about 20%–40%. It contains a large proportion of oleic acid. Research has shown that M. oleifera has nutritional benefits, various lipids lowering, and antioxidant effects. In this study, serum, liver, and homogenate total cholesterol, triglyceride, and HDL-C were measured in HFD fed rats treated with different doses of M. oleifera oil and N. sativa oil. These parameters are indicators of cardiovascular-related disorders, arteriosclerosis, and other metabolic disorders. The decrease in total cholesterol and triglyceride after the administration of M. oleifera oil and N. sativa oil in this study agrees with the report of Souravh et al. and Farouq et al., who reported a decrease in serum cholesterol, triglyceride, and LDL after treatment with different doses of M. oleifera leave extract. Therefore, the observed decrease in total cholesterol and triglyceride in this study is a pointer to the cardiovascular protective properties of M. oleifera oil and N. sativa oil. An increased level of triglyceride is a major risk factor since it stimulates the deposition of lipid and clotting mechanisms.
Furthermore, the increase in serum HDL-C concentration in rats administered M. oleifera oil and N. sativa oil indicates the protective function of both oil against cardiovascular diseases because HDL-C is produced in the liver and intestine cells and promotes the efflux of cholesterol from the peripheral tissues to the liver, thereby reducing the uptake of cholesterol by macrophages and providing a protective effect against atherosclerosis. Therefore, one major way to reduce the risk of cardiovascular disease is to increase HDL-C levels as observed in this study. The increase in serum HDL-C in this study agrees with the report of Ajayi et al. that reported an increase in serum HDL-C upon the administration of M. oleifera seed and the report of El-Dakhakhny et al., who also reported an increase in serum HDL-C after administering N. sativa oil to male rabbits
Numerous studies have revealed that HFD elevates liver enzymes and causes fatty disease and liver injury. The report from this study, however, revealed that the various doses of M. oleifera oil and N. sativa oil caused no significant alteration in the activities of ALP, ALT, and AST in the serum. This does not correspond with the report of Das et al., who reported a significant decrease in the activities of liver enzymes after administration of M. oleifera oil and Rashidmayvan et al., who also reported a decrease in the activities of liver enzymes after administration of N. sativa oil to patients with nonalcoholic liver disease
| Conclusion|| |
Administration of M. oleifera oil and N. sativa oil on HFD-induced hyperlipidemic rats lowers total cholesterol and triglyceride concentrations and increases HDL-C concentrations after 10 days. Therefore, M. oleifera oil and N. sativa oil could be of immense benefit in ameliorating hyperlipidemia and preventing lipid-related disorders without posing any threat to liver enzymes. Hence, M. oleifera oil and N. sativa oil have strong potential to be explored as a therapeutic agent in lipid-related disorder.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Jaffar AR, Babb J, Movahed A. Optimal management of hyperlipidemia in primary prevention of cardiovascular disease. Int J Cardiol 2004;97:355-66.
Mahmood ZA, Sualeh M, Mahmood SB, Karim MA. Herbal treatment for cardiovascular disease the evidence based therapy. Pak J Pharm Sci 2010;23:119-24.
Kumar SP, Mishra D, Ghosh G, Panda CS. Medicinal uses and pharmacological properties of Moringa oleifera
. Int J Phytomed 2010;2:210-6.
Babu R, Chaudhuri M. Home water treatment by direct filtration with natural coagulant. J Water Health 2005;3:27-30.
Ghasi S, Nwobodo E, Ofili JO. Hypocholesterolemic effects of crude extract of leaf of Moringa oleifera
Lam in high-fat diet fed wistar rats. J Ethnopharmacol 2000;69:21-5.
Ali BH, Blunden G. Pharmacological and toxicological properties of Nigella sativa
. Phytother Res 2003;17:299-305.
El-Din K, El-Tahir H, Bakeet DM. The black seed (Nigella sativa
Linnaeus) – A mine for multi cures: A plea for urgent clinical evaluation of its volatile oil. JTU Med Sci 2006;1:1-19.
Ramadan MF. Nutritional value, functional properties and nutraceutical applications of black cumin (Nigella sativa
L.): An overview. Int J Food Sci Tech 2007;42:1208-18.
Pourghassem-Gargari B, Ebrahimzadeh-Attary V, Rafraf M, Gorbani A. Effect of dietary supplementation with Nigella sativa
L. on serum lipid profile, lipid peroxidation and antioxidant defense system in hyperlipidemic rabbits. J Med Plant Res 2009;3:815-21.
Ajayi TJ, Jones M, Odumuwagun OJ, Joseph AO. Lipid altering potential of Moringa oleifera
lam seed extract and isolated constituents in wistar rats. Afr J Biomed Res 2020;23:77-85.
Bais S, Singh GS, Sharma R. Antiobesity and hypolipidemic activity of Moringa oleifera leaves against high fat diet-induced obesity in rats. J Adv Biol. 2014, Article ID 162914. doi: 10.1155/2014/162914.
Matos SL, de Paula H, Pedrosa ML, dos Santos RC, de Oliveira EL, Chianca Júnior DE, et al.
Dietary models for inducing hypercholesterolemia in rats. Braz Arch Biol Technol 2005;48:203-9.
Allain CC, Poon LS, Chan SG, Richmond W, Fu PC. Enzymatic determination of total serum cholesterol. Clin Chem 1974;20:470-5.
Roeschlaw P, Bernt E, Gruber W. Enzymatic determination of total cholesterol in serum. J Clin Chem Clin Biochem 1974;12:227.
National Cholesterol Education Programme (NCEP). HDL, LDL and TG assay, in PMC; 2001.
Stein EA, Myers GL. National cholesterol education program recommendations for triglycerides measurements: Executive summary. The National Cholesterol Education Program Working Group on Lipoprotein Measurement. Clin Chem 1995;41:1421-6.
Tietz NW. Clinical Guide to Laboratory Tests (ELISA). 3rd
ed. Philadelphia: W.B. Saunders, Co.; 1995. p. 22-3.
Cherng JY, Shih MF. Preventing dyslipidemia by Chlorella pyrenoidosa in rats and hamsters after chronic high fat diet treatment. Life Sci 2005;76:3001-13.
Martinello F, Soares SM, Franco JJ, Santos AC, Sugohara A, Garcia SB, et al.
Hypolipemic and antioxidant activities from Tamarindus indica
L. pulp fruit extract in hypercholesterolemic hamsters. Food Chem Toxicol 2006;44:810-8.
Parab RS, Mengi SA. Hypolipidemic activity of Acorus calamus
L. in rats. Fitoterapia 2002;73:451-5.
Ahmed I, Lakhani MS, Gillet M, John A, Raza H. Hypotriglyceridemic and hypocholesterolemic effects of anti-diabetic Momordica charantia (karela) fruit extract in streptozotocin-induced diabetic rats Diabetes Res Clin Pract 2001;51:155-61.
Farouq F, Meenu R, Avinash T, Abdularif K, Shaila F. Medicinal properties of Moringa oleifera
: An overview of promising healer. J Med Plant Res 2012;6:4368-74.
Harnafi H, Aziz M, Amrani S. Sweet basil (Ocimium basilicum
L.) Improves lipid metabolism in hypercholesterolemic rats. E-SPEN Eur J Clin Nutr Metab 2009;4:e181-6.
Nicholls SJ. Relationship between LDL, HDL, blood pressure and atheroma progression in the coronaries. Curr Opin Lipidol 2009;20:491-6.
El-Dakhakhny M, Mady NI, Halim MA. N. sativa
L oil protects against induced hepatotocity and improves serum lipid profile in rats. ARZNEMTTEL FORSCH 2000;50;832-6.
Das N, Sikder K, Ghosh S, Fromenty B, Dey S. Moringa oleifera
Lam. leaf extract prevents early liver injury and restores antioxidant status in mice fed with high-fat diet. Indian J Exp Biol 2012;50:404-12.
Rashidmayvan M, Mohammadshahi M, Seyedian SS, Haghighizadeh MH. The effect of Nigella sativa
oil on serum levels of inflammatory markers, liver enzymes, lipid profile, insulin and fasting blood sugar in patients with non-alcoholic fatty liver. J Diabetes Metab Disord 2019;18:453-9.
[Figure 1], [Figure 2], [Figure 3], [Figure 4]