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ВЛИЯНИЕ СЕРОВОДОРОДА НА СОКРАТИТЕЛЬНУЮ АКТИВНОСТЬ МИОКАРДА ПРАВОГО ЖЕЛУДОЧКА КРЫСЫ

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СОДЕРЖАНИЕ



стр.
СПИСОК СОКРАЩЕНИЙ3

ВВЕДЕНИЕ5

1    ОБЗОР ЛИТЕРАТУРЫ9

1.1 Роль H2S в межклеточной коммуникации и регуляции9

деятельности клетки


1.2 Взаимодействие NO, СО и H2S14

1.3 Физико-химические свойства H2S15

1.4 Образование и катаболизм H2S16

1.5 Токсичность и эндогенные концентрации H2S19

1.6 Физиологические и патофизиологические эффекты и механизмы20

действия H2S
1.7 Патологические процессы, связанные с метаболизмом H2S22

в организме


1.8 Роль H2S в сердечно-сосудистой системе23

1.9 Влияние H2S на сократимость миокарда26

1.10 Механизмы регуляции сократимости сердечной мышцы28

ЭКСПЕРИМЕНТАЛЬНАЯ ЧАСТЬ36

2     МАТЕРИАЛЫ И МЕТОДЫ36

2.1 Объект и методы исследования36

2.2 Растворы и фармакологические вещества38

3     РЕЗУЛЬТАТЫ И ОБСУЖЕНИЕ40

3.1 Эффекты донора Н2S-NaHS на сократимость изолированной полоски миокарда желудочка40

3.2 Роль К-каналов в отрицательном инотропном эффекте NaHS41

в изолированной полоске миокарда


3.3 Роль NO в эффектах H2S на сократимость миокарда43

3.4 Обсуждение результатов45

ВЫВОДЫ47

СПИСОК ИСПОЛЬЗОВАННЫХ ИСТОЧНИКОВ48
 
ВВЕДЕНИЕ
Последние достижения в физиологии в основном связаны с расшифровкой механизмов межклеточных взаимодействий и регуляции деятельности клетки. Кроме исследования классических посредников прошлый век ознаменовался открытием физиологической роли некоторых общеизвестных газообразных веществ. Оказалось, что сигнальную функцию в межклеточной коммуникации и во внутриклеточной регуляции выполняют известные своими токсичными эффектами газы: сероводород (H2S), монооксид углерода (СO) и оксид азота (NO) [Wang, 2004].
Существует ряд параметров, которым должен соответствовать потенциальный газообразный посредник: его молекулы должны существовать в виде газа и беспрепятственно проникать сквозь мембраны клеток, он должен иметь источник эндогенного синтеза и ферменты синтеза, должен участвовать в регуляции физиологических функций, иметь специфические клеточные и молекулярные мишени [Wang, 2002]. Все известные и исследованные на данное время газотрансмиттеры соответствуют этим параметрам.
В окружающей среде H2S образуется, главным образом, при разложении органических веществ. Также он обнаруживается в природном газе, нефти и выбросе вулканической серы [Gadalla et al., 2010]. Как и все газотрансмиттеры, H2S является небольшой молекулой, которая может проходить через клеточные мембраны без использования конкретных переносчиков. Большая часть сероводорода метаболизируется в тиосульфат и сульфат в ходе окислительного процесса в митохондриях. Только низкие уровни H2S могут быть превращены в цитозоле клетки в менее токсичные соединения [Chen et al., 2004; Stipanuk et al., 2010]. С целью поддержания сбалансированного уровня H2S, его метаболические продукты удаляются в течение 24 часов через почки, желудочно-кишечный тракт (ЖКТ) и легкие
 
[Furne et al., 2001]. В нормальных физиологических условиях, при отсутствии нарушений, связанных с метаболизмом H2S, его накопления не происходит. Это означает, что в нормальных для жизнедеятельности организма условиях, эндогенный H2S не токсичен для клеток.
Известно, что ткани млекопитающих могут образовать H2S через эндогенные системы синтеза, которые состоят в основном из двух ферментов
- цистатионин β-синтазы (ЦБС) и цистатионин γ-лиазы (ЦГЛ) [Kimura, 2010; Martin et al., 2010]. Аминокислота L-цистеин является основным субстратом для синтеза H2S. Последние исследования на людях показали, что H2S также может быть синтезирован в ЖКТ не только при помощи бактерий, но и из цистеина [Oh et al., 2006].
Физиологическая роль H2S показана для всех систем органов и тканей. Показаны эффекты H2S в нервной ткани [Abel, 1997; Kimura, 2000; Moore et al., 2003; Pushchina et al.,2011], в ЖКТ [Distrutti et al., 2006; Fiorucci, 2005; Hosoki, 1997; Schicho et al.,2006], в репродуктивной системе [Guzman et al.,2006; Liang et al.,2007; Srilatha et al.,2006], а также в сердечно-сосудистой системе [Bucci, 2009; Cheng et al., 2004; Dombkowski et al., 2004; Dombkowski et al., 2005; Fiorucci, 2006; Geng, 2004; Zhu et al., 2007].
Зависимость между уровнем содержания H2S в плазме крови, тканях и клетках и развитием различных заболеваний, протекторное действие при ишемическом повреждении жизненно важных органов позволяют считать этот газ важным звеном патогенеза многих заболеваний  центральной нервной и сердечно-сосудистой систем (ССС). Во многих исследованиях было показано, что физиологические эффекты H2S делают его идеально подходящим для защиты сердца, головного мозга, печени, почек и легких от повреждений при ишемии/реперфузии [Lavu et al., 2011].
Имеются данные о кардиопротекторной роли H2S, выражающейся в уменьшении повреждений миокарда в условиях ишемии/реперфузии в экспериментах in vitro и in vivo [Bian et al., 2006; Elsey et al., 2010; Geng et al.,
 
2004]. В единичных исследованиях показано, что H2S оказывает отрицательный инотропный эффект на сердце различных видов теплокровных животных и уменьшает длительность потенциала действия рабочих кардиомиоцитов [Geng et al., 2004; Sun et al., 2008].
Механизмы действия H2S малоизучены и включают, по разным данным, аденилатциклазную систему, различные ионные каналы в зависимости от вида животного [Sun et al., 2008; Xu et al., 2007; Yong, et al., 2008]. Установлено влияние H2S на тонус сосудов у всех классов позвоночных животных, включая как расслабление, так и усиление тонуса, что указывает на эволюционную древность H2S, как трансмиттера, и универсальность его действия [Dombkowski et al., 2004; Olson et al., 2006]. Данные о действии H2S на сократимость миокарда во многом фрагментарны и многие вопросы остаются открытыми.
Изучение механизмов влияния H2S на функциональную активность клеток может иметь значение не только с позиции фундаментального знания о принципах регуляции поведения клеток и механизмах внутриклеточной коммуникации, но и с точки зрения его терапевтической и профилактической значимости.
Основываясь на актуальности выявления роли газообразного посредника H2S в механизмах регуляции сократимости миокарда, нами была поставлена цель: исследовать эффекты сероводорода и выявить механизмы его действия на сократимость миокарда правого желудочка крысы (Rattus norvegicus).
Согласно поставленной цели нами были сформулированы следующие задачи:
      1. Исследовать дозозависимые эффекты донора сероводорода- гидросульфида натрия на сократимость миокарда правого желудочка крысы.
      2. Выявить роль К-каналов в инотропном эффекте донора сероводорода на миокард правого желудочка крысы.
 
      1. Исследовать роль системы оксида азота в эффектах сероводорода на сократимость желудочкового миокарда крысы.
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