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Білик О.В. Фармакокорекція препаратом кверцетин впливу гіпоксії та гіпертермії на мінеральний склад кісткової тканини
13.08.2014, 19:30

Резюме
Білик О.В. Фармакокорекція препаратом кверцетин впливу гіпоксії та гіпертермії на мінеральний склад кісткової тканини.
В змодельованому досліді на білих щурах-самцях вивчали фармакотерапевтичну активність кверцетину при дії гіпоксії на фоні гіпертермії на мінеральний склад кісткової тканини.
Ключові слова: гіпоксія, гіпертермія, кверцетин, кісткова тканина.

Резюме
Билык О.В. Фармакокорекция препаратом кверцетин действия гипоксии и гипертермии на минеральный состав костной ткани.
В смоделированном опыте на белых крысах-самцах изучали фармакотерапевтическую активность кверцетина при действии гипоксии на фоне гипертермии на минеральый состав костной ткани.
Ключевые слова: гипоксия, гипертермия, кверцетин, костная ткань.

Summary
Bilyk O.V. Pharmacocorrection preparation quercetin action of hypoxia and hyperthermia on bone mineral composition.
In modeled experiment to study on white male rats pharmacotherapy action of quercetin under the action of hypoxia on the background of hyperthermia on the composition of bone minerals.
Key word: hypoxia, hyperthermia, quercetin, bone tissue.

Рецензент: д.мед.н., проф. В.І. Лузін

УДК 615.356:57.043:611.018.4

ДЗ «Луганський державний медичний університет»

ГЗ «Луганский государственный медицинский университет»

State Establishment "Lugansk State Medical University"

oksana_bilik@mail.ru

Introduction. The study of disorders of bone tissue condition at hypoxia syndrome on the background of hyperthermia has either general biological or practical importance for human health. The tendency to wide distribution of bone system diseases among minors of Donetsk coal field is stipulated by the combination of such unfavorable conditions as low contents of oxygen in air at increased humidity (more than 80%) and temperature of surrounding air(above 300⁰C). At this the disturbance of metabolic processes in loco-motor system, accompanied by the disorder of microarchitechtonics and strength of bones, decrease of bone mass and increase of fracture risk [1,2], is very important.

         Despite the interest of investigators to this problem, only fragmentary scientific reports about hypoxia [3, 4] and hyperthermia [5, 6, 7] effect on bone tissue are observed in modern literature. Simultaneous influence of hypoxia and hyperthermia on bone tissue is not practically studied. And namely the latter conditions the actuality of the study of morphological physiological changes of bone structures under the influence of these factors.

         The aim of the present work was to investigate a simultaneous effect of hypoxia and hyperthermia on rats’ skeleton bones of reproductive period and old ones; and quercetyn application for correction of these factors.

         The connection of thesis subject with scientific programs, plans and themes. The work was performed in accordance with a subject plan of scientific researches of SE” Lugansk state medical university” within scientific research themes of the chair of medical biology “ The regulation of structural tissue homeostasis that is renewed and correction of its changes in conditions of exogenic and endogenic factors influence” (state registration №0199U001828)  and the chair of normal human anatomy “Bone system morphogenesis peculiarities at acute and chronic influence of some environmental factors on the organism” (state registration № C.H.07.02.0001.86), “The peculiarities of growth, structure and shape formation of skeletal bones under the influence of some external and endogen factors at different physiological body conditions” (state registration №0199U001811).

Material and methods of research. The experiment was carried out on 144 white male rats of pubertal age (11 months with body weight-260g) and old ones (24 months with body weight-320g). The animals were separated due to the scheme of the trial (table 1)

Animals Separation

Series

Animals characteristics

Group

Age parameter

Amount  of animals

Time of experiment (days)

10

30

90

I

control

1

Adult of pubertal age

18

6

6

6

2

old

18

6

6

6

II

+ quercetyn

1

Adult of pubertal age

18

6

6

6

2

old

18

6

6

6

III

Hypoxia on the background of hyperthermia

1

Adult of pubertal age

18

6

6

6

2

old

18

6

6

6

IV

Hypoxia on the background of hyperthermia +quercetyn

1

Adult of pubertal age

18

6

6

6

2

old

18

6

6

6

Total

144

48

48

48

 

Preparation dose was calculated due to [8], 100mg/kg of living mass as boluses were given to each animal every day at 7.00 in the morning. Mine condition modeling was created in specially constructed thermo-chamber, in which air temperature was 42-44C and oxygen contents was 6% which was reached by the gas-like nitrogen supply with speed-1,25 l/min during 15 minutes (device “Poisk-2”), air humidity was supported at the level about 80% (psychometer “Asmana”).

         On the completion of trials (10, 30, 90) the animals were taken out of the experiment by decapitation under ether narcosis and skeletation was performed. Investigations were carried out taking into account the demands of “European convention about the protection of vertebrates, used for experiments or other scientific aims” and due to the law of Supreme Council of Ukraine “About animal protection from cruel treatment” [9,10].

         The chemical study of bones included the detection of the contents of mineral and organic substances and water. These indexes were determined through weight method based on the difference between initial and dried out mass in  drying drum at 105⁰C up to constant weight. The water content was determined   on the difference of humid and dried bone weight. Ashalation was performed at 450-500⁰C during 12 hours in muffle drum. The amount of organic and inorganic substances was determined on the difference of dry bone mass and ash. The ash was ground in the mill and kept in hermetic micro test-tubes. For detection of macro elementary contents 5,0 mg of ash was dissolved in 2 ml of chemically pure 0,1 of hydrochloric acid (HCl), brought to 25ml with twice distilled water and spectrometrically determined the contents of calcium, potassium and natrium [11].

         Strength index of studied bones (SIB) was calculated on the formula [12] for integration detection of bone growth parameters, microstructure of skeleton mineral balance:                   where L-is bone length, m- initial mass.

         Statistic processing of obtained data was performed with the help of programme support on PC using mono- or multifactor dispersion analysis (package of licensed “Microsoft Excel”, “Stadia. 6.1/prof” and “Statistica” [13, 14]. We estimated average meanings, their errors, correlation ratio, Student and Fisher criterion and authenticity of statistical indexes.

         Research results and their discussion

         The analysis of obtained data of simultaneous hypoxia and hyperthermia on experimental adult animals of reproductive period (series III, 1) showed that the largest changes were observed on the 30th day of the trial, in particular, there was a considerable decrease relating to the control, concentration of hydrophilic elements in all studied bones. Thus, the amount of potassium in humeral bone decreased    by 16,3%, shinbone – by 24,5%, pelvic – by 44,9 %, third back vertebrae – by 29,3% (p<0,05). The amount of natrium reduced in humerus by 44,6%,      shinbone – 33,5%, pelvic – 32,8%, third back vertebrae – 45,9% (p<0,05).         The water amount also diminished within the limits from 5,1% in pelvic bones to 7,1% in humeral ones (p<0,05).

         On the 90th day, going by the findings of the research, the amount of water continued to decrease in all bones comparing to the control. The amount of potassium and natrium remained lower than a corresponding control one.

         Analyzing the obtained data of old animals (series III, 2) it was revealed that on the 10th day of the study the water contents in all bones reduced while in the shinbone this decrease was probable relatively to the chemical indexes indicated the least water content in all examined bones during the entire experiment. Along with this the amount of hydrophilic element was also decreased- potassium and natrium, comparing to the 10th day of the trial, as well as in comparison with the control.

         Thus, unfavorable factors, namely, hypoxia on the background of hyperthermia, negatively influence the mineral content of bone tissue.

         The application of bioflavonoid quercetyn in dose of 100mg/kg of living mass in animals of pubertal age (series IV, 1) contributed to the strengthening of all surveyed bones.

         Through chemical investigations it was established that the application quercetyn on the 30th day contributed to a significant increase of water amount in all studied bones comparing to the effect of hypoxia and hyperthermia, as well as with control group. On the 90th day these indexes remained higher in comparison with the effect of hypoxia and hyperthermia, but in most bones they were significantly lower than a corresponding control. The amount of potassium and natrium on the 30th day of the experiment increased in all studied bones, comparing to the influence of hypoxia and hyperthermia and reached a corresponding control. On the 90th day the amount of potassium and natrium remained higher than corresponding indexes under the influence of hypoxia and hyperthermia and was on the level of control ones.

         Characterizing the obtained data of old animals (series IV, 2) we can ascertain that mineral saturation of all examined bones was lower in comparison with hypoxia and hyperthermia effect during the experiment. While comparing to the control on the 30th day the amount of mineral substances in most bones exceeds the corresponding indexes and was considerably lower than control index on the 90th day.

         The water amount on the 30th day in all studied bones was lower than hypoxia and hyperthermia effect. On the 90th day the amount water in shinbone and pelvic bones diminished by 5,4% and in back vertebrae by 5,1% (p<0, 05). These indexes in all bones were considerably lower than a corresponding control. The amount of potassium and natrium in all bones varied the same way as at the effect of hypoxia and hyperthermia and was considerably lower than a corresponding control

Conclusions

         Thus, the given data of research testify that the probability of changes in bone tissue depends on the term of simultaneous influence of hypoxia and hyperthermia and the age of animals. The study of quercetyn effect as an antioxidant revealed that its application contributed to the stabilization of examined indexes almost up to the control level.

 

Література

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  2. Herbert G. Oxygen the problem of hypoxia / G. Herbert, Т. Noll // Dioxim. Sos.Trans.1987. – № 3. – P. 363-365.
  3. Ляпков Б.Г. Тканевая гипоксия: клиника – биохимические аспекты / Б.Г. Ляпков, Е.Н. Ткачук // Вопросы мед.химии. – 1995. – № 2. – С. 2-7.
  4. Савченкова Л.В. Биохимические основы патогинеза гипоксического синдрома / Л.В. Савченкова // Укр. мед. Альманах. – 1998. – № 1. – С. 90-99.
  5. Козлов Н.Б. Гипертермия: биологические основы патогенеза, профилактика лечения / Н.Б. Козлов. – Воронеж, 1990. – 187 с.
  6. Особенности формообразования костей осевого и добавочного скелета в условиях хронической гипертермии на разных этапах постнатального онтогенеза / С.М. Смоленчук, Н.В. Корявый, М.Г. Грищук, А.М. Крупник, К.В. Бахилов // Український морфологічний альманах. – 2008. – т. 6, № 2. – С. 196-205.
  7. Лузин В.И. Особенности роста костей скелета белах крыс, подвергшихся воздействию экстримальной хронической гипертермии в сочетании с физической нагрузкой и возможным корректором инозином / В.И. Лузин, С.М. Смоленчук. // Український морфологічний альманах – 2008. – Т. 6, № 2. – С. 52-59.
  8. Расчет основных фармакокинетических параметров на программируемых микрокалькуляторах / Под ред. В.Д. Лукъянчука. – Луганск: РИО, 1990. – 38 с.
  9. European convention for protection of vertebrе animals used for experimental and other scientific purpose. Council of Europe. – (18.03.86). – Strasburg, 1986. – 52 p.
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  13. Степин Б.Д. Применение международной системы единиц физических величин в химии / Б.Д. Степи. – М.: Высшая школа, 1990. – 96 с.
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