International Journal of Biological and Physical Sciences

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1 International Journal of Biological and Physical Sciences Comparative Analyses on the Effects of Ethanol Leaf-Extracts of Vernonia amygdalina and Azadirachta indica on Lipid Profile in Albino Rats Department of Biochemistry, Ebonyi State University, Faculty of Biological Sciences Abakaliki, Ebonyi State, Nigeria. Article Info: Author(s): History: Received: Accepted Date: Vol 1 (2), pp, September, 13 Corresponding Author: Department of Biochemistry, Ebonyi State University, Faculty of Biological Sciences Abakaliki, Ebonyi State, Nigeria. offorxtianceo@yahoo.com Article Type: Full Length Research Keywords: Ethanol leaf-extracts, Vernonia amygdalina, Azadirachta indica, lipid profile and albino rats Abstract The comparative analyses on the effects of ethanol leaf-extracts of Vernonia amygdalina and Azadirachta indica on lipid profile were conducted using albino rats. The albino rats were grouped into seven (A, B, C, D, E, F and G) containing ten rats per group. Groups A, B, and C were administered Vernonia amygdalina extract by oral intubation at the doses of, and 6 mg/kg body weights respectively for two weeks with corresponding doses of the extract of Azadirachta indica administered to groups D, E and F. Animals in group G served as the control. The serum levels (mg/dl) of total cholesterol, LDL-cholesterol, HDL-cholesterol and triacylglycerol were studied spectrophotometrically. Total cholesterol, triacylglycerol and LDL-cholesterol levels were significantly (p<.5) reduced by both extracts while HDL-cholesterol levels were increased significantly. There were dose-dependent reductions in the body weights of all the animals that received the extracts. The results showed that the leaf-extract of Vernonia amygdalina was more effective as a cardioprotective agent than that of Azadirachta indica. INTRODUCTION Medicinal plants are part and parcel of human society to combat diseases. Azadirachta indica (Neem) and Vernonia amygdalina (Bitter leaf) are well known as versatile medicinal plants having a wide spectrum of biological activities both nutritionally and pharmacologically. Neem is an evergreen tree, cultivated in various parts of the world. Every part of the tree has been used as traditional medicine for house hold remedy against various human ailments from antiquity (Koul et al., 199). Neem has been extensively used in ayurveda and homoeopathic medicine and has become a cynosure of modern medicine. The importance of the neem tree has been recognized by the US National Academy of Sciences which published a report in 1992 entitled Neem a tree for solving global problems. The advancement of neem research has been documented (Schmutterer, 1995). Crude extracts of different parts of neem have been used as traditional medicine for the treatment of various diseases. Neem bark, leaf, root, flower and fruit, together cure blood morbidity, biliary afflictions, itching, skin ulcers, burning sensations and phthisis (Mitra and Patel, 1963). Apart from these common uses, there are several reports on the biological activities of neem and bitter leaf based on modern scientific investigations. The chloroform extract of neem stem bark is effective against carrageenan induced paw edema in rat and mouse ear inflammation. A methanol extract of the leaves exerts antipyretic effect in male rabbits (Okpanyi and Ezeukwu, 1981). The aqueous extract of the leaves also possesses potent immunostimulant activity as evidenced by both humoural and cell - mediated responses (Sen et al., 1992). The hypoglycemic effect of neem was observed with leaf extract and seed oil in normal as well as alloxan induced diabetic rabbits (Khosla et al., ). Aqueous leaf extract from Vernonia amygdalina induces blood sugar lowering effect in rabbits and rats (Akah and Okafor, 1992; Uhegbu and Ogbuehi, 4). An active fraction from hexane extract of neem seed has been found to completely abrogate pregnancy in rodents when given orally up to a concentration of 1% with no apparent side effect (Mukherjee et al., 1999). Neem seed and leaf extracts are effective against malarial parasites. Components of the alcoholic extracts of leaves and seeds are effective against both chloroquine resistant and sensitive strains of malaria parasite (Badam et al., 4). Aqueous leaf extract from Vernonia amygdalina also elicited an enhanced antimalarial effect of chloroquine in mice infected with chloroquine resistant and sensitive Plasmodium berghei strains (Iwalokun, 8). An aqueous extract of Vernonia amygdalina demonstrated a

2 Offor 15 measles virucidal property (Kasim et al., 1). The lipid profile measures total cholesterol, highdensity lipoprotein (HDL), low-density lipoprotein (LDL), very low density lipoprotein (VLDL) and triacylglycerol levels to assess the risk of cardiovascular disease (Robert, 4). Cholesterol is the major sterol in animal tissues and its amphipatic nature enables it to occur in cell membranes (Nelson and Cox, ). It is abundant in the brain, liver, adrenal glands and nervous system. Dietary cholesterol is obtained from animal products. The liver produces sufficient cholesterol in the absence of enough dietary cholesterol for all normal body functions (Shimkin, ). It is carried in the blood in the form of lipoprotein (Poser, 1996). Cholesterol and triacylglycerol are transported in the body in combination with protein to form lipoproteins. High-density lipoprotein is also called good or healthy lipoprotein. It helps to clear cholesterol deposits in the blood vessels left by another blood component called LDL (Robert, 4). The LDL contains mainly cholesterol and when the amount of cholesterol exceeds the amount metabolized by the body, the excess fat floats around in the blood stream and attaches itself to the inner wall of blood vessels to form plaques or a block known as arteriosclerosis. As LDLcholesterol initiates the formation of plaque, it is known as bad cholesterol (Shubhanker and Wolfing, 5). This work was aimed at investigating the comparative effects of ethanol leaf-extracts of Vernonia amygdalina and Azadirachta indica on serum liver enzymes and lipid profile in albino rats. Materials and Methods Fresh leaves of Vernonia amygdalina and Azadirachta indica were collected from Abakaliki in Ebonyi State, Nigeria. Seventy male albino rats were obtained from University of Nigeria, Nsukka. All the chemicals and reagents used were of analytical standard. Extraction of the plant material About 9g of dried and ground Vernonia amygdalina and Azadirachta indica leaves were each soaked into 17ml of ethanol for 24 hours. The solution of the extract was gotten after several squeezing and filtrations into a graduated beaker and allowed to dry under mild sunlight until a semi-solid extract was gotten. Administration of plant extract Extracts of Azadirachta indica at, and 6 mg/kg of body weights were administered for two weeks to animals in groups A, B and C respectively, with corresponding concentrations of Vernonia amygdalina extract given to animals in groups D, E, and F through oral intubation while those in group G served as the control. Blood sample collection. The blood samples were collected on the fifteenth day following the last day of administration through heart puncture into labeled sterile specimen containers. Lipid profile assay Total Cholesterol, Triacylglycerol, HDL- Cholesterol, and LDL -cholesterol assays were done by the methods of Allain and Roschlaw (1979), Quinica, (1978), Brusten et al. (198) and Friedewald et al. (1972) respectively. Data Analysis All the tested parameters were subjected to statistical analysis using t-test. Differences between means were regarded significant at P<.5 (Oyeka, 1996). Results and Discussion There was a general decrease in physical activities of the animals treated with organic extract. This observation could be as a result of an upset in the body metabolism of the animals treated with the extract due to some chemical components of the extract (Datta, 3). Gregor (1997) observed that an ethanol extract of Ocimum basilicum decreased the physical activities of the animals. The extracts from both Azadirachta indica and Vernonia amygdalina leaves decreased the serum total cholesterol and LDL-cholesterol levels significantly Figures 1-5. Crude saponin extract of Vernonia amygdalina leaves reduced serum cholesterol levels in albino rats (Offor et al., 9). Chu () discovered that ethanol extract of Nigella sativa in ten days decreased the cholesterol levels of the animals. Shubhanker and Wolfing (5) showed that an extract from Oryza sativa decreased cholesterol and LDL levels significantly. The decrease could be that the extract inhibits sodium ion efflux leading to higher Na + -Ca 2+ antiporter which prevents the accumulation of Ca 2+ which could have chellated with cholesterol to form plagues (Schneider, 5).Triacylglycerol levels were also decreased. HDL- cholesterol concentrations were significantly (p<.5) elevated by both extracts dosedependently. HDL was raised higher than LDLcholesterol concentrations in rats fed with the extracts. This is due to the decrease in total cholesterol concentration in the animals administered with the extract (Chu, ). The results also showed that the average body weights (g) of the treated animals also decreased significantly. This could be because the extract delays stomach empting leading to more slow absorption of food substance which results to loss of appetite and decrease in the weight (Okafor, 1996). Ethel et al. (1988) found out that both ethanol and ether extracts of fresh leaves of the African marine plant

3 HDL CONCENTRATION (mg/dl) LDL CONCENTRATION (mg/dl) TOTAL CHOLESTEROL CONCENTRATION (mg/dl) 16 Int. J. Biol. Phy. Sci CONTROL Figure 1: Total cholesterol concentrations (mg/dl) in animals that were treated with extracts of Azadirachta indica and Vernonia amygdalina CONTROL Figure 2: LDL-cholesterol concentrations (mg/dl) in animals that were treated with extracts of Azadirachta indica and Vernonia amygdalina CONTROL Figure 3: HDL-cholesterol concentrations (mg/dl) in animals that were treated with extracts of Azadirachta indica and Vernonia amygdalina caused reduction in the average weights of guinea pigs treated with the extracts. The decrease in body weights may be linked to an observed reduction in feed and water intake (Sofowora, 1993).

4 PERCENTAGE CHANGE TRIACYLGLYCEROL CONC. (mg/dl) Offor CONTROL Figure 4: Triacylglycerol concentrations (mg/dl) in animals that were treated with extracts of Azadirachta indica and Vernonia amygdalina..% 15.% 1.% 5.%.% -5.% -1.% -15.% -.% -25.% -3.% 6 Control GROUPS Figure 5: Percentage change in mean body weights of animals that received Azadirachta indica and Vernonia amygdalina extract. Conclusion These observations suggest that the ethanol leaf-extract of Vernonia amygdalina was more effective as a cardioprotective agent than that of Azadirachta indica, but high concentrations of these extracts could be toxic. References Aka PA, Okafor CC (1992). Blood sugar lowering effect of Vernonia amygdalina in an experimental rabbit model. Phytotherapy Res., 6 (3): Allain OP, Roschlaw AF (1979). Biochemical analysis, 3 rd edition. Oxford University Press, New York Badam L, Deolankar RP, Wagh UV (4). In vitro antimalaria activity of neem (Azadirachta Indica) leaf and bark extract. Ind. J. Malariol., 24: Brusten J, Collins CP, Innocent E (198). Determination of HDL level in human Body. J. Sci., 2: Chu FC (). Effect of plant extracts on animal cholesterol. J. Trad. Med.; 13: Datta UC (3). Study of medicinal plants. J. Medical Plants, 12: Ethel AU, Lace FU, Kapchun AC (1988). Medicinal values of tropical plants. J. Pharmacol., 5: Friedewald SP, Denis C, Siegal CP (1972) Method involved in determination of LDL in human body. J. Sci., 3: 6-11, 44. Gregor FT, (1997). Effects of traditional medicines on liver enzymes. J. Trad. Med., 25: Iwalokun BA (8). Enhanced antimalarial effect of chloroquine by aqueous Vernonia amygdalina leaf extract in mice infected with chloroquine resistant and sensitive Plasmodium berghei strains. Afri. Health Sci., 8(1): Kasim LS, Odunkoye OA, Ukpo GE (1). Preliminary investigation of antiviral activities of Vernonia amygdalina and Strunchium sparganophora. J. Pharm. Sci. Pharm. Prac., 7(1): 27. Khosla P, Bhandra S, Singh J, Seth S, Srivastava RK (). Hyperglycaemic effect of neem. Ind. J. Physiol. Pharmacol., 44:

5 18 Int. J. Biol. Phy. Sci. Koul O, Isman MB, Ketkar CM (199). Traditional medicine for household remedy. Canadian J. Bot., 68: Mitra CR, Patel MS (1963). Indian central oilseeds committee. Hyderabad Mukherjee S, Garg S, Talwan GP (1999). Neem - a contraceptive. J. Ethnopharmacol., 67: Nelson DL, Cox MM (). Principles of Biochemistry. 3 rd edition, Worth publishers, New York Offor CE, Nweke FN, Okaka ANC (9). Effect of crude saponin extract from the leaves of Vernonia amygdalina on serum cholesterol level of rats. J. Sci. Technol., 15: Okafor, J.C. (1996). Agroforestry development in the Bosh Okwangwo division of the Cross River National park. J. Aforestation in Nig., 3: Okpanyi SN, Ezeukwu GC (1981). Antipyretic effect of neem product. Planta Medica, 41: Oyeka CA (1996). An introduction to applied statistical methods. 7 th edition, Nobern Avocation Publishing Company, Enugu, Nigeria Poser AC (1996). Extraction of Juice. J. Trad. Med., 5: -25. Quinica A (1978). Method used in determination of triglyceride in human body. J. Sci., 5: 2-9. Robert DO (4). Lipoprotein. J. Lipid Profile, 3: 19-. Schmutterer H (1995). The neem tree: source of unique natural products for integrated pest management, medicine, industry and other purposes. VCH, Weinheim, Germany Schneider BA (5). Coronary heart disease. J. Cholesterol, 5: Sen P, Medinata PK, Ray A (1992). Neem an immunostimulant. Ind. J. Exp. Biol., 12: Shimkin MS (). Acute toxicities of rotenone and mixed pyrethins in mammals. J. Exp. Biol., 34: Shubhanker KA, Wolfing AA (5). Coronary heart disease. J. Hormonal Therapy, 13: Sofowora EA (1993). Medicinal plants and traditional medicine in Africa. Spectrumbooks Ltd, Ibadan, Nigeria. 286 Uhegbu IO, Ogbuehi KJ (4). Effect of aqueous extract of leaves of Vernonia amygdalina on blood glucose, serum albumin and cholesterol levels in diabetic albino rats. Global J. Pure Appl. Sci., 1(1):

Comparative anti-diabetic effects of the ethanol leaf-extracts of Vernonia amygdalina and Azadirachta indica in albino rats

ISSN: 2319-7706 Volume 4 Number 1 (2015) pp. 201-209 http://www.ijcmas.com Original Research Article Comparative anti-diabetic effects of the ethanol leaf-extracts of Vernonia amygdalina and Azadirachta