Rat Blood Lipid Improvement by Using Soy-Based Yogurt on Probiotic Drink

Lovita Adriani1, Jajang Gumilar1, Hendranoto.A.W Lengkey1, Novi Mayasari1

1
Faculty of Animal Husbandry, Universitas Jl. Raya Bandung Sumedang KM 21, Jatinangor,
West Java, 45363, Indonesia
Corresponding Email.j.gumilar@unpad.ac.id

Abstract
Background : Probiotics are a microorganism that can modify the microflora in the body, it can
replace the pathogenic microbe with non-pathogenic microbe for the body in the human’s
intestine and has some functions to improve blood lipid, to repair immune system and to act as
antioxidant.
Method : Cow milk and soy milk were pasteurized for 15 minutes at 73C, then cooled to
43C for incubation, and added with 2% culture starters, a mixture of Lactobacillus bulgaricus :
Lactobacillus acidophilus : Streptococcus thermophilus = 1 : 1 : 1, until the desired texture with a
pH between 4.0 to 4.5. Four treatments, consisted of the ratio of cow milk and soy milk: T0=
100% cow milk, T2 = 80% cow milk + 20% soy milk, and T3 = 70% cow milk + 30% soy milk;
with 6 replications. Blood samples of rat were collected for blood lipid (cholesterol, triglyceride,
high density lipoprotein (HDL) and low density lipoprotein (LDL)) analysis.
Result : Based on ANOVA test, there were no significant differences on level of cholesterol,
triglyceride, HDL and LDL in rat blood among treatments. However, the use of soy milk in soy-
yogurt reduced plasma cholesterol and triglyceride. It seems that fermentation processes of soy
milk by probiotic activate β-glucosidase. This enzyme will hydrolyse isoflavone to aglycones.
Aglicones prevent chronic diseases like coronary heart disease.
Conclusion : In conclusion, intake of soy-yogurt improved rat blood cholesterol and triglyceride
even though there were no significant differences. Moreover, HDL and LDL levels were similar
among treatments.

Keyword : blood lipid, soy milk, cow milk and probiotic

Introduction
Probiotic is living microorganism which are often used as an additional feed and has a
function to improve health by balancing the microflora in the intestine. The benefit of probiotic
consuming can be able to inhibit the population of pathogenic bacteria, to provide enzymes
which have a function in digestion, and to stimulate intestinal peristaltic activity 1. One product
which contains probiotic and can be given to a rat is yogurt. Yogurt is one of the fermentation
products which is made up of milk as the base material and is fermented by one or several
microorganisms. However, in this research, we will use cow milk and soy milk as the base
material of yogurt because a prior research has proved that soy milk has a high benefit for the
body.
The application of probiotic in the yogurt can offer a positive impact on health, because
probiotic in the yogurt can maintain the blood lipid level. Then, probiotic also has a function to
reduce body weight, adipose fat size, serum leptin, and blood glucose level 4. Blood lipid consists
of cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), and triglyceride
(TG). The abnormal level of blood lipid may cause several health problems like heart diseases
and obesity. The consumption of yogurt can be one of the solutions to prevent the abnormal level
of blood lipid because several studies reported that probiotics significantly reduce TC and LDL
levels, and improve the HDL concentrations 2. Besides that, probiotic can result from statin that
is inhibitor compound 3-hidroxy-3-methyl-glutaryl-CoA reductase (HMG-CoA reductase) which
plays a role to control cholesterol biosynthesis, lowering LDL, VLDL, and blood triglyceride
level3.
Research using cow's milk-based yogurt has been carried out on blood lipids. However,
there have not been many studies on the effect of yogurt based on cow's milk and soy milk on
blood lipid. Therefore, this study is critical to prove which treatment affects lipid improvement.

Material and Method

Animal : The research used 30 female rats with 4 weeks of maintenance. The sample of blood
was taken at the end of the research, with each treatment was repeated 10 times.
Research Material: Cow milk and Soy milk were pasteurized for 15 minutes in the temperature
73oC, then it was cooled to temperature 43oC. Afterward, it was added with 2% culture starters.
The mixture comprises Lactobacillus bulgaricus, Lactobacillus acidophilus, and Streptococcus
thermophiles with comparison 1:1:1 until the desired texture with pH between 4.0-4.5.
Research Method : This research used Completely Randomized Design (CRD) with 3
treatments and 6 replications .
T0 = Control

T1 = Yogurt with the base material of 100% cow milk

T2 = Yogurt with the base material of 80% cow milk + 20% soy milk
The method, that was used in administering yogurt balance, followed the method 5 with a dose of
1.25% of body weight. Cholesterol and triglycerides were analyzed using Biolabo product.
CHOD-PAP (Cholesterol Oxidase Phenylperoxidase Amino Phenozonphenol) method was used
to analyze the cholesterol.

Result
The results from this research is shown in Table 1 below :
Table 1. The Level of HDL, LDL, TC and Triglyceride in Rat Blood
No. Treatment T0 T1 T2
…………………………………….…………..Mg/dL………………………………………….
1 High-Density Lipopoprotein 16.42±3.80 15.33±3.92 18.42±2.05
(HDL)
2 Total Cholesterol (TC) 37.76±2.63 30.90±5.94 32.37±6.05
3 Low-Density Lipopoprotein (LDL) 14.06±4.01 17.74±3.43 13.94±2.64
4 Triglyceride 31.021±5.26 24.62±13.64 22.7±7.05
Note : T0 (Control)
T1 (100% Cow’s Milk)
T2 (80% Cow Milk + 20% Soy Milk)

Based on the ANOVA test is known that all of the treatments in the Tabel 1 do not have a
significant result (P>0.05). However, T2 (80% cow milk + 20% soy milk) tends to improve the
rat blood level in the form of increase HDL and decrease LDL and Triglyceride. Figure 1 is
shown to clarify data from Table 1.

mg/dL
40
35
30
25
20
15
10
5
0
HDL TC LDL Triglyceride

T0 T1 T2
Figure 1. Illustration of Level of HDL,TC,LDL, and Triglyceride in Each Treatment
Discussion
High Density Lipoprotein (HDL) : The average result of HDL in the rat blood is 16.42-18.43
mg/dL. Based on the ANOVA test, the supplementation of soy-yogurt does not give a significant
result (P>0.05) on HDL level compared with control. Althought the result was not significantly
different, T2 tends to improve the level of HDL in rat blood. HDL is the smallest lipoprotein
compound that is produced in the liver and small intestine and has the highest density because it
contains more protein than cholesterol. The content is the most direct apolipoprotein (Apo A-I
and Apo A-11). HDL particle rises because of circulating through the bloodstream and
incorporating more cholesterol and phospholipid molecules from cell , for example, by
interaction with the transporter ABCG1 and Phospholipids Transport Protein (PLTP) 6. However,
in this research there was no increase of HDL in all of the treatments, but there is a tendency of
increasing HDL level in T2.
Total Cholesterol (TC) : The average result of cholesterol is 37.76-32.37 mg/dL, and based on
the ANOVA test, all of the treatment does not give a significant result (P>0.05) on the
cholesterol level in rat blood. However, the level of blood cholesterol is still in the normal range
of 10-54 mg/dL12, and T1 (100% cow’s milk) tends to improve cholesterol level in the rat blood.
The giving of probiotic has not to functioning optimally because it is suspected the inhibition of
nutrient absorption by increasing production of mucus from goblet cell. S.thermophilus and
L.bulgaricus are an ordinary probiotic starter that has a low resistance to acid pH and bile salts 7.
They cannot’t survive when passing through the gastrointestinal tract. This is possible why these
treatments do not have a significant result8. However, the presence of L.acidophilus can give a
positive effect in improving blood cholesterol due to its high resistance to acid8. The mechanism
for the cholesterol-reducing effect of this bacteria includes the inhibition of cholesterol
absorption from the small intestine through binding of cholesterol and bile acids by bacteria
cells, and the assimilation of cholesterol by lactic acid bacteria9.
Low Density Lipoprotein (LDL) : Research result shows the average of LDL level in rat blood
is 14.06-13.94 mg/dL. The administration of probiotic in soy-yogurt doesn’t has a significant
result (P>0.05) on decrease LDL level in rat. However, the LDL level in all of the treatments are
still in the normal range of 7-27,2 mg/dL10, and T2 (80% cow’s milk and 20% soy milk) tends to
decrease the LDL Level. The decrease of LDL level in T2 is caused by the combination of
Cow’s milk and Soy milk. The existence of Lactic Acid Bacteria (LAB) and active substances
contained in the fermented soybean milk can improve the level of LDL in the rat blood. Even
though fermented soybean milk consists of an oligosaccharide, it can’t be used by the LAB as
energy. This is provided by the fermented cow milk as it contains lactoses that can be utilized as
an energy source for LAB5. The fermentation process (in T2) can active isoflavone and alter it
into aglycone by a β-glucosidase enzyme. Aglycone in genistein and daidzein of soybean can
increase bile acid excretion and regulate LDL receptor activity 5,11.
Triglyceride : The average of triglyceride in rat blood is 22.7-31.02 mg/dl. Based on the
ANOVA test, the administration of probiotic in soy-based yogurt does not give a significant
result (P>0.05) on triglyceride level on rat blood. However, the level of triglyceride is still in the
normal range (< 150 mg/dl)11 and if compared to control, the level of triglyceride in T2 tends to
decrease, reflected by lower value of it than T0 (control. The utilization of fermented soybean
milk in the treatments (T2) enables to decrease blood triglyceride level in rat because Soybean
contains isoflavones compound that has a role in activating Peroxisome Proliferator-activated
Receptor-α (PPAR-α). PPAR-α can be able to decrease gene activity in producing triglycerides.
This PPAR- α also rise lipoprotein lipase activity which has a role as lipolysis triglyceride in
chylomicron and VLDL13. Moreover, the content of flavonoid in soybean can inhibit the early
stage of the triglyceride synthesis reaction. Flavonoid will release one of H atom from hydroxyl
acetone phosphate group thats has an ability to bind free radical. Therefore, the activation energy
is reduced resulting in the limitation of the triglyceride oxidation14.
Conclusion
From this research can be concluded if the administration of probiotic in soy-yogurt does not
give a significant result on Rat-blood lipid. However, it can able to maintain the rat-blood lipid
still in the normal range.
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