Резюме
Звягінцева Т.Д., Гаманенко Я.К., Савохіна М.В., Деримедвідь Л.В., Кніженко І.Б. Доцільність застосування препарату мелатоніну у пацієнтів з хронічними ерозіями шлунку з позицій порушення функціональної морфології ентерохромаффінних клітин дифузної ендокринної системи.
В даний час доведено, що порушення в міжклітинних механізмах ентерохромаффінних (ЕС) - клітин пов'язане з порушенням функціональної морфології дифузної ендокринної системи (ДЕС). Залежно від рівня концентрації та функціонального стану ЕС-клітин, мелатонін проявляє багатофазову дію в механізмі виникнення хронічних ерозій шлунку (ХЕШ). Дані факти дозволили припустити, що ступінь тяжкості клінічного перебігу ХЕШ може корелювати з вираженістю порушень продукції мелатоніну, що і послужило підставою для даного дослідження. Результат морфологічного дослідження показав, що зниження цитопротективних властивостей і регенераторної функції слизової оболонки шлунку (СОШ) обумовлено гіперплазією ЕС-клітин, що створює умови для розвитку запалення і прогресування ХЕШ. Зміна функціональної морфології ДЕС, свідчить про патогенетичну роль порушень місцевого гормонального гомеостазу у формуванні запально-дистрофічних процесів в СОШ при ХЕШ. Встановлено, що розвиток ХЕШ супроводжується зміною морфофункціональних показників ЕС-клітин. При використанні в комплексній терапії мелатоніну показники функціональної морфології ЕС-клітин відновлюються в значно більшій мірі, ніж при застосуванні стандартної антихелікобактерної терапії, що свідчать про доцільність використання препаратів мелатоніну в комплексній терапії ХЕШ.
Ключові слова: хронічні ерозії шлунку, дифузна ендокринна система, EС-клітини, мелатонін.
Резюме
Звягинцева Т.Д., Гаманенко Я.К., Савохина М.В., Деримедведь Л.В., Книженко И.Б. Целесообразность применения препарата мелатонина у пациентов с хроническими эрозиями желудка с позиций нарушения функциональной морфологии энтерохромаффинных клеток диффузной эндокринной системы.
В настоящее время доказано, что нарушение в межклеточных механизмах энтерохромаффинных (ЕС) – клеток связано с нарушением функциональной морфологии диффузной эндокринной системы (ДЭС). В зависимости от уровня концентрации и функционального состояния ЕС -клеток, мелатонин проявляет многофазовое действие в механизме возникновения хронических эрозий желудка (ХЭЖ).  Данные факты позволили предположить, что степень тяжести клинического течения ХЭЖ может коррелировать с выраженностью нарушений продукции мелатонина, что и послужило основанием для данного исследования. Результат морфологического исследования показал, что снижение цитопротективных свойств и регенераторной функции слизистой оболочки желудка (СОЖ) обусловлено гиперплазией ЕС-клеток, что создает условия для развития воспаления и прогрессирования ХЭЖ. Изменение функциональной морфологии ДЭС, свидетельствует о патогенетической роли нарушений местного гормонального гомеостаза в формировании воспалительно-дистрофических процессов в СОЖ при ХЭЖ. Установлено, что развитие ХЭЖ сопровождается изменением морфофункциональных показателей ЕС-клеток. При использовании в комплексной терапии мелатонина показатели функциональной морфологии ЕС-клеток восстанавливаются в значительно большей мере, чем при применении стандартной антихеликобактерной терапии, что свидетельствует о целесообразности использования препаратов мелатонина в комплексной терапии ХЭЖ.
Ключевые слова: хронические эрозии желудка, диффузная эндокринная система, ЕС-клетки, мелатонин.
Summary
Zviagintseva Т.D., Gamanenko Ya.K., Savokhina М.V., Derymedvid L.V., Knizhenko I.B. Expediency of melatonin usage in patients with chronic gastric erosions from the position of the functional morphology disorders in enterochromaffin cells of the diffuse endocrine system.
At present, it has been proved that the disorder in the intercellular mechanisms of enterochromaffin (EC) cells is caused by the functional morphology of the diffuse endocrine system’s (DES) disorder. Depending on the concentration level and EC-cells functional status, melatonin exhibits multi-phase action in the mechanism of chronic gastric erosions (CGE). Based on these facts we can suggest that the severity of the CGE clinical course is correlated with the severity of melatonin production disorder. The results of morphometric study have showed that the decrease in cytoprotective properties and regenerative functions of the gastric mucosa (GM) is caused by EC-cell hyperplasia, which creates conditions for the inflammation development and CGE progression. Changes in DES functional morphology indicate the pathogenetic role of hormonal homeostasis local disorders in the formation of inflammatory and degenerative processes in CGE. It has been established that CGE development is accompanied by changes of EC-cells morphofunctional indices. When used in complex melatonin therapy, indicators of EC-cells functional morphology are restored to a much greater extent than with the standard therapy of H. pylori, which suggests the expedient use of melatonin medication in the complex treatment of CGE.
Key words: chronic gastric erosions, diffuse endocrine system, ЕС-cells, melatonin.

УДК 616.33-002-092-07-085.243+615.276

UDC 616.33-002-092-07-085.243+615.276

EXPEDIENCY OF MELATONIN USAGE IN PATIENTS WITH CHRONIC GASTRIC EROSIONS FROM THE POSITION OF THE FUNCTIONAL MORPHOLOGY DISORDERS IN ENTEROCHROMAFFIN CELLS OF THE DIFFUSE ENDOCRINE SYSTEM

Т. D. Zviagintseva, Ya. K. Gamanenko, М. V. Savokhina,

L. V. Derimedved’, I. B. Knizhenko

Национальный фармацевтический университет (Харьков) 61002, г. Харьков, ул. Пушкинская, 53
National University of Pharmacy (Kharkov) Ukraine, 61002, Kharkov, 53, str. Pushkinska

Харьковская медицинская академия последипломного образования 61176, г. Харьков, ул.Корчагинцев, 58

Kharkov medical academy of Postgraduate Education 61176, Kharkov, 58 str.Korchahyntsev

derimedved@mail.ru

marinadoc10@gmail.com

In recent studies we have noticed a clear tendency to the increase in frequency of erosive lesions of gastric mucosa (GM) and its complications [9]. In domestic and foreign publications the ulcers and erosions formation mechanisms are mainly discussed but not chronic processes, and therefore the questions on pathogenesis of chronic gastric erosions (CEG) have not been fully elucidated, and the causes of the pathological process has not been fully studied [4].

At present it has been proved that disorders in the intercellular mechanisms of enterochromaffin (EC cells) is due to malfunctional morphology of the diffuse endocrine system (DES). The processes of EC-cells renewal are regulated by DES, therefore its functional morphology disorders could lead to inflammatory changes in GM and CGE [1, 7, 11].

The nature of DES’s influence on cell homeostasis in EC-cells in diseases of the gastrointestinal tract (GIT) is being studied. The solution to this problem is important because basic processes of damage are associated with malfunctioning of the regenerative GM caused by hyperplasia of EC cells that creates the conditions for the development and progression of CGE [8, 12].

The purpose of the research is to study the causes of CGE based on morphological and functional indicators of EC-cells activity GM and DES functioning.

Materials and methods. We have studied the morphological and functional features of the EC-cells in 64 patients with acute CGE, aged 21 to 40 years (average age was 28.3 +1,75 years), including 49 men (76,6%) and 15 women (23,4%).

The patients were divided into 2 groups: the first group consisted of 15 (44,1%) patients with CGE diagnosed for the first time, the second group included 34 (55,9%) patients with CGE to 5 years (of which - 15 people made a comparison group). The patients in both groups were adequately matched by age, endoscopic, morphological and histochemical characteristics.

The patients in I and II group underwent of H. pylori therapy on the traditional pattern with the use of lansoprazole, 30 mg twice a day, amoxicillin 1000 mg twice a day, clarithromycin 500 mg twice a day and melatonin medication - melaksene 6 mg per night for 12 days. The patients from the comparison group received traditional 1st line eradication scheme.

Addition of melatonin medication to the standard CGE treatment patterns is due to the fact that a number of researchers [1, 5, 6] have noted the presence of gross disorders of melatonin production in patients with peptic ulcer (PU) of the duodenum, which reduced in remission, but did not completely disappear. Based on the desynchronosis fact of melatonin production in patients with remission, many scientists believe that a change in melatonin production may be a viable element of acute duodenal ulcer [1, 3, 6]. Melatonin helps optimize gastrointestinal mucosa cell proliferation by enhancing production of prostaglandins E2 (Pg2E), participating in gastroprotection [3, 10, 12]. Antioxidant properties of melatonin realized by binding of free radicals (singlet oxygen, superoxide anion radical), hydroxyl radicals and antioxidant system stimulation (first of all - glutathione peroxidase) are of interest, too. Unlike other antioxidants, it is able to penetrate into any cell in any part of the body and exert a specific protective effect on the nucleus [1, 3].

Both groups of CGE patients have been examined in the dynamics on a common program that included clinical research methods, fibrogastroduodenoscopy (FGDS), general morphological and histochemical examination determining morphological and functional activity of the EC-cells. To assess the changes in the values of the EC cells morphofunctional activity, we compared them with a control group of practically healthy people. Morphological images coming through the optical system of Nikon Eclipse microscope (magnification 400, lens 40, eyepiece 10, filter 0,8) have been recorded with a digital color camera Nikon DXM1200 mounted in the tube of microscope and transferred to Pentium-4 computer with subsequent microphotography.

Material for histological examination was taken from the antrum of the stomach. 3-4 biopsies of GM obtained by pinch from the edges of chronic erosion were fixed in 10% neutral formalin, dehydrated in alcohol, embedded in paraffin, and then serial sections 4-5x10ˉ6 m thick were prepared. To determine Helicobacter pylori (HP) we used histological research method with coloring on the Shubich method and toluidine blue. The degree of GM HP insemination was assessed by classification criteria proposed by L.I Aruin [2].

For EC cells nuclei staining hematoxylin-eosin and toluidine blue were used. In metachromatic staining light absorption maximum of metachromatically colored formation differs from the absorption maximum of the dye. When stained with toluidine blue, EC cells grains are colored purple-red color, while other formations perceive the original blue dye. Survey preparations stained with hematoxylin-eosin were used to assess the overall state of the EU-cells.

For a more in-depth study of nuclear structure of the EC-cells an enterochromaffine reaction was used, characterized by the identification of grains in the cytoplasm. When applying this method, enterochromaffin cells are colored dark red. As concerns the chemical composition of the grains, according to the latest research, the most probable is presence of the 5-hydroxytryptamine in them [8].

To determine the level of morphofunctional activity of EC-cells the following formulae were used:                                                          

Indicators characterizing cellular homeostasis of EC-cells GM of antrum had been determined before the complex therapy treatment and after 12 ± 4 days of treatment.

The results of the study were mathematically calculated using a statistical software package Excel on a PC IBM Pentium-4.

Results and discussion. Depending on the concentration level and the functional state of the EC-cells melatonin exhibits multi-phase action in the mechanism of CGE origin. These facts suggest that the severity of the CGE clinical course may vary depending on the morphological and functional disorders of EC-cells of GM.

Morphologic study of gastrobiopsy in patients of I and II groups showed that fibrosis areas disappeared, the gastric glands clearance, demarcation and infiltration of perierosive zone decreased. Therefore, we can state the morphological confirmation of CGE epithelialization. However, in the comparison group morphological picture in 53.4% of patients remained virtually unchanged, and 27% had increased polymorphonuclear cell infiltration in GM.

Controlling the eradication, we noted that Hp eradication was achieved in 82,3%, in patients from group I, in group II it was higher than 85%, while in the control group - 74.3%. 4 weeks after the therapy was finished, the complete eradication was observed in all patients, except for the comparison group, in which we noticed insemination of I degree once again. Thus, acombination therapy with duration of 12 days course, has a significant effect of anti-H. pylori, persisting for 1 month after the end of treatment.

During the comprehensive study of morphometric parameters in patients of group I we have found out that EC-cells are uniformly distributed throughout the whole layer of GM among the glands’ epithelial cells. The patients in group II showed increased destructive processes with increasing strength of inflammation, until death (apoptosis) of endocrine cells and adjacent epithelial cells. At the same time, intensive excretion of secretory material from the cell by dissolving the secretory granules and the secret diffusion through the cell membrane has been noticed. Clear predominance of endocrinocytes with polygonal nucleus and highly vacuolated cytoplasm has been established

Consequently, we observed a high level of functional activity of the remaining small number of endocrine cells showing signs of depletion and apoptosis. Judging by GM histology in patients of group II, it can be assumed that the sharp increase in demand for the synthesis of melatonin leads to the exhaustion of the morphofunctional ability of EC-cells and the development of accelerated distress apoptosis. This may be caused by the lack of "protection" in the form of endogenous melatonin, which is the basis for the use of exogenous melatonin in complex CGE treatment, as optimal pathogenetic therapy should be aimed not only at the eradication of bacteria, but should also influence the pathogenetic chronicity mechanisms of the process itself.

To assess the effectiveness of the complex therapy influence on the morphological and functional condition of DES cells in patients with CGE we have analyzed the dynamics of EC-cells indicators only in patients of group II and in the comparison group, as morphofunctional pattern in group I was identical to the control group indicating a sufficient melatonin synthesizing level of EC-cells.

In patients of group II EC cells are more compact. We have noticed EC-cells with large nucleus and eosinophilic cytoplasm which also testifies to active synthesis of neurotransmitters. Apoptosis is much less expressed than in the comparison group. This can be explained by the fact that in combination therapy the required level of melatonin synthesizing function in EC-cells is formed ”satisfying the needs” of the body by the input of melatonin (Fig. 3).

After the course of treatment the patients in group II have shown the recovery of morphofunctional activity of  EC-cells (Table 1).

Table 1.

Key morphometric parameters of enterochromaffin EC cells in patients with CGE up to 5 years, and in the comparison group

Note: * p <0,05, ** p <0,01, *** p <0,001 - compared with similar parameters before and after treatment in patients of group II.

ˇp> 0,05 - when comparing similar parameters before and after treatment in the comparison group.

Conclusions:

1. The results of morphological research have shown that the decrease in cytoprotective properties and regenerative function of GM is caused by hyperplasia of EC cells that creates conditions for the development of inflammation and progression of CGE. Changes in functional morphology of DES indicate the pathogenetic role of local hormonal homeostasis disorders in the formation of inflammatory and degenerative processes in GM in CGE.

2. The conducted research has proved that the development of CGE is accompanied by changes in morphofunctional indices in EC cells. On the one hand, this is the evidence of the complex endogenous mechanisms of CGE development, on the other hand, it opens new possibilities to correct pathological processes development and to optimize their treatment.

3. When using melatonin in complex therapy, the indicators of EC cells functional morphology are restored to a much greater extent, than in standard therapy of anti-H. pylori.

4. The obtained results indicate the expediency of using melatonin medications in complex CGE therapy.

Национальный фармацевтический университет (Харьков) 61002, г. Харьков, ул. Пушкинская, 53
National University of Pharmacy (Kharkov) Ukraine, 61002, Kharkov, 53, str. Pushkinska
Харьковская медицинская академия последипломного образования 61176, г. Харьков, ул.Корчагинцев, 58 Kharkov medical academy of Postgraduate Education 61176, Kharkov, 58 str.Korchahyntsev

derimedved@mail.ru

marinadoc10@gmail.com

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