Bajopas Volume 9 Number 1 June, 2016

http://dx.doi.org/10.4314/bajopas.v9i1.21

Bayero Journal of Pure and Applied Sciences, 9(1): 138 - 141

Received: January, 2016
Accepted: April, 2016
ISSN 2006 – 6996

ANTIDIABETIC EFFECT OF FERMENTED Pennisetum glaucum (MILLET)

SUPPLEMENT IN ALLOXAN INDUCED HYPERGLYCEMIC WISTAR RATS
*Sada, N.H., Tanko, Y., Gidado, N.M. and Mohammed, A.
Department of Human Physiology, Ahmadu Bello University, Zaria, Nigeria.
* Corresponding author: Email-yusuftanko@yahoo.com
ABSTRACT
Diabetes mellitus is a debilitating disease that is characterized clinically by hyperglycemia due to
chronic and/or relative insulin insufficiency. The disease is associated with disturbance in lipid and
protein metabolism. Pennisetum glaucum (PG) has been recommended for several therapeutic
purposes as it has been shown to have high amount of magnesium which helps to increase the
levels of adiponectin hormone but the hypoglycemic effect of millet as a supplement is yet to be
fully elucidated. Therefore this research was designed to determine the effect of Pennisetum
glaucum supplement on blood glucose level and serum lipid profile. Diabetes mellitus was induced
by single intraperitoneal (i.p) injection of alloxan dissolved in 0.1ml fresh cold citrate buffer pH 4.5
at a dose of 150 mg/kg body weight, after which the rats were randomly divided into 5 groups.
Group 1 normoglycemic, Group 2 Diabetic untreated, Group 3 Diabetic treated with 1 mg/kg of
Glibenclamide (GB). Group 4 Diabetic treated with 33%w/v P.G. supplement, Group 5 Diabetic
treated with 66% P.G. supplement. When compared with the diabetic control, the study revealed a
significant decrease (p>0.05) in blood glucose level at both 33%w/v and 66% w/v P.G.
supplementation. It also showed that P.G. at both 33% and 66% supplement was able to lower
Triglycerides and LDL serum levels as well increase HDL serum levels although not statistically
significant, but significantly restored HDL/Cholesterol and Triglycerides/HDL ratios to normal
physiological range. Pennisetum glaucum supplementation showed high hypoglycemic effect and
also hypolipidemic property in alloxan induced hyperglycemic wistar rats.
Keywords: Pennisetum glaucum, hypolipidemia, hyperglycemia
INTRODUCTION resulting in individuals with this condition relying
Diabetes is a serious health hazard currently affecting essential on exogenous insulin administration for
more than 220 million people worldwide and survival, although a subgroup has significant residual
expected to afflict 400 million by 2030 (Wild, C-peptide production (Keenan et al., 2010). So far,
2004; Shaw et al., 2010). The greatest increase in the management of this disorder entails increased
prevalence in the near future, however, is expected to physical activity, healthy eating or diet and
occur in Asia, the Middle East, and Africa, where it is administration of anti-diabetic drugs and/or insulin.
likely that there will be a 50% increase in diabetes in However, the currently available anti-diabetic drugs
these parts of the world (Shaw et al., 2010). This is of are have numerous side effects. Besides, these agents
public health concern due to its social and economic are costly and, in some cases, not readily available
burdens. Eventhough diabetes has no known cause, (Omotayo, 2014).
complex interplay of several factors including genetic, Pennisetum glaucum, also known as Pearl Millet is
social, and environmental factors are implicated in its a cereal crop grown in tropical semi-arid regions
etiology. It is a metabolic disorder characterized by of the world primarily in Africa and Asia. Pearl
insufficient secretion of insulin or resistance to the millet is rich in several nutrients as well as non-
action of insulin or both. The major clinical nutrients such as phenols. It has high energy, less
manifestation of type 1 diabetes state is starch, high fiber, low glycemic index and gluten
hyperglycemia and hypoinsulinemia however, insulin free. The protein content ranges from 8 to 19% and it
deficiency and/or insulin resistance also are is low in lysine, high in tryptophan, threonine and the
associated with disturbance in lipid and protein sulfur-containing amino acids. Pearl millet is rich in fat
metabolism (Mutaa and Omar, 2013). In type 1 content (NIN 2003) such as Omega 3, linolenic acid
diabetes, hyperglycemia occurs as a result of a (C18:3n-3) (LNA) comprises 4% (Burton et al, 1972).
complex disease process where genetic and It contains various essential micro nutrients needed
environmental factors lead to an autoimmune by the body. Its mineral content are B-vitamins,
response that remains to be fully elucidated. During potassium, phosphorous, magnesium, iron, zinc,
this process, there is the destruction of the copper and manganese (NIN, 2003).
pancreatic-cells within the islets of Langerhans

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It has been recommended for several therapeutic pellets grower mash only , Group II (diabetic control)-
purposes, as it has been found to inhibit tumor Alloxan induced hyperglycemic wistar rats +normal
development (Lee, 2010), control blood pressure and saline (0.2ml) orally fed with, Group III- Alloxan
plasma low density lipoprotein cholesterol levels induced hypeglycemic wistar rats + 33% millet
(Hosoda, 2012), and possesses anti-allergenic supplement (Goodarzi, et al., 2011), Group IV-
characteristics. Alloxan induced hyperglycemic wistar rats + 66%
The aim of the study was to evaluate the effect of the millet supplement (Goodarzi, et al., 2011), Group V-
Pennisetum Glaucum supplement on blood glucose Alloxan induced hyperglycemic wistar rats +
level and lipid profile in alloxan induced hyperglycemic glibenclamide (5mg/kg) orally
wistar rats. Blood Glucose Determination
Fasting blood glucose levels was determined at day 0,
MATERIALS AND METHODS 4, 8, 12 and 14 respectively for 2weeks using the
Experimental Animals glucose oxidase principle (Beach and Turner, 1958)
A total of 25 wistar rats of both sexes weighing 160g- using one touch digital glucometer
200g were purchased at the experimental animal Sample Collection
house of the Department of Human Physiology, At the end of the experiment, after overnight fasting,
Ahmadu Bello University Zaria. Animals were the animals were anaesthesized using chloroform and
maintained under normal laboratory conditions. The dissected, blood was collected through cardiac
animals were fed on pellets of growers mash and puncture, centrifuged and serum was used for
allowed access to water ad libitum. The study was assessment of lipid profile.
conducted in accordance with the guidelines of Lipid Profile Assay
Ahmadu Bello University in accordance with the rules The method of Roeschlau and Gruber was applied
governing the use of laboratory animals as accepted using assay kits (Randox Laboratories Ltd., UK) to
internationally (National Institute of Health Guide for determine the total serum cholesterol
Care and Use of Laboratory Animals) spectrophotometrically at 546 nm following enzymatic
Collection, Identification and Preparation of hydrolysis and oxidation (Allain et al., 1974). The
Supplement HDL-cholesterol was determined using assay kits
Pennisetum Glaucum was purchased from Institute of (Randox Laboratories Ltd., UK) after low density
Agricultural Research Zaria, Kaduna State of Northern lipoprotein (LDL and VLDL) and chylomicron fractions
Nigeria. It was identified at the Herbarium unit of the were precipitated quantitatively by the addition of
department of Biological Sciences, Ahmadu Bello phosphotungstic acid in the presence of magnesium
University, Zaria and a specimen voucher with the ions. After centrifugation, the cholesterol
number 27111 was deposited. Three (3) kg of the concentration in the HDL fraction in the supernatant
pearl millet (Pennisetum Glaucum ) was soaked in was assayed colorimetrically at 540 nm (Allain et al.,
water for 72 hours. Thereafter, it was air dried under 1974).
the sun for seven days. Later it was grounded to Statistical Analysis
powder using grinding machine. Data collected were expressed as mean + SEM. They
Drugs Used were analyzed by one way analysis of variance,
Alloxan was purchased from (Sigma chemical ANOVA, and Tukey’s post-hoc test was used to
company St. Louis U.S.A), glibenclamide and all other compare the level of significance between the control
chemicals and reagents that were used were of and treatment groups, using SPSS version 20.0.
analytical grade. Values of p<0.05 were considered significant.
Induction of Diabetes Mellitus
After 16-18 hours of fasting, hyperglycemia was RESULTS
induced by a single intraperitoneal injection of alloxan Effect of Pennisetum Glaucum (PG) supplement
monohydrate ( Sigma St. Louis, M.S., U.S.A.) at a on blood glucose level: The results of the present
dose of 150mg/kg body weight (dissolved in 0.9% study showed that blood plasma glucose level in the
NaCl cold normal saline solution) ( Katsumata et al., diabetic control group was significantly
1999). Rats were treated with 20% glucose solution higher(p<0.05) when compared with the normal
orally for 6hrs due to the ability of alloxan to produce control group in the entire 16 days of the study. The
fatal hypoglycemia as a result of massive pancreatic effect of P.G. on blood plasma glucose level was
insulin release. They were placed on 5% glucose determined by comparing the values in the diabetic
solution for 24 hrs to prevent hypoglycemia control group (i.e. Group 2) with the values in Group
(Dhandapani et al., 2002). Blood was collected from 4 (i.e. 33% P.G.) and in Group 5 (i.e. 66% P.G.).
the tail vein of the rats after 72hrs of alloxan These results showed that P.G. supplement was
injection. Rats having fasting blood glucose level lowered the blood Glucose Level (p<0.05) within
greater than 200mg/dl were selected for the study sixteen days of its administration dose dependently on
(Matteucci and Giampietro, 2008). Day 12 and 16 respectively, when compared with the
Experimental Design and Treatment Diabetic Control. Its effect appeared to be even more
Wistar rats were weighed and randomly assigned into potent and significant than that of the standard drug
5 groups, with 5 animals (n=5), in each. Group 1 glibenclamide (Table 1)
(normal control) - Normogycemic wistar rats fed with

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Table 1: Effect of Sixteen Days Administration of Pennisetum glaucum (PG) Supplement on Blood
Glucose Levels (mg/dl) in Alloxan Induced Hyperglycemic Wistar Rats
Group/Treatment Day 0 Day 4 Day 8 Day 12 Day 16
Normal control 68.00 ± 6.25 50.40 ± 2.75 70.60 ± 7.09 63.80 ± 3.87 70.40 ± 3.54
Diabetic control 430.60 ± 56.14 414.60 ± 80.37 326.40 ± 31.04 324.20 ± 23.06 292.40 ± 25.13
33% PG 434.60 ± 53.22 456.80 ± 88.31 315.20 ± 58.20 253.00 ± 70.79 128.80 ± 34.50*
66%PG 434.20 ± 69.98 418.00 ± 74.35 246.40 ± 50.70 143.60 ± 26.26* 106.40 ± 15.48*
Glibenclimide 430.00 ± 74.65 332.40 ± 72.29 181.80 ± 38.94 120.20 ± 27.43* 85.40 ± 19.03*
Group 1: Normoglycemic Rats; Group 2: Hyperglycemic Rats (Diabetic Control); Group 3: Diabetic Rats +
2mg/kg Glibenclamide (Positive Control); Group 4: Diabetic Rats Treated with 33% of P.G. Supplement; Group 5:
Diabetic Rats Treated with 66% of P.G. Supplement.
* Statistically Significant at p< 0.05 when compared with Untreated Diabetic Control

Effect of Pennisetum Glaucum Supplement on glibenclamide. It also showed that P.G. at both 33%
Serum Lipid Profile: These results of the present and 66% supplement lowered the Triglycerides and
study revealed that P.G. at 33% and 66% supplement LDL serum levels( p<0.05) as well increase HDL
was able to lower cholesterol levels dose dependently serum levels although not statistically significant
and significantly when compared to the diabetic (Table 2).
untreated group more than the standard drug

Table 2: Effect of Sixteen Days Administration of Pennisetum glaucum (PG) Supplement on Lipid
Profile in Alloxan Induced hyperglycemic Wistar Rats
Group/Treatment Total Cholesterol(mmol/L) Triglycerides(mmol/L) HDL(mmol/L) LDL(mmol/L)
Normal control 1.16 ± 0.10 1.11 ± 0.10 0.51 ± 0.09 0.14 ± 0.04
Diabetic control 2.64 ± 0.08 1.42 ± 0.12 0.33 ± 0.07 1.46 ± 0.14
33% PG 1.72 ± 0.26 0.81 ± 0.01 1.85 ± 0.81 0.10 ± 0.99
66%PG 1.60 ± 0.35* 1.39 ± 0.14 0.60 ± 0.21 0.43 ± 0.26
Glibenclimide 1.43 ± 0.25* 1.21 ± 0.25 0.65 ± 0.14 0.23 ± 0.08
Group 1: Normoglycemic Rats; Group 2: Hyperglycemic Rats (Diabetic Control); Group 3: Diabetic Rats +
2mg/kg Glibenclamide (Positive Control); Group 4: Diabetic Rats Treated with 33% of P.G. Supplement; Group 5:
Diabetic Rats Treated with 66% of P.G. Supplement.
* Statistically Significant at p< 0.05 when compared with Untreated Diabetic Control

DISCUSSION yeast has shown to possess hypoglycaemic

The results of the present study showed that blood property that is comparable to the oral
plasma glucose level in the diabetic control group was hypoglycaemic drug, glibenclamide as described by
significantly higher(p<0.05) when compared with the Mohammed et al. (2015). Results of lipid profile are
normal control group in the entire 16 days of the considered as good indicators of whether someone is
study. The effect of P.G. on blood plasma glucose prone to develop stroke or heart attack, caused by
level was determined by comparing the values in the atherosclerosis. Hyperlipidaemia, a risk factor of
diabetic control group (i.e. Group 2) with the values in cardiovascular diseases is frequently seen among
Group 4 (i.e. 33% P.G.) and in Group 5 (i.e. 66% diabetic patients. Serum lipid levels are commonly
P.G.). These results showed that P.G. supplement was increased in diabetes mellitus and such an elevation
able to lower blood glucose Level within sixteen days represents a risk factor for coronary heart disease
of its administration dose dependently and (Daniel et al., 2011). The results of the present study
significantly particularly on Day 12 and Day 16 when revealed that P.G. at 33% and 66% supplement was
compared with the Diabetic Control. Its effect able to lower cholesterol levels dose dependently and
appeared to be even more potent and significant than significantly when compared to the diabetic untreated
that of the standard drug glibenclamide. group more than the standard drug glibenclamide. It
Glibenclamide, a well-known oral hypoglycaemic also showed that P.G. at both 33% and 66%
drug belongs to the family of the sulfonylureas a supplement was able to lower Triglycerides and LDL
first generation anti-diabetic drug; elicits its serum levels as well increase HDL serum levels
hypoglycaemic effect mainly by acting on ATP- although not statistically significant, but significantly
sensitive potassium channel in pancreatic beta-cells. restored HDL/Cholesterol and Triglycerides/HDL ratios
The blood glucose lowering effect of P.G. and the to normal physiological range. The findings of the
possible mechanism involved in the hypoglycemic present study agrees with the findings of Lee(2010)
action of P.G. may be stimulation of insulin secretion who reported that the levels of triglycerides, low
by the pancreas or/and enhance insulin sensitivity in density lipoprotein decreased while high density
various organs due to its high amount of magnesium lipoprotein levels increased significantly in diabetic
and selenium. Magnesium helps to improve the ability mice fed P.G. supplement (Lee, 2010). Medugu et al.,
of cells to respond to insulin, by increasing the levels (2010) showed that abdominal fat of broilers on
of adiponectin hormone. Adiponectin prevents the millet-based diets were significantly lower than those
accumulation of glucose in the blood by increasing fat on maize and high-tannin sorghum based diets.
metabolism in tissues (Nishizara, 2009). Selenium-
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These indicate that millet and sorghum can be well- cholesterol lowering effect of P.G. may be due to the
utilized to produce broiler chickens with superior carcass availability of flavonoids in it, Flavonoids as antioxidants
quality compared to maize. Nishizara also showed that may inhibit the oxidation of LDL cholesterol (prevention
dietry Japanese millet protein ameliorated plasma levels of atherosclerotic plaque formation) (Middleton, et al.,
of lipids in type 2 diabetic mice and also increased the 2000; Duarte-Alameida, et al., 2007).
levels of high density lipoprotein (Nishizara, 2009). CONCLUSION
The ability of P.G. supplement to reduce plasma The study confirmed the association between of
cholesterol and triglycerides in diabetic animals could be hyperglycemia and abnormal lipid profile. Administration
explained by its reported high content of magnesium. of Pennisetum glaucum supplement at different
Magnesium helps to improve the ability of cells to percentages lowered blood glucose level which was even
respond to insulin, by increasing the levels of adiponectin more effective than the standard drug glibenclamide.
hormone. Adiponectin is produced in fat tissues, which is Pennisetum glaucum lower the cholesterol levels more
a beneficial hormone that helps in energy metabolism, it than the standard drug glibenclamide. Its lower
helps cells burn fats for energy and improve triglyceride levels, increased serum HDL levels although
cardiovascular health (Nishizara, 2009). Since insulin is not statistically significant, but significantly restored
required for the inhibition of hormone-sensitive lipase HDL/Cholesterol and Triglycerides/HDL ratios to normal
and hence increases the utilization of glucose and physiological range. Hence, it can be concluded that
thereby decreasing the mobilization of free fatty acids supplementation with Pennisetum Glaucum has high
from the fat depots. The decreased level of FFA is also hypoglycemic effect and also hypolipidemic property in
associated with decreased actions of lipolytic hormones, alloxan induced hyperglycemic wistar rats.
which, in turn, decreased the activity of hormone
sensitive lipases on fat deposits (Daniel et al., 2015). The

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