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HYPOGLYCAEMIC INVESTIGATIONS OF MISTLETOE (LORANTHUS MICRANTHUS) LEAF
EXTRACTS ON DIABETIC RATS
1Eze-Steven, P. E., 2Njoku, O. U., 3Udedi, S., and 4Ogeneh, B.
1Dept. of Biochemistry, Faculty of Natural Sciences, Caritas University, Amorji-Nike, Enugu State, Nigeria
2Dept. of Biochemistry, Faculty of Biological Sciences, University of Nigeria, Nsukka, Enugu State, Nigeria
3Dept. of Applied Biochemistry, Faculty of Applied Natural Sciences, Nnamdi Azikiwe University, Awka, Anambra State, Nigeria
4Dept. of Medical Microbiology, Col ege of Medicine, University of Nigeria, Enugu Campus, Nigeria
This study investigated the hypoglycaemic activities of water extract of mistletoe leaf in the management of diabetes mellitus. Male
Wistar rats were used for this study and they were housed to acclimatize in five different cages according to their groups. Each group
contained four animals. Diabetes was induced in rats in all but groups 4 and 5 following the intravenous injection of alloxan
monohydrate (90mg/kg) dissolved in normal saline through rat tail vein. Group 1 diabetic rats were treated with 600mg/kg body
weight concentration of crude methanol extract of L. micranthus leaves orally. Group 2 diabetic rats were treated with 600mg/kg body
weight concentration of crude water extract of L. micranthus leaves orally. Group 3 diabetic rats were treated with 250mg/kg body
weight concentration of glibenclamide orally. Group 4 diabetic rats were not treated and served as positive control. Rats in Group 5,
which were non-diabetic, received normal saline and served as negative control. The experiments were repeated using different Wistar
rats for groups 1, 2, and 4 for the second, third, and fourth weeks. The results of this study showed that both the methanol and aqueous
extracts of L. micranthus leaves significantly (P<0.05) reduced mean fasting blood sugar concentrations in normal rats. In diabetic rats,
both extracts caused a significant (P<0.05) reduction in serum glucose levels.
Key Words: Hyperglycaemia, mistletoe, Loranthus micranthus, diabetes mellitus, histopathology, glucose, alloxan.
Copy Right, IJISD, 2012.World Research Publications. All rights reserved. INTRODUCTION
shrubs from different genera including the American mistletoe (Phoradendron leucarpum), the European mistletoe (Viscum Diabetes mellitus is a metabolic disorder characterized by album), and the African mistletoe (Loranthus micranthus), hyperglycaemia with lipoprotein abnormalities (Scoppola et which is the European equivalent of Viscum album and the al., 2001). It is one of the main threats to the human health in American equivalent of Phoradendron. Loranthus micranthus the 21st century (Zimmet, 2000). Diabetes mellitus is of two belongs to the family of African bushy plants called types: Type 1 diabetes mellitus or insulin dependent diabetes Loranthaceae (Anderson and Phillipson, 1982 and Newall et mellitus (IDDM) and Type 2 diabetes mellitus or non-insulin dependent diabetes mellitus (NIDDM). This study focused on the use of traditional plant remedies in the management of MATERIALS AND METHODS
blood glucose during diabetes mellitus using animal models. As we reported earlier, Loranthus micranthus leaf extracts indicated antilipidaemic properties (Eze-Steven and Njoku, Male Wistar albino rats of about 8 to 11 weeks old and weight 2010), so we investigated the hypoglycaemic properties of the range of 100 to 200g were used for this study. The rats, aqueous and methanol extracts of African mistletoe (Loranthus obtained from the animal houses of both the Faculty of micranthus). Traditional plant remedies have been in use Biological Sciences and Veterinary Medicine of the University for centuries in the treatment of diabetes, but only a few have of Nigeria, Nsukka, Nigeria (UNN), were kept under standard been scientifically evaluated (Akhtar and Ali, 1984). In conditions for 7 days with water and food ad libitum for Nigeria, local herbal practitioners claim that extracts from acclimatization before the experiments commenced. mistletoe leaf is effective in the management of high blood glucose levels that is associated with diabetes mellitus Animal Stock
(Osadebe et al., 2004). Mistletoe is a semi-parasitic woody perennial plant commonly found growing on oaks and other Fifty-six male Wistar rats were used for this study. They were deciduous trees. Its semi-parasitic nature is because the plant acclimatized and housed in separate cages according to their synthesizes its own chlorophyll but depends on the host for its groups. Diabetes was induced in rats by the intravenous supply of water and minerals. (Duke, 1985). The plant relates injection of alloxan monohydrate (90mg/kg) through the rat’s to several different species of perennial, evergreen, parasitic tail vein. Diabetic condition in the rats was observed with an increase in the rat’s blood glucose concentration and other *Corresponding author: email@example.com
clinical features including loss of weight, increased frequency 016
International Journal of Innovation Sciences and Discoveries, Vol. 1, No, 01, pp. 015-017, November, 2012
Following the induction of diabetes, the rats were divided into blood sample. The colour intensity was read instantly from the five groups of four animals each (n = 4). Group 1 was diabetic rats treated with 600mg/kg methanol
extract (ME) of Loranthus micranthus orally. Group 2 was diabetic rats treated with 600mg/kg of the
The results of the experiment were presented as meanSEM aqueous (distilled water) extract orally and was called the AE. and were subjected to One Way Analysis of Variance Group 3 was diabetic rats treated with 250mg/kg of
(ANOVA). The differences between the means were tested glibenclamide orally. Diabetic untreated rats named the using post Hoc L.SD at P0.05 significance level. Group 4. And Group 5 included non-diabetic rats given
The methanol and aqueous extracts significantly (P<0.05) The experiments were repeated with different animals for reduced the fasting blood sugar in normal wistar rats. The groups 1, 2, and 3 for the second, third, and fourth weeks. reduction in fasting blood sugar caused by the methanol extract was higher within the first 6 h post-administration compared to PLANT MATERIAL
that of the aqueous extract. The effect of the aqueous extract increased gradually within the first 2 h post-administration. The experiment was carried out in two parts with the same Both extracts did not significantly (P>0.05) affect the fasting plant material. One part was carried out using methanol extract blood sugar in normal rats after 18 h post-administration. while the other was carried out using aqueous (distilled water) extract.
Preparation of the Methanol and Aqueous Extracts
The procedure for the preparation of the methanol extract was
as we earlier reported (Eze-Steven and Njoku, 2010). The
second part of the dried leaves were also pulverised into coarse
form and used for the preparation of the aqueous extract.
Determination of the Concentration of Various Extracts
To determine this, known weights of both extracts were
determined separately. The weight of dry crucible was also Fig. 1: Effect of extracts on fasting blood sugar concentration of
determined. Later, known weights of both extract were put into normal rats
the dry crucible, respectively, and their weight determined before heating. The crucible with its content was heated and
after the heating, the crucible was weighed with its heated
content and the weight recorded. The concentrations of both
extracts were then calculated from the various weights.
Acute Toxicity Test (LD50)
The median lethal doses (LD50) of the methanol extract and
aqueous extract were determined in mice using the oral route
of administration (Lorke, 1983). Both extracts did not cause
death in mice at dose concentrations above 2900mg/kg.
Consequently, the methanol extract and aqueous extract of
mistletoe (Loranthus micranthus) are considered non toxic
(Lorke, 1983) at such concentration.
Determination of Plasma Glucose concentration
Fig. 2: Effect of extracts on blood glucose level of diabetic rats
ONE-TOUCH blood glucose monitoring meter and test strips The methanol and aqueous extracts significantly (P<0.05) reduced the blood glucose level of alloxan-induced diabetic California, USA) were used for this assay. The principle of the rats within the four weeks of treatment. The extracts caused reaction is based on the glucose oxidase reaction. The blood highest reduction in serum glucose level after the second week glucose estimation involves the reaction between glucose in of treatment. The magnitude of effect caused by the methanol the blood and oxygen in the presence of glucose oxidase, extract was significantly (P<0.05) higher than that of the which is immobilised in the test strip, to yield gluconic acid and hydrogen peroxide. The hydrogen peroxide subsequently oxidizes the dye in a reaction mediated by peroxidase to DISCUSSION AND CONCLUSION
produce a blue-coloured product. The intensity of the colour produced is proportional to the glucose concentration in the The results of this research work show a gradual reduction in the serum glucose concentrations in both fasting and non- 017
International Journal of Innovation Sciences and Discoveries, Vol. 1, No, 01, pp. 015-017, November, 2012
fasting states in the diabetic and non-diabetic rats. The crude Duke, J. A. 1985. CRC Handbook of Medicinal Herbs. Boca extracts gradually reduced the serum fasting glucose concentrations in the non-diabetic rats. In the diabetic states, Eze-Steven, P. E. and Njoku, O. U. 2010. Antilipidaemic both extracts also indicated an antihyperglycaemic activity. studies of mistletoe (Loranthus micranthus) leaf extracts This shows that the methanol extract and aqueous extract of on diabetic rats. Int J Curr Research. 8: 048-055. Loranthus micranthus have antihyperglycaemic properties Hoglund, P. 1999. Initiation of autoimmune diabetes by with the methanol extract having a greater effect than the developmentally regulated penetration of islet cell aqueous extract. This result confirms that the methanol extract antigens in the pancreatic lymph nodes. J Exp Med. 189: shows antilipidaemic properties as we reported earlier but might have some destructive effect on the hepatocytes of Lorke, D. 1983. A new approach to practical toxicity testing. treated rats. This could be a result of destructive effect of the Archives of Toxicology. 54: 275-287. alcohol on the hepatocytes of treated rats. It further indicates Newall, C. A., Anderson, L. A. and Phillipson, J. D. 1996. that the methanol extract has a less destructive effect on the Herbal medicines: a guide for health-care professionals. kidneys than it has on the liver. The liver is a vital organ involved in the detoxification and deamination reactions in the Osadebe, J. O., Okide, G. B. and Akabogu, I. C. 2004. Study body. The kidneys are involved in the excretion of metabolites, on anti-diabetic activities of crude methanolic extracts of mostly inactive, from the body. Thus, during the process of Loranthus micranthus (Linn.) sourced from five different host trees. J Ethnopharmacol. 95: 133-138. compounds in the methanol extract destroyed the hepatocytes Ross and Wilson. (2001). Anatomy and Physiology in Health of the organ of detoxification more than those of the kidney and illness. Churchill Livingstone Pub., UK. pp. 234. Scoppola, A., Montecchi, F. R., Mezinger, G. and Lala, A. 2001. Urinary mevalonate excretion rate in type 2 REFERENCES
diabetes: role of mevalonate control. Artheroscl. 1556: Akhtar M. S. and Ali M. R. 1984. Study of antidiabetic effect Zimmet, P., Alberti, K. G. M. M., and Shaw, J. 2001. Global of a compound medicinal plant prescription in normal and and societal implications of the diabetes epidemic. Nature. diabetic rabbits. J Pakistan Med Assoc. 34: 239-244. Anderson, L. A. and Phillipson, J. D. 1982. Mistletoe – the magic herb. Pharm J. 229: 437 – 439. Brownlee, M. (2001). Biochemistry and Molecular cell biology of diabetic complications. Nature. 414: 281-320.
The item below was posted on journalreview.org on May 1, 2007, attached to the article by Vaccarino et al., but discussing article by Jha et al and Trivedi et al. that appeared with the Vaccarino article in the August 28, 2005 issue of the New England Journal of Medicine. In light of the closing of the journalreview.org site, it is reproduced here. A correction regarding the statements concer