Medicinus - Dexa Medica

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research original article
glucose were purchased from Merck.
Animals Experiment
Wistar strain rats (Rattus norvegicus) (male, 8 weeks
old, 200-220 g) were housed in individual cages in
a room with controlled temperature and light (12h
light:12-h dark cycle; lighting between 08:00 and
20:00 h) and had free access to a standard diet and
water. The animal study was conducted according
to the regulation proposed by AAALAC (Association
for Assessment and Accreditation of Laboratory Animal
Care). The rats were aclimated for a week before
treatment.
Experimental Design
Rats were allocated into 3 groups. The first group
(n=7) was control group that was maintained on a
standard diet. The second (n=7) and the third group
(n=7) were maintained on a high-glucose diet for
two weeks to make them insulin resistant with fructose
15% and glucose 10%. The third group was later
on treated with DLBS3233 at 9 mg/kg body weight
for another two weeks. At the end of treatment period,
blood samples were collected from tail veins of
rats to measure the random glucose level. The rats
of each group were then fasted overnight, fed and
2 h later their blood samples were taken to measure
the postprandial glucose. Furthermore, animals
were fasted overnight again and blood sample was
later collected by cardiac puncture which then was
centrifuged at 8500 g for 25 min. The serum was
separated out to measure the level of fasting glucose,
insulin, triglycerides, total cholesterol, Low
Density Lipoprotein (LDL), and High Density Lipoprotein
(HDL). Mercodia Ultrasensitive Rat Insulin
ELISA enzyme immunoassay (Uppsala, Sweden) and
microplate reader (Bio-Rad) were used to measure
the insulin level. Other tests were carried out using
Semi Automated Clinical Chemistry Analyzer Microlab
300 (Vital Scientific, Netherlands). In addition,
group glucose (mg/dl) Insulin
Random Postprandial Fasting
(mg/dl)
MEDICINUS 24(1), January 2011
Homeostasis Model Assessment (HOMA) was calculated
by multiplying the glucose (mmol/L) with the
insulin concentration, followed by dividing it with a
factor of 22.5.7 Data was analyzed using ANOVA test
and significance level was measured using posthoc
Bonnferoni from Biostat software.
Data statistics
Results were expressed as mean ± standard deviation.
To confirm the normal distribution, the data
were analyzed with normality test and subjected to
statistical analysis using the ANOVA test. To measure
statistical tests, posthoc Bonnferoni from Biostat
software was used.
RESULTS
Wistar strain rats were treated with normal saline per
oral as the control group in this experiment. Fructose
15% and glucose 10% were given to the animals
to stimulate insulin resistance condition, while
DLBS3233, 9 mg/kg of body weight, was given as a
treatment on top of fructose-glucose intake.
Administration of fructose and glucose increase
the concentration of blood glucose level to 41.33%
for random glucose level, 49.02% for postprandial
glucose, and 60.22% for fasting glucose. Total cholesterol
in blood was increased by 59.59%, similar
profile was also seen in insulin level which was increased
to 220.38% (Fig. 1), LDL concentration in
blood to 31.70% and triglycerides to 61.39%. Moreover,
the HDL level decreased up to 17.15% (Table 1).
Treatment with DLBS3233 for 2 (two) weeks decrease
the concentration of random glucose, postprandial
glucose, fasting glucose, insulin, total cholesterol,
LDL, and triglycerides by 29.64%, 30.62%,
31.41%, 65.01%, 33.78%, 21.36%, 30.81%, respectively,
compared to positive control groups which
have been treated with fructose 15% (Table 1 and
Fig. 1). On the other hand, the level of HDL was increased
by 18.20% (Table 1) .
Total
Cholesterol
(mg/dl)
LDL
(mg/dl)
HDL
(mg/dl)
Triglycerides
(mg/dl)
Control 84.23 ± 5.5 76.87 ± 7.73 69.83 ± 4.52 64.63 ± 3.68 168.29 ± 10.84 156.56 ± 3.76 28.04 ± 1.33 59.97 ± 6.98
Insulinresistant
Table 1. Effect of DLBS3233 on the level of glucose, insulin, total cholesterol, LDL, HDL and triglycerides
119.04 ± 7.54 114.55 ± 8.36 111.88 ± 7.65 207.06 ± 13.63 268.57 ± 11.86 206.19 ± 6.38 23.23 ± 1.33 97.11 ± 5.75
Treated 83.76 ± 8.95 79.48 ± 9.93 76.74 ± 8.63 72.45 ± 11.49 177.86 ± 11.1 162.14 ± 6.67 28.40 ± 1.34 67.19 ± 5.35