The dynamics of amylolytic enzyme activity in the content of blood and liver homogenate in rats of different age in experimental hypokinesia condition Текст научной статьи по специальности «Фундаментальная медицина»

Dumaeva Zukhrakhon Nasritdinovna, Ph D., Researcher, Ministry of higher and secondary specialized education of Republic of Uzbekistan, Andijan State University E-mail: [email protected] Kadirov Shokir Kadirovich, Doktor of Medical Science Andijan State Medical Institute E-mail: [email protected]

THE DYNAMICS OF AMYLOLYTIC ENZYME ACTIVITY IN THE CONTENT OF BLOOD AND LIVER HOMOGENATE IN RATS OF DIFFERENT AGE IN EXPERIMENTAL HYPOKINESIA CONDITION

Abstract. The purpose of this research is to study the dynamics of amilolytic activity changes in blood and liver homogenate in different age rats under experimental hypokinesis conditions. It was determined that on the 1-day of hypokinesia amylolytic activity in the blood of mature rats rapidly decreased relatively to the control, on the 3-, 10-day it remained in high value and on the 60-day restored at value close to the control, and also on the first day of hypokinesis amylase activity and liver homogenate of mature rats rapidly increased relatively to the control, on the 3-, 10- and 20-day it remained at high value and on the 30-, 60-day it approximated to the value of control. It was found out that on the 1st day of hypokinesia in the old rats, a sharp reduction in amilolytic activity in the blood is significantly higher than that of the control group, on the 3-, 10-day it is significantly higher than that of the control, on the 20-day it reduced again and on the 30-day it changed in restoration and on the 60-day it is at value close to the control. Besides, it was found out that on the first day of hypokinesia in the old rats, the value of amylolytic activity in the liver homogenate decreased in comparison with the control, on the 3-day it approximated to the control, on the 10-day it was higher than that of the control and on the 20-, 30, 60-day it nearly remained at value of control. It was found out that the correlation coefficient between dynamics of changing depending on hypokinesia duration of amylase activity in liver homogenate of young rats is equal to r = 0.29; mature rats: r = 0.32 an old rats: r = 0.58.

Keywords: Hypokinesia, a - amylase activity, blood, liver homogenate.

Introduction 15-462]. Especially hypokinesia of duration has proven to

It is known that fermental homeostasis is one of the have a broad spectrum of complex structures/functional major reactions to various stress factors in the body [1, p. changes in the body [12, p. 991-998; 13, p. 2165-2176; 25-30]. Combined action of stress agents (y-radiation, hy- 14, p. 845-875].

pokinesia, etc.) indicates significant changes in the func- Thus, it is crucial to develop adequate approaches to

tional activity of the hemostasis system [2, p. 286-289]. study and correction of the medical/biochemical mecha-Hypokinesia is one of the most common stress factors in the nisms of dysfunctional changes in the body under hypoki-body, causing specific and nonspecific structure/function- nized conditions [12, p. 991-998; 10, p. 387-389]. Based on al changes in the body [3, p. 40]. Specifically, hypokinesis the information given above, the purpose of this research is has revealed dysfunction in the nervous system, gumoral to study the dynamics of amilolytic activity changes in blood regulator, cardiovascular, respiratory, immune, antioxidant, and liver homogenate in different age rats under experimen-digestive system [4, p. 5-307; 5, p. 3-10; 6, p. 10-224; 7, tal hypokinesis conditions. p. 198-205; 8, p. 3-20; 9, p. 255-267]. In turn, the study Materials and methods

of mechanisms of hypokinesis is one of the topical issues Experimental hypokinesis was performed using standard

in clinical practice [9, p. 255-267]. The research has re- methods in rats [9, p. 255-267; 15, p. 10-270; 16, p. 3; 17, p. vealed that pathogenic morpho-functional changes in the 111-114; 18, p. 117-184; 19, p. 58-65]. cardiovascular system were influenced by hypokinesia [4, p. The preparation of rats homogenate was carried out

25-320]. In addition, the disorder state of the antioxidant using standard methods [18, p. 117-184; 20, p. 351-358] system is influenced by hypokinesis [10, p. 387-389; 11, p. (Fig. 1).

A

B C D

Figure 1. Experimental hypokinesia model (A), preparation of liver homogenate (B), amylolytic activity study (C, D)

ficient correlation of Pirson // [Electronic resource], URL: http://www.machinelearning.ru /wiki/index.php?title... Date of application: 20/01/2019)

n X xy - (Xx) - (X y)_

Effect of a -amylase activity in experiments was determined by means modified by Ugolev A. M., developed by Smithson-Roy [21, p. 345-544; 22, p. 192-196; 23, p. 86-89]. The amylolytic activity was determined by photoconductor method based on the amount of starch dissolved over the given time [24; p. 1228].

Statistical analysis. The outcomes achieved were statistically retreated by means of Excel 2003 (Microsoft Office; USA) and special program packages OriginPro v. 8.5 SR1 (EULA, Northampton, MA 01060-4401, USA). The results were given in the form of M ± m of the results experiments carried out repeatedly n times and indicate M -average arithmetic value and m - standard mistake value. The statistical reliability of the control and experimental group values was calculated on the basis of Student f-scores and was statistically evaluated as reliable at < 0.05 [25, p. 12-351; 26, p. 23-284; 27, p. 15-144; 28, p. 24-459; 29; p.106; 30, p. 54-260]. In the experiments, the likelihood of a link between the dynamics of changes in the amilolytic activity in the structure of the blood and liver homogenates in the hypokinesis environment was discussed in Excel 2003 (Microsoft Office, USA) and OriginPro v. 8.5 SR1 (EULA, Northampton, MA 01060-4401, USA) the Pyron Correlation coefficient was calculated using the following formula with the mathematical statistical program (Coef-

r = -

¿n X x2 - (X x )2 ] [n X y2 - (X y )2 ]

Results and Discussion

Amylolytic enzymes in the blood are one of the most important indicators of the functional activity of the digestive system in terms of clarity in physiological norms/pathological conditions [31, p. 17-25; 32, p. 17-22]. The experiments showed that amilolytic activity of hypokinesia in the bloodstream in young rats increased in 1-, 3-, 10-, 20- and 30-days for 12.1 ± 0.1; 122 ± 1.2; 257 ± 3.7; 24 ± 2.3% and 143 ± 2.6% respectively. In addition to this, in the 60h day of experimental hypokinesia, amylolytic activity in the bloodstream in young rats was restored to the initial state (97 ± 2.7%), which was close to the control group (Tabl. 1).

The subsequent experiments showed that in the liver homogenate of young rats increased in 1-, 3-, 10-, 20- and 30-days of experimental hypokinesia amylolytic activity in-creaded to 16 ± 3; 35 ± 2.1; 1 ± 0.8% respectively with the control. Also, in the 20-, 30-, and 60-day experimental hypokinesia - 22 ± 2.6, respectively; 9 ± 1.2; A decrease of7 ± 1.2% was observed (Tabl 1).

Table 1. - Effects of hypokinesia on amylase activity in the structure of young rats and their liver homogenate (M ± m)

Hypokinesia Blood content (mg/min./g) Liver homogenate (mg/min./g)

duration (day) Control group Experimental group Control group Experimental group

1 2 3 4 5

1 111 ± 0,7 101 ± 0,7 2372±20 2757 ± 679(0,01)

100 12,1 ± 0,9(0,001) 100 116 ± 3(0,01)

3 71,9 ± 2,0 160 ± 1,2(0,001) 2469±41 3338 ± 70(0,001)

100 222 ± 1,0(0,001) 100 135 ± 2,1(0,001)

10 87,0 ± 1,1 311 ± 3,3(0,001) 2736±11 2766 ± 22(0,1)

100 357 ± 3,7(0,001) 100 101 ± 0,8(0,1)

1 2 3 4 5

20 106 ± 1,5 100 131 ± 3,3(0,001) 124 ± 2,3(0,001) 2965± 15 100 2330 ± 60(0,01) 78 ± 2,6(0,001)

30 108,3 ± 2,0 100 155 ± 4,0(0,001) 143 ± 2,6(0,001) 2323± 17 100 2112 ± 26(0.01) 91 ± 1,2(0,01)

60 111,2 ± 3,0 100 108 ± 3,2(0,1) 97 ± 2,7(0,1) 2407± 12 100 2229 ± 27(0,01) 93 ± 1,2(0,01)

Note: The numerators show amylase activeness (absolute value); the denominators show their indicators in percentage; statistic reliability degree of experimental group value (р) versus control groupis in the brackets (In all cases р < 0.05; n = 3-6)

Based on the results of the study, it was noted that the dy- time, in the 1st, 3rd, and 10th days of hypokinesia, an increase

namics of changes in the duration of hypokinesis (1-60 days) in amylolytic activity in the bloodstream was observed with a of amylase activity in the young rats blood and liver homog- linear description in the rats (maximal up to about 3.5 times enates have specific characteristics (Fig. 2. A, B). At the same in control), and 20-, 30-, and 60-days (Fig. 2 A).

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Figure 2. Dynamics of change in amylase activity hypokinesis duration (1-60 days) in the blood (A) and liver homogenat of young rats (B). The ordinate axis shows the percentage of amylase expressed in percentage to 100% controlled. * - relatively to the control p < 0.001, ** - p < 0.01 (n =3-6)

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Also, it was noted that in the young rats, the amilolytic activity in the liver has a dynamics of change due to the hypokinesis duration. In particular, the increase in the amylolytic activity in the liver homogenate within the 1st, 3rd and 3rd days (maximally up to ~16-35% in control), return to the control group for 10 days, decrease to 22 ± 2.6% in the 20-day period and 30- and approximated to control values for 60 days, but lower (9 ± 1.2 and 7 ± 1.2% respectively) in the control (Fig. 2 B).

Also, the coefficient of correlation between the change in the amilolytic activity value of hypokinesis in the liver homogenate in young rats was r = 0.29 (Fig. 3).

Amylolytic activity in the liver or gastrointestinal tract; abscissa axis is the amilolytic activity of the blood (n = 3-6).

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Amylolytic activity in the blood (%)

Figure 3. The coefficient of correlation between the dynamics of change due to the hypokinesis duration of the amilolytic activity in the liver structure of the young rats

In the experimental experimental group, it was determined that amylolytic activity of mature rats was ~8.3 times higher in the blood of mature rats and ~1.7 times less in the liver. The experiments showed that amylolytic enzyme activity in blood of mature rats decreased by 60 ± 1% than the

control on the 1-day, on 3-, 10-day increased by 27 ± 1.4 and 17 ± 0.8%, on 20-day it returned to the control value and on 30-day and 60-day it came close to the control value (respectively - 99 ± 0.7; 101 ± 1%) (Tabl. 2).

Table 2.- The effect of hypokinesia on amylase activity in blood structure and liver homogenate of mature rats (М ± m)

Hypokinesia duration (day) Blood content (mg/min./g) Liver homogenate (mg/min./g)

Control group Experimental group Control group Experimental group

1 784,2 ± 6,2 100 468,8 ± 3(0,001) 60 ± 1,0(0,001) 1401,8 ± 14 100 3233 ± 12(0,001) 230 ± 0,1(0,001)

3 780 ± 6,4 100 992,8 ± 14(0,001) 127 ± 1,4(0,001) 1500 ± 6 100 2214 ± 18(0,001) 148 ± 0,8(0,001)

10 790 ± 7 100 923,8 ± 6(0,001) 117 ± 0,8(0,001) 1519,8 ± 7 100 3240 ± 16(0,001) 213 ± 1,1(0,001)

20 750 ± 6 100 745 ± 7(0,1) 100 ± 0,8(0,1) 1526 ± 8 100 1861 ± 9(0,001) 122 ± 0,6(0,001)

30 730 ± 4 100 722 ± 6(0,1) 99 ± 0,7(0,1) 1604± 10 100 1500 ± 6(0,001) 93 ± 1,0(0,001)

60 730 ± 6 100 738,6 ± 10(0,1) 101 ± 1(0,1) 1670±11 100 1617 ± 7(0,05) 97 ± 0,9(0,05)

Note: The numerators show amylase activeness (absolute value); the denominators show their indicators in percentage; statistic reliability degree of experimental group value (p) versus control group is in the brackets (In all cases p < 0.05; n = 3-5)

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Figure 4. Dynamics of changing depending on hypokinesia duration (1-60 day) of amylase activity in blood

and liver homogenate of mature rats. The ordinate axis shows the percentage of amylase expressed in percentage to 100% in those in control group. * - relatively to the control р < 0.001, ** - р < 0.01 (n = 3-5)

The following experiments showed that on 1-, 3-, 10-and 20-days of experimental hypokinesia amylolytic enzyme activity in liver homogenate of mature rats increased by 130 ±

0.1; 48 ± 0.8; 113 ± 1.1; 22 ± 0.6% respectively to the control, and also on 30- and 60-days it decreased by - 7 ± 7 Ba 3 ± ± 0.9% respectively to the control (Tabl. 2).

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Also, it was found that amylase activity in the blood and liver of mature rats was significantly higher on the first day of hypokinesia than those in control groups, and it was found to be high on 3-, 10- and 20-day and approximated to control value on 30-60* day (Fig. 4 B).

The correlation coefficient between the change in the amylolytic activity value in the liver homogenate of mature rats and the dynamics of change due to the hypokinesis duration is r = 0.32 (Fig. 5).

The experiments showed that the value of amilolytic activity in blood of old rats in the control group is ~7.7 times higher, amilolytic activity in the liver homogenate is ~1.4 times higher in comparison with young rats. In the experiments it was found out that on the 1-day of experimental hypokinesia amylolitic activity in blood of old rats is 61 ± 0.9% lower than the control, and on the 3-, 10-day it increased in -12 ± 1.1 and 11 ± 1.1% in comparison with the control, on the 20-, 30-day it changed up to - 59 ± 1.2; 73 ± 1,1 respectively to those in control group and on the 60-day it approximated to the control value in 96 ± 0.8% (Tabl. 3).

Further experiments showed that on the 1- and 3-days amylolytic enzyme activity in the liver homogenate of old rats decreased from - 70 ± 1.1 up to 95 ± 0.6% respectively to those in control group, on the 10-days it increased in 20 ± 1.7% than those in control group, on the 20-, 30-day it equalized with the control value and on the 60-day it was higher in 5 ± 1.1% (Tabl. 3).

Table 3.- The effect of hypokinesia on amylase activity in blood and liver homogenate of old rats (M ± m)

Hypokinesia Blood content (mg/min./g) Liver homogenate (mg/min./g)

duration (day) Control group Experimental group Control group Experimental group

1 800 ± 19 48,5 ± 7,0(0,001) 3630±45 5214 ± 46(0,001)

100 61 ± 0,9(0,001) 100 70 ± 1,1(0,001)

3 815 ± 17 913,0 ± 10(0,01) 3670±63 3487 ± 19(0,05)

100 112 ± 1,1(0,001) 100 95 ± 0,6(0,001)

10 836 ± 18 928,6 ± 9,0(0,001) 3665±60 4409 ± 60(0,001)

100 111,0 ± 1,1(0,001) 100 120 ± 1,7(0,001)

20 717 ± 16 4292 ± 9,0(0,001) 3710 ± 56 3714 ± 27(0,1)

100 59 ± 1,2(0,001) 100 100 ± 0,8(0,1)

30 720 ± 15 527,0 ± 8,0(0,001) 3620±60 3644 ± 25(0,1)

100 73 ± 1,1(0,001) 100 100 ± 0,7(0,1)

60 710 ± 18 684,2 ± 6,0(0,1) 3567±60 3739 ± 40(0,1)

100 96 ± 0,8(0,1) 100 105 ± 1,1(0,1)

Note: The numerators show amylase activeness (absolute value); the denominators show their indicators in percentage; statistic reliability degree of experimental group value (p) versus control group is in the brackets (In all cases p < 0.05; n = 4)

Based on the achieved results, when the dynamics system of the old rats were analyzed, it was observed that of changes in the duration of the hypokinesis duration hypokinesia had a significant decrease in amilolytic activity (1-60 days) of amylase activity in blood and liver hepatic in the bloodstream of old rats on day 1, significantly high on

At the next stage, the dynamics of changes in the duration of the hypokinesis duration (1-60 days) of amylase activity in blood of the mature rats and liver hepatic system were analyzed (Fig. 4 A, B). In particular, it was observed that hypokinesia had a significant decrease in amilolytic activity in the bloodstream on day 1, significantly high on 3-, 10-day and restored at a controlled rate on 60-day (Fig. 4 A).

Amylolytic activity in the blood (%)

Figure 5. The coefficient of correlation between the dynamics of change due to the hypokinesis duration of the amilolytic activity in the liver structure of mature rats

Amylolytic activity in the liver or gastrointestinal tract; abscissa axis is the amilolytic activity of the blood (n = 3-5)

3-, 10-day and it decreased again on 20-day and restored on 30-day and it is at a controlled rate on 60-day (Fig. 6 A).

Also, it was found that amylase activity in the blood and liver of old rats decreased on the first day in hypokinesia con-

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Figure 6. Dynamics of changing depending on hypokinesia duration (1-60 day) of amylase activity in blood (A) and liver homogenate (B) of old rats. The ordinate axis shows the percentage of amylase expressed in percentage to 100% controlled. The abscissa axis expresses experiment groups (I - control; II - 1-day; III - 3-day; IV - 10-day; V - 20-day; VI - 30-day; VII - 60-day). * - relatively to the control p < 0.001, ** - p < 0.01 (n = 4)

It is determined that the correlation coefficient between dynamics of changing depending on hypokinesia duration of amylase activity liver homogenate of old rats is equal to r = 0.58. (Fig. 7).

125n Correlation coefficient r=0,58

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Figure 7. The correlation coefficient between dynamics of changing depending on hypokinesia duration of amylase activity in liver homogenate of old rats

The ordinate axis shows amylase activity in liver homogenate; The abscissa axis shows amylase activity in blood (n = 4).

It is recorded that significant changes occur in the structure/functional state of the liver in pathological cases in the body [29, p. 8].

Specifically, it is determined that in the initial study of hypokinesis, the activity of the dehydrogenase enzymes in

the liver in the citrate cycle increased, and the activity of liver enzymes in the subsequent stages decreased [33, p. 309-315].

The effect of hypokinesis on the liver and myocardium tissue was determined by the diminished antioxidant system enzymes and glycogen levels (energy deficiency) and the dysfunction of antioxidant system enzymes [17, p. 11-114; 18, p. 17-184].

In general, the effect of hypokinesis on the activity of almost all functional systems in the body is noted [34, p. 4-18; 35, p. 3-40]. In its turn, biological organisms are characterized by various stress-factors, including hypokinesis response, with the function of stress-relieving and stress-limiting systems [36, p. 20-452].

In hypokinesia, the hypothalamic-hypophysic-corticoid system, which is activated by stress is activated and the organism reacts to the condition. The increase in corticosterone concentration in blood determines the specific condition of the rate of hypersus in hypochinized conditions. Also, the influence of endogenous opioids directed against hypokinesis (a -, ft -endorphins, methenephaline, etc.) in the body is activated [19, p. 58-65].

The experimental hypokinesia (1-60 days) may be explained by the difference in the dynamics of amilolytic activity in the blood and liver in the heifoglobin of various age rats, depending on the physiological age and the degree of development of functional systems in the body, as well as the functional activity of compensatory mechanisms.

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Conclusion

Thus, it has been established that amylolytic activity in the blood and liver homogenate of the various age rats has specific characteristics of change due to the hypokinesis duration. Moreover, it was found out that hypokinesia increased in linear classification relatively to the control of amylolytic activity in the blood of young rats on 1-, 3-, 10-days (maximal ~3,5 times than the control), and it restored in close value to the control on 20-, 30- and 60-days. In addition, Also in the young rats, it was noted that the amylolytic activity of the liver has a dynamics of change due to the hypokinesis duration, ie increase in control over the 1st, 3rd, 10th, 20th and 30th days, and 20-, 30-60th day decrease in control.

It was determined that on the 1-day of hypokinesia amylolytic activity in the blood of mature rats rapidly decreased relatively to the control, on the 3-, 10-day it remained in high value and on the 60-day restored at value close to the control, and also on the first day of hypokinesis amylase activity and liver homogenate of mature rats rapidly increased relatively

to the control, on the 3-, 10- and 20-day it remained at high value and on the 30-, 60-day it approximated to the value of control.

It was found out that on the 1st day of hypokinesia in the old rats, a sharp reduction in amilolytic activity in the blood is significantly higher than that of the control group, on the 3-, 10-day it is significantly higher than that of the control, on the 20-day it reduced again and on the 30-day it changed in restoration and on the 60-day it is at value close to the control. Besides, it was found out that on the first day of hypokinesia in the old rats, the value of amylolytic activity in the liver homogenate decreased in comparison with the control, on the 3-day it approximated to the control, on the 10-day it was higher than that of the control and on the 20-, 30, 60-day it nearly remained at value of control.

It was found out that the correlation coefficient between dynamics of changing depending on hypokinesia duration of amylase activity in liver homogenate of young rats is equal to r = 0.29; mature rats - r = 0.32 an old rats - r = 0.58.

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