Influence of various forms chitosan on redox processes in the liver and metabolic syndrome

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Influence of various forms chitosan on redox processes in the liver and metabolic syndrome
Kasimova Gulnorakhon Zulfikarovna, The assistent of Andijan State Medical Institute Sabirova Rikhi Ablukadirovna, The professor of Tashkent Medical Academy E-mail: evovision@bk. ru
Influence of various forms chitosan on redox processes in the liver and metabolic syndrome
Аbstract: When metabolic syndrome is develop the functional activity of the cytochrome P-450 in microsome of the liver decrease and develop disbalance in activity ferments of cycle tricarbone acid in mitochondria'-s. Keywords: metabolic syndrome, liver, microsomale and mithochondrial oxidation, hitozan.
The clinical significance of metabolic syndrome (MS), the combined framework syndrome is the presence of a whole range of risk factors, which are formed long before its development [1]. In clinical practice, MTS communications with impaired liver function is defined as the term & quot-diabetic steatosis& quot- (Canadian Diabetes Association. Canadian Diabetes Association 2003). Despite the importance of liver disorders in the development of metabolic syndrome, many aspects of the pathogenesis of this disease with hepatocytes, in particular molecular-cellular mechanisms remain unclear. Especially there is no clear clarity on the functional relationship systems, mitochondrial microsomal oxidation in the liver with the formation of MS.
Of particular interest is the study of the effect on the microsomal system, mitochondrial membrane protectors of the liver. There is some evidence that chitosan and its isoforms may positively affect the functional activity of the liver in acute and chronic hepatitis [2]. However, it remains unclear how does chitosan and its isoforms at the subcellular level in the liver with the formation MS.
The aim of this study was to investigate the microsomal, mitochondrial oxidation in the liver and the impact of various forms of chitosan on these processes.
Material and methods
Experiments were carried out on 60 male rabbits, body weight from 2050 to 3400 g. Metabolic syndrome in rabbits is caused by the method of S. A. Saidov [3]. To create a model of metabolic syndrome in animals trough added 5% sugar solution and mixed in a daily feed crystalline cholesterol of 250 mg/kg. body weight. Animals were subcutaneously injected insulin dose of 0.1 units/100 g., a day. The duration of 2 months. Animals were divided into 3 groups: Group 1 (intact) contained in natural vivarium conditions (12 rabbits). 2 group — called metabolic syndrome (12 rabbits). 3 group — correction of metabolic syndrome chitosan sulfate (12 rabbits) — 4 group — correction of metabolic syndrome, a form of nano chitosan sulfate (12 rabbits) — 5 group — comparison group, where the correction of the metabolic syndrome was performed Glyukofazh. Chitosan is a deacetylation product of chitin. The chemical structure of chitosan is a copolymer of D-glucosamine and N-acetyl-D-glu-cosamine. Chitosan is a universal sorbent capable of binding a huge range of substances of organic and inorganic nature, which defines the broadest possible application in human life. The study examined the effect of chitosan sulfate and Nana form synthesized based on chitosan Bombyx mori, represented by the Institute of Physics and Chemistry of the Academy of Sciences of Uzbekistan (Head of Laboratory, Dr. R. Y. Milusheva).
MS used for correction sulphate chitosan Chitosan obtained sulfation reaction medium in chlorosulfonic acid. Orally aqueous solution of chitosan and its sulfate form nano administered 25 mg/kg. over 20 days after receiving a model of metabolic
syndrome. Glucophage, according to the instructions of the drug was administered orally at the rate of — 7. 14 mg/kg. body weight.
Isolation of mitochondrial and microsomal fractions of the liver was carried out by differential centrifugation on a refrigerated centrifuge RL — 6 and VAC- 601. The contents of cytochrome P-450 in the microsomal suspension was determined with a spectrophotometer UV/VIS (LTD, China) by the method of Omura T., Sato R. [4]. The content of cytochrome b5 were determined after recovery experienced microsomal suspension samples by the addition of NADPN. Activity aminopyrine demethylase-N- method Bast A., Nordhoock J. [5], NADPH-cytochrome C-preductase method Wil-lians C. H., Kamin H. [7] and microsomal protein of O. N. Lowry [8].
Determination of liver mitochondrial enzyme activity of cytochrome oxidase (CCO) — polarographic method (in the LP-7 polarograph closed platinum electrode type Clarke), succinate dehydrogenase (SDH), succinate — cytochrome with reductase (SCR- ed.), Rotenone — insensitive NADPN — cytochrome with reductase and monoamine oxidase (MAO) spectrophotometric method [9].
Results and discussion
Studies have shown that after 2 months from the beginning of the simulation of the experimental MS in liver microsomes of cytochrome P-450 and B5 was significantly reduced by 28.6 (P & lt- 0. 01) and 17.2% (P & lt- 0. 05), respectively, compared with the intact group (Table 1). Activity-NADFH-cytochrome c reductase, aminopyrine demethylase-N-, anilingidroksilazy animals of this group decreased 2. 48- 1. 93 and 2. 14 times, respectively, compared to the intact group. This enzyme system plays an important role in metabolizing both endogenous (steroid hormones, cholesterol, fatty acids and bile acids, prostaglandins) or exogenous (xenobiotic majority) substrates, it is fully functional condition depends on the integrity of the endoplasmic reticulum membrane structures. Therefore, these results indicate a pronounced inhibition of microsomal oxidation in the liver and metabolic syndrome.
NADPH-cytochrome P-450 microsomes and mitochondria electron transport system is in constant renewal de novo protein enzyme complexes, metabolic activity which is largely dependent on the varying conditions of physiological and pathological processes in the cells [10]. Thus protein synthesis de novo enzyme complexes requires considerable use of ATP and NADPN. Mitochondrial and microsomal systems compete for NADPN connection with its use in the process of mitochondrial respiration in the free and conjugated with the operation of the cytochrome P-450 systems smooth endoplasmic reticulum [10- 11- 12- 13]. The development of many diseases is closely related with the damage of cell membranes, which leads to violation of their functional activity. Damage to the energy systems of the cell is unable to produce sufficient amounts of ATP and nukleotrifosfat [14]. In our studies, the study of mitochondrial
Section 5. Medical science
enzyme activity revealed a significant increase in the activity ofmono-amine oxidase (MAO) and cytochrome oxidase (CHO) for MS and 67. 55%, 58. 14% (P & lt- 0. 001) respectively compared to the intact group of animals (Table 2). At the same time, the activity of succinate dehydrogenase (DHN), succinate-cytochrome with reductase (SCC), rotenone-insensitive NADH-cytochrome with reductase was significantly reduced by 32. 92%- 42. 38%- 44. 19% (P & lt- 0. 001) respectively compared to intact group. Consequently, there are significant
shifts in the way ofusing oxidase O2 in the citric acid cycle. An imbalance in the system substrate oxidation in the citric acid cycle requires excessive consumption of NADPN. Inhibition of the activity of the liver MOS at the metabolic syndrome may be due to a deficiency of NADPH — as the main source for the functioning of NADPN — cytochrome c reductase in microsomes, as noted in our research. Currently offers a variety of methods to restore the microsomal oxidation processes, including chitosan derivatives (Table 1).
Table 1. — Effect of chitosan derivatives on microsomal oxidation in the liver of rabbits with the metabolic syndrome (M ± m)
Group Microsomes
P-450, nmol/mg. B5, nmol/mg. NADPH — cyt. -c. red., nmol/min/mg. AN, nmol НСНО/min/mg. AG, nmol ami-nofen/min/mg.
Intact 0. 985 ± 0. 030 0. 593 ± 0. 021 94.4 ± 8. 48 7.0 ± 0. 492 1. 09 ± 0. 06
MS 0. 703 ± 0. 024* 0. 491 ± 0. 004* 38.0 ± 2. 94* 3.6 ± 0. 28* 0. 51 ± 0. 026*
MS + chito-san sulfate 0. 775 ± 0. 033* 0. 484 ± 0. 024* 49.6 ± 3. 27* 4.4 ± 0. 30* ** 0. 57 ± 0. 024*
MS + chitosan sulfate nano 0. 837 ± 0. 026* ** *** 0. 518 ± 0. 019* 53.8 ± 2. 48* ** *** 5.5 ± 0. 177* ** *** 0. 67 ± 0. 034* ** ***
MS + glyuko fazh 0. 706 ± 0. 032* 0. 497 ± 0. 021* 45.7 ± 1. 76* 3.6 ± 0. 28* 0. 54 ± 0. 032*
Note: * - differences with respect to the data of the control group significant (* - P & lt- 0. 05- ** - P & lt- 0. 01- *** - P & lt- 0. 001).
Investigating the effects of the three forms of chitosan — chitosan sulfate and nano forms on monooxygenase system showed a significant increase in the content of cytochrome P-450. When administered chitosan and its sulfate form nano levels of cytochrome P-450 exceeded by 10.2 and 19.1% (P & lt- 0. 05 and P & lt- 0. 05, respectively) compared to those animals with MS parameters. Chitosan Sulfate and its nano form of significant changes in the content of
cytochrome B5 causes. Chitosan Sulfate significantly to 30. 45% (P & lt- 0. 01) increases the activity of only NADPN-cytochrome C-ed. compared to the untreated group. At the same time form nano chitosan sulfate increases more significantly as activity NADPN-cytochrome c-ed and aminopyrine demethylase-N-, anilingidroksilazy 41.6%, 53.6%, 31.7% (P & lt- 0. 001), respectively, compared with the untreated group (Table 2).
Table 2. — Effect of chitosan derivatives on mitochondrial oxidation in the liver of rabbits with the metabolic syndrome (M±m)
№ Group МАО, DHN, CCO-red., РН-cyt. С-red., CHO,
nmol/min/mg. nmol/min/mg. nmol/min/mg. nmol/min/mg. nmol/min/mg.
1 Intact 23.0 ± 0. 09 127.8 ± 4. 144 120.2 ± 3. 35 55.1 ± 2.0 120.3 ± 2. 63
2 MS 38.6 ± 1. 34* 85.7 ± 1. 53* 69.3 ± 2. 3* 30.7 ± 1. 39* 190.2 ± 8. 49*
3 MS + chitosan sulfate 35.1 ± 1. 41* 97.2 ± 3. 70* 89.6 ± 3. 48* ** 40.2 ± 1. 67* 150.4 ± 7. 54* **
4 MS + chitosan sulfate nano 28.5 ± 1. 47* ** *** 116.2 ± 2. 54* ** *** 103.8 ± 4. 60* ** *** 46.8 ± 1. 84* ** *** 133.6 ± 6. 25* ** ***
5 MS + glyuko fazh 37.2 ± 1. 32* 90.7 ± 3. 37* 70.5 ± 4. 23* 33.8 ± 1. 44* 188.3 ± 9. 83*
Note: * - differences with respect to the data of the control group significant (* - P & lt- 0. 05- ** - P & lt- 0. 01- *** - P & lt- 0. 001).
A study comparing the action of the drug — Glucophage has shown that he is not sufficiently active to enhance the functional activity of microsomes and mitochondria. Also, no significant differences in the study Glyukofazh action on the activity of mitochondrial enzymes. Glucophage is widely used as a means of correction dismet-abolic disorders in MS development. Perhaps it is not related to the effect of the influence on the processes oxidase and oxygenase way of oxidation. Unlike Glyukofazh chitosan derivatives and chitosan nano largely affect the activity of mitochondrial enzymes oxidation. This nano chitosan has a greater influence on the process of reducing the activity of MAO and CCO and increase the reaction rate of LDH, SO tsit. s. red. RN-tsit. s. red than chitosan sulfate, respectively — 18.8%, 11.2% (P & lt- 0. 05) and 19. 5- 15. 8- 16.4% (P & lt- 0. 05).
It is believed that with the improvement of the functional activity of the enzymes oxidase oxidation way, helped to improve the functioning of the citric acid cycle enzymes, transport of electrons
and protons along the respiratory chain to its final link — CCO. In this case the electron acceptor is O2 and the protons, which is formed in the reduction of H2O. This oxidation pathway is known to conjugate and ADP phosphorylation and the resynthesis ofATP in the mitochondria NADPN. It should be noted that excessive CCO activity may be an important factor in the formation of the hydroxyl radical, which is a strong oxidizing agent active center of the cytochrome P-450 enzyme activity inhibition of microsomal oxidation.
Thus, studies have shown that the development of MS pronounced inhibition observed functional activity of cytochrome P-450 system and liver microsomes imbalance in activity of the enzymes of the tricarboxylic acid cycle in mitochondria.
Chitosan derivatives, to a greater extent chitosan nano, positive impact on the recovery of impaired activity of enzyme systems microsomes and mitochondria, it is possible to believe, it is one of the reasons for the decline of MS factors in experimental animals.
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Kosimov Azam Azimovich, Ph. D in Medicine, Scientific Research Institute Traumatology and Orthopedics of the Republic of Uzbekistan, Tashkent city E-mail: azamrefracture@mail. ru
Roentgenologic description of repeated fractures of forearm bones in children
Аbstract: The successful development of traumatology orthopedics in the last decade has not solved some of the problems which, in particular, to repeat the treatment of bone fractures in children. Nonunion and refracture are a common complication of bone fractures and trauma require attention. The main method of diagnosis is X-ray examination, which not only allows us to determine the nature of the displacement of bone fragments, but also plays a major role in the evaluation of treatment results. Keywords: children, refracture, X-ray examination, forearm.
According to the literature it is known that bone fractures in children ranged from 52% to 58% [2- 4- 5]. In 5.2% of them concern to the refractures in children [1- 7- 8]. Repeated bone fractures in children are some of the serious injuries and at the same time, diagnosis and treatment are accompanied by some difficulties. The main diagnostic method of re-fracture is X-ray examination, which not only allows us to determine the nature of the displacement of bone fragments, but also plays a major role in the evaluation of the results of surgical treatment. Diagnosis of this pathology is challenging and misdiagnosis can lead to different consequences and complications.
Incomplete gathering of the anamnesis in children without finding-out of the reasons of occurrence of a trauma, an establishment of terms of occurrence concerning primary fractures ofbones with finding-out of a way of treatment after primary fractures can complicate a choice of an adequate way of treatment in case of re-fracturein the same place.
The only prevention of complications is a detailed description of these radiographs. According to available literature, complications ofvarious character such as re-refracture occur in cases of 1. 4−1.7% and pseudoarthrosis — in 2. 4−2.6% cases [3- 6- 8]. In the course of studying this problem, we realized that nowhere and in any literary source is not given x-ray data and defined refracture problems in children. The relation of the radiological signs of re-fracture by
osteoreparative processes is not defined and not demonstrated the status of callus during the re-fracture.
Based on the abovementioned, we have put the purpose as to explore and explain the radiological signs of refractures, depending on the stage of the regeneration of the primary fracture.
Materials and methods
In Children Traumatology Department of the Scientific Research Institute of Traumatology and Orthopedics of MH of the Republic of Uzbekistan, were examined and treated 109 children with refracture (without concomitant somatic diseases), during the period from 2000 to 2014. From them 77 (70.6%) were boys and 32 (29.4%) girls. The distribution of children according to the localization was as follows: with diaphyseal refracture were 91 (83.5%), in the proximal part — 7 (6.4%) cases and in the distal portion were 11 (10.1%) children. Also, children are separated depending on the fracture frequency in the same segment, i. e. twice (refracture) in 91 (83.5%), children, re-refracture or many times (more than three times) — in 18 (16.5%) patients. Depending on the type of injury was observed the home injury — in 57 (52.3%) children, street trauma — in 44 (40.3%) and during sport activities — in 8 (7.4%) children. 66 (60.5%) patients treated conservatively and another 43 (39.5%) patients by surgery way, from them. 43 (39.5%) children in 24 (22.0%) cases was used the combined method of osteosynthesis using the Ilizarov'-s apparatus, in 16 (14.7%) — intramedullary

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