EVALUATION
OF ANTIOXIDANT PROPERTIES OF NEW FERMENTED MILK PRODUCT BASED ON THE MARE'S
MILK IN PATIENTS WITH NEW-ONSET PULMONARY TUBERCULOSIS
Yu. A.
Sinyavsky Kazakh Academy of Nutrition, Kazakh National Medical University named
after S.D. Asfendiyarov, A. S. Targautov, Kazakh Academy of Nutrition, V. G.
Vyskubova Kazakh Academy of Nutrition, M. S. Puchkova Kazakh Academy of
Nutrition, Yu. N. Bespalova Kazakh Academy of Nutrition, D. A. Sukenova Kazakh
National Medical University named after S.D. Asfendiyarov
Abstract—A new multi-functional product based on the mare's
milk for the alimentary support of patients with pulmonary tuberculosis is
developed. Independent clinical trials have shown the efficacy of fermented
milk product which was in a more rapid elimination of intoxication syndrome,
leukogram normalization, increase of immune status.
Key words: antioxidants, mare's milk,
tuberculosis.
Tuberculosis remains one of
the most urgent problems of the modern world community and today is recognized
by the World Health Organization (WHO) as the disease requiring the development
and implementation of active and immediate actions to prevent the spreading in
the world [1]-[4].
Problem
of the relationship of tuberculosis and nutrition taking into account the main
trends in the development of world science and international experience is
unquestionable and addresses the important nutritional disorders, determines
the severity, course, low effectiveness of specific therapy as well as
vaccination [5].
In
order to improve the efficacy of specific chemotherapy it is necessary to use
wider the methods of nutritional (alimentary) support which allow for
significant increasing of the effectiveness of etiological and pathogenetic
therapy as well as to reduce the risk of side effects largely determining the
ultimate result of treatment. Here the greatest prospect is a reasoned
application of functional foods and dietary supplements, correcting basic
systems of organism adaptation (hematopoietic, immune, antioxidant,
monooxygenase, normalizing the microbiocenosis function of intestines, etc.).
Taking
into account the high nutritive and biological value of mare's milk as well as
available information regarding the use of fermented milk product based on the
mare's milk-kumiss some practical interest was directed to the development of a
new multi-functional product based on the mare's milk and evaluation of its
clinical effectiveness in tuberculosis practice.
On the basis of pulmonary therapeutic
department No. 3 of National Centre for Tuberculosis of the Ministry of Health
of the Republic of Kazakhstan the clinical efficacy of a new fermented product based
on the mare's milk "Tulpar-2" was evaluated in 25 patients with
new-onset pulmonary tuberculosis who obtained fermented milk product during the
second week after beginning of chemotherapy (isoniazid + rifampicin +
pyrazinamide + ethambutol). Patients of the control group (25 persons) received
the standard chemotherapy in mode of the 1 category; against the overall drug
therapy the patients received 400 ml of fermented milk (kefir) in two doses of
200 ml during 2 months. Groups were comparable on sex and age composition and
clinical diagnosis.
The
intensity of lipid peroxidation processes and enzymatic activity level of
antioxidant protection (catalase and superoxide dismutase) evaluated in red
blood cells and blood plasma before and after taking a fermented product
"Tulpar-2". Catalase activity was spectrophotometrically measured at
a wavelength of 240 nm (370Ñ) from the
rate of disappearance of H2O2 using test kits of “Sigma”
(Germany). Superoxide dismutase activity (SOD) was also spectrophotometrically
determined at 540 nm (370Ñ) from the
rate of recovery of nitro blue tetrazolium (NBT) to blue diformazan in xanthine
oxidase reaction [6], [7].
MDA level was determined using the
test kits from “Sigma” (Germany) according to Yu. A. Vladimirov and A. I. Archakov
[8], [9].
Diene conjugates concentration was
determined according to the procedure of V.B. Gavrilova and M. I. Meshkorudnaya
[10].
All the digital material was subjected to mathematical processing with
methods of analysis of variance, in some cases non-parametric statistics tests
were used [11], [12].
In
assessing of lipid peroxidation (LPO) processes under the primary and secondary
products data the results characterizing the modifying effect of alimentary
support of pathogenetic therapy with a specialized product "Tulpar-2"
(Table I) were obtained. Initially LPO processes in patients with tuberculosis
were characterized by a high level of primary and secondary products (MDA and
diene conjugate of plasma). Thus, comparing with values of apparently healthy
individuals the level of MDA in plasma of patients was 1.9 times higher, or by
91.7%. Diene conjugates in tuberculosis patients compared with healthy
individuals, were also 1.7 times higher, or by 69.9%. At that the changes of
values of malondialdehyde and diene conjugates in tuberculosis patients
compared with healthy people were accurate.
TABLE I
Changes of
some blood biochemical parameters in patients with tuberculosis before and
after receiving the specialized product "Tulpar-2", compared with
healthy individuals (M ± m)
|
Indicators |
Healthy
individuals |
Before receiving |
After receiving |
|
TC, mmol / l |
5.6±0.2 |
6.7±0.4 |
5.8±0.4 |
|
Triglycerides, mol /l |
1.3±0.2 |
1.5±0.2 |
1.4±0.3 |
|
Low-density lipoproteins cholesterol, mol/ l |
3.0±0.4 |
5.2±0.4õ |
4.0±0.5 |
|
Diene
conjugates, µmole/l |
15.6±1.2 |
26.5±2.8õ |
20.4±0.3 |
|
MDA, μmole/l |
2.40±0.3 |
4.6±0.5õ |
3.6±0.4 |
|
GP, activity unit/ ml of blood |
3.90±0.3 |
2.8±0.5 |
3.5±0.2 |
|
SOD, activity unit/ml of blood |
256±20.6 |
202.3±11.1 |
270.3±13.0 |
|
Catalase, activity unit / ml of blood |
43.5±3.6 |
64.3±2.7õ |
55.2±3.3 |
|
õ- the
results are statistically significant in relation to the results obtained in
healthy individuals. |
|||
During
treatment in two months the patients taking a specialized product have
improvement towards normalization of the aforementioned indicators. Thus, the
level of MDA decreased from 4.6 ± 0.5 to 3.6 ± 0.4 mmol / ml, or by 20.6%,
diene conjugates - from 26.5 ± 2.8 relative units to 20.4 ± 0.3 relative units,
or by 23%, while that the changes were not accurate, but all the same there was
improvement in almost all the examined patients, indicating a favorable effect
of the product to the antioxidant status.
In
parallel with the high level of LPO products was an increase in the activity of
catalase (CT) in the blood plasma of patients with tuberculosis by 47.8%,
compared with healthy individuals. Activity of glutathione peroxidase (GP) and
superoxide dismutase (SOD) in erythrocytes of patients with tuberculosis by
contrast decreased by 25.6 and 21%, respectively, compared with healthy
individuals. Increase in catalase activity was probably due to the excessive
accumulation of hydrogen peroxide as a result of the active oxidation of the
lipid phase of membranes, and decrease of activity of SOD and GP could be due
to a decrease of the level of selenium, vitamin E, glutathione and other
antioxidants in the blood and tissues of patients which directly meant the
reasonable correction with antioxidants of the diet of patients with
tuberculosis.
After
two months of taking the specialized product "Tulpar-2" on a
background of drug therapy was a tendency to decrease in catalase activity by
14.2% in patients of the main group, the activity of SOD and GP thus increased
by 33.6 and 25.0%, respectively.
Also
it should be noted that, with respect to the healthy individuals, the patients
with tuberculosis have an increase of level of total cholesterol,
triglycerides, low density lipoprotein cholesterol in blood by 17.9, 15.4 and
73.0 %, respectively. On the background of taking the fermented milk product
"Tulpar-2" the patients showed a decrease of level of total
cholesterol and low density lipoprotein cholesterol by 13.4 and 19.2 %,
respectively. Despite the fact that these figures have changed insignificantly
and have not reached the values in healthy persons, however, improvement
pointed to the antiatherogenic effect and antioxidant properties of this
alimentary remedy.
Assessing
the level of secondary LPO products (diene conjugates and malondialdehyde) in
the blood of patients of experimental group compared to the control one, has
revealed their significant reduction after the course of diet therapy with
fermented milk product "Tulpar-2" (Table II). The control group also
outlined the tendency to reduce the level of secondary and end LPO products,
but it was not accurate.
TABLE II
Changes in
measures of LPO- antioxidant defense in blood plasma of control and
experimental groups of patients with pulmonary tuberculosis (M ± m)
|
Indices |
before treatment |
after treatment |
||
|
Main group n=21 |
Control group n=21 |
Main group n=21 |
Control group n=21 |
|
|
MDA,
μmole/ml |
4.6±0.5 |
4.3±0.5 |
3.6±0.4 |
4.0+0.4 |
|
DC, μmole/l |
26.5±2.8 |
25.6±1.7 |
20.4±0.3 |
25.0±1.5 |
|
HP activity
unit / ml of
blood |
2.8±0.5 |
3.0+2.7 |
3.5±0.2 |
3.2±0.2 |
|
CT activity unit / ml of blood |
64.3±2.7 |
68.7+2.2 |
55.2±3.3 |
65.1+0.46 |
|
SOD activity
unit / ml of
blood |
202.3±11.1 |
208.3+10.2 |
270.3±13.0 |
227.8+9.7 |
|
*
- differences are statistically significant in relation to the data before
receiving the "Tulpar-2" specialized product. |
||||
After
taking the product the level of catalase reduced, activity of antioxidant
enzymes (SOD and GP) increased. In the control group, the figures have changed,
but the percentage of their shifts was much lower than in the experimental
group.
In
the control group, the parameters of individual enzymes have slightly changed
in a significant part of the patients, which determined insignificant decrease
of average group values.
Based
on the obtained results it is found that the use of specialized product
"Tulpar-2" in the complex treatment of patients with new-onset
pulmonary tuberculosis contributed to the normalization of metabolic disorders,
leading to stabilization of processes of free radicals oxidation- antioxidant
defense of blood followed by providing more pronounced immunomodulatory effect,
as compared to the patients of control group.
Thus,
according to the test of fermented milk product "Tulpar-2" as a
pathogenic drug, we observed the improvement of the treatment effectiveness of
patients with new-onset pulmonary tuberculosis, which was expressed by more
rapidly elimination of intoxication syndrome, normalization of leukogram,
increase of immune status and normalization of processes of LPO-antioxidant
defense.
The
resulting changes have been associated not only with a favorable influence of
mare's milk on the basic parameters of antioxidant defense and immunity, but
also with the positive influence of antioxidant vitamins, selenium, PUFAs, soy
isoflavones, bioflavonoids, pectin, inulin, macro- and micronutrients to
immunity, antioxidant status, organism deoxidation system, state of intestines
microbiocenosis and metabolic processes in general. This product can be
recommended for routine use in TB practice.
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