Effect of Black Rice Bran Extract to Decrease Glucose Level of

Diabetic Rats
Novita Sari* and Arifah Sri Wahyuni
Fakultas Farmasi Universitas Muhammadiyah Surakarta
Jl. A. Yani Tromol Pos I, Pabelan Kartasura Surakarta 57102
*e-mail: [email protected]

Abstract
Black rice bran is one of processed foods derived from plants and has a decreasing effect of blood glucose
levels. Major component of anthocyanin in black rice bran that functions in lowering blood glucose
concentration is cyanidin 3-glucoside. Fifteen male rats Sprague-Dawley strain were selected randomly and
divided into 5 groups with the same number in each groups. These groups were consisted of normal control,
negative control, and black rice bran extract treatment with dose of 50, 100 and 200 mg/kg of bodyweight
(b.w.). Each of them were induced to experience diabetes by alloxan 150 mg/kg of b.w. Measurement of
blood glucose levels was done by using visible spectrophotometer at 546 nm. The samples were collected
before and after alloxan injection, and after 4, 7, and 10 days later to observe the changes of blood glucose
levels. The results showed that black rice bran at 200 mg/kg of b.w. was able to lower the glucose levels,
which amounted for 131,33 ± 8,08 mg/dL after 10 days of injection.

Keywords: Black rice bran, anthocyanin, antidiabetic.

Introduction been developed optimally. Black rice bran has

Diabetes mellitus (DM) is one of the a main component of cyanidin 3-glucoside as
health problems caused by the degenerative the active compound to lower blood glucose
process. Various epidemiological studies have levels (Park et al., 2008). In a previous research
shown that an increased prevalence of diabetes conducted by Hadi (2015), the accumulation
mellitus (Suyono, 1995). Global Prevalance of anthocyanins in black rice bran amounted
Of Diabetic predicts an increase of diabetes in to 3.284 mg/100g, whereas research
the coming years with the prevalence of the conducted by Suryaningrum (2015) levels of
growth was two-fold in developing countries. cyanidin 3-O-glucoside in the ethanol extract
In Indonesia, it is estimated that the number of of black rice bran was 0.012% (w/w) more
patients will rise up to 21.3 million in 2030, than the ethanol extract of red rice bran. This
which was previously only about 8.4 million compound with 200 mg/kg dose was able to
in 2000 (Wild et al., 2004). As the amount of reduce blood glucose concentration in the
people with DM gaining every year, it becomes diabetic nephropathy rat equivalent to normal
pharmacist duty to educate and prevent this controls (Nawa 2014). Flavonoid compounds
disease in the community. in black rice brand have a mechanism of
The accumulation of patients with DM action in reducing blood glucose levels by
annually causes upsurge of antidiabetic drugs stimu-lating insulin secretion and increasing
consumption, even though the use of synthetic Ca2+ influx in the tissues (Tapas et al., 2008).
antihyperglycemic drugs can cause adverse Flavonoids are able to prompt glucose uptake
side effect. Nowadays, traditional herbal in tissue and regulate the level of enzyme
medicines with minimal detrimental side activity through the carbohydrates metabolism
effect has been developed. One of these drugs (Brahmachari, 2001).
comes from plants that have the potential to The purpose of this research was to
lower blood glucose levels (Malviya et al., determine the effect of black rice bran extracts
2010). One of the foods that come from plants on the reduction of blood glucose levels in
and has this ability is black rice bran. diabetic rats. The results of the study were
The use of black rice bran as an expected to provide information about the
antihyperglycemic drug in Indonesia has not benefit of black rice bran extract in reducing

8 PHARMACON, Volume 18, Juni 2017

blood glucose levels. Hence, it can be used as mice were randomly selected and divided into
an alternative antidiabetic herbal medicine. 5 groups with each consisting of 3 rats

Research Methodology - Group I (normal control), fed with pellets

1. Experimental Design and distilled water po
This research was an experimental - Group II (negative control) was treated
study with completely randomized design to with alloxan and given distilled water as
determine the effect of black rice bran extract a po
in reducing blood glucose levels in diabetic - Group III was treated with an extract of
rats. The tests were carried out by using 15 50 mg / kg po
white Sprague-Dawley strain male rats which - Group IV was treated with an extract of
were divided into five treatment groups. This 100 mg / kg po
study was conducted at the Laboratory of - Group V was treated with an extract of
Pharmacology and Clinical Pharmacy Faculty 200 mg / kg po
of Pharmacy, Universitas Muhammadiyah
Surakarta. Rats were weighed every day until
the 10th day as a baseline. They were fasted
2. Tools and materials for 8-12 hours on the day before the initial
a. Tools used in the extraction process measurement of blood glucose levels (called
were macerator, rotary analytical day 0). Next, group II-V were induced with
balance, evaporator, filter paper, alloxan 150 mg/kg of b.w. intraperitoneally.
Buchner funnel and vacuum pumps, Monitoring of the blood glucose levels was
aluminum foil, porcelain bowls, and done until the 2nd day after the induction
water bath. of alloxan. Rats were considered to have
b. Tools used for animals testing were diabetes if the concentration of blood glucose
rat cages, scales, drinking places, was more than 140 mg/dl. The test animals
gloves, masks, centrifuge, vortex, that already have experienced diabetes can
oral sonde, needles, and disposable be treated with extracts. Analysis of blood
syringes. glucose concentration were performed on the
c. Tools used for blood glucose levels 4th, 7th, and 10th day.
measurements were disposable
semi-micro cuvette, micropipette 5. Determination of Glucose
size from 0.5 to 10 mL and 100- Concentration
1000 mL (Accura), white tips, blue As much as 0.5 ml of blood sample
tips, and UV-Vis spectrophotometer was taken from lateral tail vein and put
(Star Dust FC * 15), pipette, into microtube. Then, these samples were
Beker glass, cuvette and UV-Vis centrifuged at 13.400 rpm for 30 minutes to
spectrophotometer. obtain the blood serum, which will be used as
samples for analysis.
3. Materials There were blank, standard, and sample
Materials used of this research were black solution used in this analysis. All solutions
rice bran extract, male rats Sprague-Dawley were made by adding 5 ml aquadest, glucose,
strain weighing 200-300 grams, and reagent and blood serum to 500 ml of reagent Glucose
Glucose GOD FS kit. GOD FS kit respectively. Then, these mixtures
were vortexed and incubated for 10 minutes
4. Treatment for Laboratory Animals at 37°C. The absorbance was read by using
Test animals in this study were male rats spectrophotometer at a wavelength of 546
2-3 months old with a weight of 200 to 300 nm. The data acquired were analyzed by using
grams that have acclimatized for 7 days. Then, statistical MS Exel worksheet.

PHARMACON, Volume 18, Juni 2017 9

Results And Discussion no decrease in the blood glucose levels in this
1. Effect of Black Rice Bran Extracts group. On the other hand, the negative control
Toward Decrease of Blood Glucose group did not receive any additional treatment
Level. after alloxan adminis-tration. Consequently,
Regulation of blood glucose are the negative control group did not show
performed by pancreas by releasing insulin. decrease in blood glucose levels, which
Therefore, pancreatic damage will result in amounted for 295.33 ± 157.27 mg / dL (Figure
decline of insulin secretion (Chambpel et 1). Based on the results of Wilcoxon statistical
al., 2004). Defect in the pancreatic cells can test, black rice bran extract treatment doses of
be induced by alloxan. It is a diabetogenic 50, 100 and 200 mg/kg of bw can reduce blood
agent, which has the chemical formula glucose levels significantly (p<0.05). This
2,4,5,6-pyrimidine trione and unstable in the decline was happened in the 4th, 7th, and 10th
water. It has a half life of 1.5 minutes at neutral day after treatment. The largest reduction was
pH at 37 °C and produce dialuric acid through occurred on 200 mg/kg of bw black rice bran
redox cycling process that will form peroxide extract which accounted for 131.33 ± 8.08 mg/
radicals. Through the Fenton reaction, reactive dL after 10th day of treatment (Table 1).
hydroxyl radicals will increase calcium ions Flavonoid compounds contained in black
level and cause damage to the β-Langerhans rice bran, which is called anthocyanins (Park et
cells (Szkuldeski, 2011). Alloxan dose of 150 al., 2008), caused a decrease in blood glucose
mg/kg proved to increase the average level concentration. The major anthocyanin
of blood glucose up to 300 mg/kg (Table 1).
According to Etuk & Muhammed (2010), this
dosage was shown to increase the glucose
concentration in the 75% population of test
animals.
The observation of changes in blood
glucose levels were performed on the fourth,
7th, and 10th day after the extract treatment. The
treatment groups in this study were using 50,
100, and 200 mg/kg of b.w. of black rice bran
extract. These dosage levels were based on
Nawa (2014) which stated that these dosages
were able to decrease the blood glucose levels.
The treatment control group was fed only Figure 1. Average of glucose level (mg/dL) before
and after treatment extract.
with pellets and water. Therefore, there was

Table 1. Glucose level (mg/dL) before and after treatment extract on 0, 4, and 7 days.
Glucose level (mg/dL)
Group
Baseline Days 0 Days 4 Days 7 Days 10
83 83 90 161 126
Normal group 92 116 112 122 149
155 152 88 136 152
X ± SD 110 ± 39,23 117 ± 34,51 96,67± 13,31 139,67 ± 19,75 142,3 ± 14,22
II 190 230 545 469 447
Negative group 120 206 137 147 133
132 519 457 418 306
X ± SD 147,33 ± 37,43 318,33 ± 174,19 397,67 ± 214,71 344,67 ± 173,07 295,33 ± 157,27
III 134 414 475 193 345

10 PHARMACON, Volume 18, Juni 2017

 Glucose level (mg/dL)
Group
Baseline Days 0 Days 4 Days 7 Days 10
Extract BBH 50mg/kgBB 100 669 637 547 583
116 243 195 184 169
X ± SD 116,67 ± 17,00 442 ± 214,37 451,67 ± 198,03 308 ± 207,02 365,67 ± 207,77
IV 138 240 214 141 124
Extract BBH 100mg/kgBB 181 474 154 218 152
149 486 430 184 141
X ± SD 156 ± 22,33 400 ± 138,69 266 ± 145,61 151 ± 38,58 139 ± 14,10
V 118 347 213 108 130
Extract BBH 200mg/kgBB 136 341 167 122 124
204 686 431 174 140
X ± SD 152,67 ± 45,35 458 ± 197,47 270,33 ± 141,02 134,67 ± 34,77 131,33 ± 8,08

compounds that has activity in lowering glucose uptake into peripheral tissues and
blood glucose levels is cyanidine 3-glucoside regulating the level of enzymatic activity that
(Kaneda et al., 2006). In a study conducted by responsible in coordinating the metabolic
Suryaningrum (2015), cyanidine 3-glucoside pathways (Brahmachari, 2011).
obtained from ethanol extract of black rice In addition, Kaneda et al., (2006)
bran as much as 0.012% w/w. Moreover, Hadi explained that flavonoid anthocyanin also act
(2015) concluded that black rice bran has a as an antioxidant which is important in the
total anthocyanin content of 3.284 mg/100 g. inhibition of the formation of Reactive Oxygen
The presence of these compounds can improve Species (ROS) by protecting pancreatic beta
the situation of hyperglycemia by increasing cells while simultaneously reducing oxidative
glucose transporter type 4 (GLUT4) protein stress in diabetes melitus (Park et al., 2008;
to facilitate the entry of glucose into the cells Modak et al., 2007). Thus, antioxidants in
(Sasaki et al., 2007). Hence, flavonoids have black rice bran can prevent complications of
similar ability with insulin in mode of action diabetes mellitus (Rahimi et al., 2005).
and repairing pancreatic beta cells. According Conclusion
to Brahmachari (2011), this resemblance of Based on the results, it can be concluded
action is happened through a biochemical that black rice bran extract with dose of 200
process by facilitating the transport of glucose mg/kg of b.w. was able to lower blood glucose
into the tissues and promotes the synthesis of levels up to 131.33 ± 8.08 mg/dL after 10 days
glycogen as an energy reserve. Mechanisms of of treatment.
flavonoids as a diabetes drug is by stimulating

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