Автореферат и диссертация по медицине (14.03.03) на тему:Реактивность макрофагальной системы при фибропластических процессах воспалительного генеза: оценка, механизмы регуляции и патогенетическое значение

ВВЕДЕНИЕ

раздел 1. обзор литературы.

часть 1.1. общая характеристика фибропролиферативных процессов.

глава 1.1.1. современные представления о патогенезе гепатофиброза.

1.1.1.1.1. Продукция соединительной ткани при развитии гепатофиброза.

1.1.1.1.1.1. Общая характеристика СТ при гепатофиброзе.

1.1.1.1.1.2. Механизмы продукции соединительной ткани.

1.1.1.1.1.3. Клетки-продуценты соединительной ткани.'.

1.1.1.1.2. Механизмы фиброгенного действия макрофагов.

1.1.1.1.2.1. Воспалительная Мн-Мф инфильтрация как источник фиброгенных факторов.

1.1.1.1.2.2. Трансформирующий фактор роста-бета (ТФР-Р).

1.1.1.1.2.3. Фактор роста соединительной ткани (СТОЙ).

1.1.1.1.2.4. Фактор роста, происходящий из тромбоцитов (РОСЕ7).

1.1.1.1.2.5. Факторы роста фибробластов (РвР).

1.1.1.1.3. механизмы антифиброзного действия Мф.

1.1.1.1.3.1. Металлопротеиназы и тканевые ингибиторы металлопротеиназ.

1.1.1.1.3.2. Фактор некроза опухоли-а (ТНФ-а).

1.1.1.1.3.3. Интерлейкин-1 бета (ИЛ-1 р).

1.1.1.1.3.4. Интерферон-гамма (ИФН-у).

1.1.1.1.4. молекуляр11ые механизмы функционального антагонизма провоспалительных цитокинов и ТФР-р.

глава 1.1.2. современные представления о фиброзном процессе прш атеросклерозе.

1.1.2.2.1. Основные варианты атеросклерот ического повреждения сосудов.

1.1.2.2.2. атеросю1еротическое поражение сосудов как хронический воспалительный и фибропластический процесс.

1.1.2.2.3. про- и антивоспалительные/фиброгенные цитокины в атерогенезе.

глава 1.1.3. современные представления о роли мф в развитии/ нефросклероз а.56.

1.1.3.3.1. Значение Мн-Мф инфильтрации.

1.1.3.3.2. Продуценты СТ при развитии нефросклероза.

1.1.3.3.3. Деградация СТ при развитии нефросклероза.

часть 1.2. современные представления о реактивности мф и смф в. целом.

глава 1.2.1. классическая активация и гиперреактивность мф.

1.2.1.1. Общая характеристика примирования и активации Мф.

1.2.1.1.1. История и общаяхарактеристика.

1.2.1.1.2. Критерии активации.

1.2.1.2. Молекулярные механизмы активации.

1.2.1.2.1. Примирование.

1.2.1.2.2. Разрешающий сигнал.

1.2.1.3. Гиперреактивность Мф на уровне популяций.

глава 1.2.2. гипореактивность мф.!.

1.2.2.1. эндотоксиновая толерантность.

1.2.2.1.1. История и общая характеристика.

1.2.2.1.2. ЛПС-толерантный фенотип Мф.

1.2.2.1.3. Молекулярные механизмы.

1.2.2.2. Альтернат ивно активированные Мф.

1.2.2.2.1. Игё-цитокин-индуцированные ааМФ.

1.2.2.2.2. ааМф II типа.

1.2.2.2.3. М1-М2 поляризация.

часть 1.3. липидный обмен при воспалении и его участие в регуляции реактивности мф.

глава 1.3.1. изменения липидного обмена при воспалении.

1.3.1.1. Триглицериды.

1.3.1.2. Mebajioi iai ный путь.

1.3.1.3. холеслерин.

глава 1.3.2. оксистеролы, lxr и реактивность мф.

1.3.2.1. роль оксистеролов и lxr в липидном обмене при воспалении.

1.3.2.2. оксистеролы в LXR-независимой регуляции.

1.3.2.3. оксистеролы и lxr в регуляции функций мф при воспалении.

раздел 2. собственные результаты.

часть 2.1. материалы и методы исследования.

часть 2.2. реактивность мф при хроническом воспалении и поствоспалительном фиброзе.108«

глава 2.2.1. реактивность мф при развитии гепатофиброза.

2.2.1.1. СМФ ПРИ РАЗВИТИИ ССЬ4-ГЕПАТОФИБРОЗА.

2.2.1.1.1. Общая характеристика функционального состояния Мф при СС14-гепатофиброзе.

2.2.1.1.1.1. Макрофаги печени.

2.2.1.1.1.2. Внепеченочные отделы СМФ.

2.2.1.1.1.2. Реактивность Мф при СС14-гепатофиброзе.

2.2.1.1.2.1. Макрофаги печени.

2.2.1.1.2.2. Внепеченочные отделы СМФ.

2.2.1.1.2.3. СМФ в целом.

2.2.1.2. СИС1ЕМНАЯ ЭНДОТОКСИНЕМИЯ КАК ФАКТОР ГИПОРЕАКТИВНОСТИ СМф|пРИ CCL4

ГЕПА1ОФИБРОЗЕ.

2.2.1.3. функциональное COCI ояние и реактивность МФ при затяжном вирусном гепатите человека.Г.

2.2.1.3.1. Показатели функциональной активности СМФ при лечении вирусного гепатита продигиозаном.

2.2.1.3.2. Показатели терапевтической эффективности продигиозана при лечении больных вирусным гепатитом.

2.2.1.4. ЛИПИДНЫЙ ПРОФИЛЬ СЫВОРОТКИ КРОВИ ПРИ ХРОНИЧЕСКОЙ СИСТЕМНОЙ ЭНДО'ГОКСИНЕМИИ.

глава 2.2.2. гиперлипидемия и эндотоксинемия как факторы реактивности смф при атерогенезе.

2.2.2.1. эндотоксин-липопрот еиновые комплексы в атерогенезе.

2.2.2.1-.1. Формирование эндотоксин-липопротеиновых комплексов при гиперлипидемиях.

2.2.2.1.2. Активность лецитин-холестерин-ацилтрансферазы (ЛХАТ) при формировании эндотоксин-липопротеиновых комплексов.

2.2.2.1.3. Клиренс эндотоксин-ЛПНП комплексов.

2.2.2.1.4. Аккумуляция комплексов эндотоксин-липопротеины низкой плотности в макрофагах.

2.2.2.1.5. Связывание и включение 1251-ЛПНП и 1251-ЛПНП-ЛПС в эксплантаты аорты крыс.

2.2.2.1.6. Влияние хронической эндотоксинемии на содержание ХС и ЭХС в артериальной ткани

• у мышей С57В1/6, находившихся на ХС диете.

глава 2.2.3. характер гранулематозного воспаления, вызванного фиброгенным и нефиброгенным материалом при генетически обусловленных различиях реагирования омгр и реактивности^ смф.

2.2.3.1. Реакция СМФ на зимоза! i у mi шредных мышей разных линий.

2.2.3.1.1. Зимозан-индуцированные гранулемы.

2.2.3.1.2. Содержание коллагена.

2.2.3.1.3. Численность клеток Купфера (КК), захват ими коллоидного угля.

2.2.3.1.4. Содержание ПГЕ2 в ткани печени.

2.2.3.2. Реакция СМФ на двуокись кремния у инбредных мышей разных линий.

2.2.3.2.1. 8Ю2-индуцированные гранулемы.

2.2.3.2.2. Содержание коллагена.

2.2.3.2.3. Численность клеток Купфера (КК), захват ими коллоидного угля.

2.2.3.2.4. Содержание ПГЕ2 в ткани печени.'.

2.2.3.3.5. Миелопоэз в костном мозге.

глава 2.2.4. липид-зависимые механизмы регуляции реактивности смф в фиброгенезе.i.

2.2.4.1. Исследование липид-зависимых механизмов регуляции реактивности СМФ в фиброзном процессе in vivo.

2.2.4.1.1. Аккумуляция триглицеридов, ХС, эфиров ХС и оксистеролов в ткани печени при СС14- или зимозан-индуцированном воспалении.

2.2.4.1.2. Влияние ХС-диеты, ингибиторов ОМГ-редуктазы и мевапоната на содержание коллагена в ткани печени при Z- и СС14-индуцированным гепатите.

2.2.4.1.3. Влияние ХС-диеты на экспрессию мРНК ТФР -ß в ткани печени при Z- и CCI4-индуцированным гепатите.

2.2.4.1.4. Влияние ХС-диеты на реактивность СМФ.

2.2.4.1.4.1. Действие ХС диеты на ЛПС-индуцированный уровень ФНО-а в сыворотке крови.

2.2.4.1.4.2. Действие ХС диеты на Z-индуцированный гранулемогенный ответ и численность клеток Купфера.

2.2.4.1.4.3. Действие ХС диеты на Z-индуцированный миелопоэтический ответ в костном мозге.

2.2.4.2. Исследование липид-зависимых механизмов регуляции реактивности СМФ в фиброзном процессе in vitro.

2.2.4.2.1. Действие ХС и ОС на экспрессию и продукцию провоспалительных цитокинов в Мф.

2.2.4.2.2. Действие ХС, ОС и других агонистов ядерных гормональных рецепторов на продукцию ТФР-ß вМф.

2.2.4.2.2.1. Действие ХС, ОС и других агонистов ядерных рецепторов на продукцию ТФР-ß в рекрутированных Мф, культивируемых в присутствии или отсутствии мевалоната и ЛИС.

2.2.4.2.2.2. Действие ЛИС и мевалоната на продукцию ТФР-ß в культуре рекрутированных

Мф, полученных у мышей, находившихся на ХС диете.

2.2.4.2.2.3. Продукция ТФР-ß в культуре толерантных к эндотоксину перитонеальных Мф и влияние на нее мевалоната.

раздел 3. обсуждение результатов.

часть 3.1. реактивность мф и уровень фиброгенного ответа при гранулематозном воспалительном процессе у мышей разных линий".

глава 3.1.1: реактивность мф системы при si02- и z-индуцированном воспалении.

3.1.1.1. Динамика Si02- и Z-индуцированного гранулемогенеза и популяции КК.

3.1.1.1.1. Основные отличия Si02- и Z-гранулемогенеза.:

3.1.1.1.2. Биодеградабельность частиц Z и Si02 и длительность стимуляции Мф.

3.1.1.1.3. Инфильтративная реакция и изменение численности Мф печени в ответ HaZ и Si02.

3.1.1.2. инфильтрлтивный, провоспллительный и фиброгенный ответ НА S1O2 и Z: механизмы различий.

3.1.1.2.1. Дискордантность инфильтративного и фиброгенного ответов при силикозе.

3.1.1.2.2. ScR и апоптоз при Si02- и Z-гранулематозе - роль в фиброгенезе.

3.1.1.2.3. Провоспапительная активность Мф под действием Z и Si02.

3.1.1.2.4. Миелопоэз при Z- и 8Ю2-гранулемогенезе.

3.1.1.2.5. Генерация ПГЕ2 при Z- и 8Ю2-гранулематозе.

3.1.1.2.6. Фенотип Мф, ассоциированный с 8Ю2-фиброзом (8Ю2-гранулематоз).

3.1.1.2.7. Фенотип Мф ассоциированный с отсутствием фиброза (Z-гранулематоз).

глава 3.1.2. межлинейные различия реактивности мф и выраженность фиброза при 8102-индуцир0ванн0м гранулемогенезе.

3.1.2.1. Межлинейные различия Si02- и Z-индуцированного гранулемогенеза.

3.1.2.1.1. Универсальный характер межлинейных различий гранулемогенеза.

3.1.2.1.2. Механизмы межлинейных различий гранулемогенеза.

3.1.2.1.1. Роль Th 1 -М1 /ТИ2-М2 поляризации.

3.1.2.1.2. Роль активности миелопоэза.

3.1.2.1.3. Роль миелопоэз-независимых механизмов.

3.1.2.1.4. Роль мевапонатного биохимического пути.

3.1.2.2. Межлинейные различия 8Ю2-индуцированного фиброгенеза.

3.1.2.2.1. Механизмы межлинейных различий.

3.1.2.2.1.1. Рол ь Th 1 -М1 /ТЬ2-М2 поляризации.

3.1.2.2.1.2. Роль миелопоэза.

3.1.2.2.1.3. Роль апоптоза.

3.1.2.2.1.4. Роль мевапонатного биохимического пути.

часть 3.2. реактивность мф при ссь4-индуцированном гепатофиброзе и затяжном вирусном гепатите человека.

глава 3.2.1. механизмы функциональной перестройки печеночного и внепеченочных отделов смф при ссь4-гепатофиброзе.216 <

3.2.1.1. Снижение функциональной активности клеток Купфера.

3.2.1.1.1. Численность КК и их функциональная активность.

3.2.1.1.2. Механизмы и стадийность депрессии функций КК.

3.2.1.2. Повышение функциональной ak i ивности Мф внепеченочных локализаций.

3.2.1.2.1. Механизмы стимуляции внепеченочных Мф.

3.2.1.2.2. Роль системной эндотоксинемии.

глава 3.2:2. механизмы и роль снижения реактивности смф в хронизации воспалительного процесса в печени.

3.2.2.1. При ССь4-гепатофиброзе.

3.2.2.1.1. Реакция СМФ как целого.

3.2.2.1.2. Механизмы снижения реактивности СМФ при гепатофиброзе.

3.2.2.2. При вирусных гепатитах (ВГ) человека.

3.2.2.2.1. Механизмы снижения реактивности Мф при ВГ.

часть 3.3. реактивность мф при атерогенезе.

глава 3.3.1. эндотоксинемия и лпс-липопротеиновые комплексы как факторы атерогенеза и гипореактивности мф.

3.3.1.1. формирование ЛПС-ЛП комплексов.

3.3.1.1.1. Роль разных классов ЛП в формировании ЛПС-ЛП комплексов.

3.3.1.1.2. Механизм формирования комплексов.

3.3.1.1.3. ЛПС-индуцированный спектр ЛП крови.

3.3.1.2. проа1ерогенные эффекты комплексов.

3.3.1.2.1. Ингибирование ЛХАТ.

3.3.1.2.2. ЛПС-опосредованная модификация ЛПНП и изменение клиренса ЛПНП.

3.3.1.2.3. Механизмы ЛПС-модификации ЛПНП.

3.3.1.2.4. Механизмы аккумуляции ЛПС-ЛП комплексов в ткани аорты.

3.3.1.2.5. Механизмы аккумуляции ЛПС-ЛП комплексов в Мф.

3.3.1.2.6. Реальная значимость ЛПС-зависимых механизмов в атерогенезе.

3.3.1.2.7. Возможная роль ЛПС-ЛП комплексов в регуляции реактивности Мф атеросклеротической бляшки.

ГЛАВА 3.3.2. ОКСИСТЕРОЛЫ В ФИБРОСКЛЕРОТИЧЕСКОЙ ТРАНСФОРМАЦИИ

АТЕРОСКЛЕРОТИЧЕСКОЙ БЛЯШКИ.

ЧАСТЬ 3.4. РОЛЬ МЕВАЛОНАТНОГО БИОХИМИЧЕСКОГО ПУТИ В РЕГУЛЯЦИИ

РЕАКТИВНОСТИ МФ И ФИБРОГЕНЕЗЕ.

ГЛАВА 3.4.1. РОЛЬ МЕВАЛОНАТНОГО ПУТИ В РЕГУЛЯЦИИ РЕАКТИВНОСТИ МФ

И ФИБРОГЕНЕЗЕ: ЭФФЕКТЫ IN VIVO.

3.4.1.1. Ассоциация ХС/ОС - фиброз - гипорелкгивность Мф.

3.4.1.2. ХС И MEBAJIOHAT в РЕГУЛЯЦИИ УРОВНЯ ФИБРОГЕННОГО ОТВЕТА И РЕАКТИВНОСТИ МФ в МОДЕЛЯХ НЕФРОСКЛЕРОЗА.

3.4.1.2.1. Модель хронического рабдомиолиза.

3.4.1.2.2. Модель унилатерапьной обструкции мочеточника.

3.4.1.3. ХС в регуляции уровня фиброгенного ответа и реактивности мф при асеп гическом перитоните: модель спаечной болезни.

ГЛАВА 3.4.2. РОЛЬ МЕВАЛОНАТНОГО ПУТИ В РЕГУЛЯЦИИ РЕАКТИВНОСТИ МФ И

ПРОДУКЦИИ ТФР-р: ЭФФЕКТЫ IN VITRO.

3.4.2.1. гипореактивность мф при действии ХС, ОС и аторвастатина - особенности эффектов.

3.4.2.2. МФ ПРОДУКЦИЯ ТФР-Р ПРИ ДЕЙС ГВИИ ХС И АГОНИСТОВ ЯДЕРНЫХ РЕЦЕПТОРОВ: ВЛИЯНИЕ МЕВ.

3.4.2.3. мев-зависимые механизмы фиброгенеза и гипореактивности мф.

3.4.2.3.1. Особенности использованных моделей.

3.4.2.3.2. Возможные механизмы формирования гипореактивности мононуклеарных фагоцитов.

3.4.2.3.3. Возможные фиброгенные механизмы, обусловленные ингибированием мевалонатного пути.

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962. РОССИЙСКАЯ АКАДЕМИЯ МЕДИЦИНСКИХ НАУК СИБИРСКОЕ ОТДЕЛЕНИЕ ИНСТИТУТ ТЕРАПИИ1. На правах рукописи1. ШВАРЦ Яков Шмульевич05201150496

963. РЕАКТИВНОСТЬ МАКРОФАГАЛЬНОЙ СИСТЕМЫ ПРИ ФИБРОПЛАСТИЧЕСКИХ ПРОЦЕССАХ ВОСПАЛИТЕЛЬНОГО ГЕНЕЗА: ОЦЕНКА, МЕХАНИЗМЫ РЕГУЛЯЦИИ И ПАТОГЕНЕТИЧЕСКОЕ ЗНАЧЕНИЕ1403.03. Патологическая физиология

964. Диссертация на соискание ученой степени доктора медицинских наук

965. Научный консультант -доктор медицинских наук профессор М.И.Душкин1. Новосибирск 20101. ОГЛАВЛЕНИЕ1. ВВЕДЕНИЕ .7

966. РАЗДЕЛ 1. ОБЗОР ЛИТЕРАТУРЫ.21

967. ЧАСТЬ 1.1. ОБЩАЯ ХАРАКТЕРИСТИКА ФИБРОПРОЛИФЕРАТИВНЫХ ПРОЦЕССОВ.21

968. ГЛАВА 1.1.3. СОВРЕМЕННЫЕ ПРЕДСТАВЛЕНИЯ О РОЛИ МФ В РАЗВИТИИ/1. НЕФРОСКЛЕРОЗ А.56.11.3.3.1. Значение Мн-Мф инфильтрации.5711.3.3.2. Продуценты СТ при развитии нефросклероза.5911.3.3.3. Деградация СТ при развитии нефросклероза.60

969. ЧАСТЬ 1.2. СОВРЕМЕННЫЕ ПРЕДСТАВЛЕНИЯ О РЕАКТИВНОСТИ МФ И СМФ В.1. ЦЕЛОМ.63

970. ЧАСТЬ 1.3. ЛИПИДНЫЙ ОБМЕН ПРИ ВОСПАЛЕНИИ И ЕГО УЧАСТИЕ В РЕГУЛЯЦИИ1. РЕАКТИВНОСТИ МФ.80

971. ГЛАВА 1.3.1. ИЗМЕНЕНИЯ ЛИПИДНОГО ОБМЕНА ПРИ ВОСПАЛЕНИИ.8013.1.1. ТРИГЛИЦЕРИДЫ.8013.1.2. MEBAJIOI IAI НЫЙ ПУТЬ.8113.1.3. ХОЛЕСЛЕРИН.83

972. ГЛАВА 1.3.2. ОКСИСТЕРОЛЫ, LXR И РЕАКТИВНОСТЬ МФ.8413.2.1. РОЛЬ ОКСИСТЕРОЛОВ И LXR В ЛИПИДНОМ ОБМЕНЕ ПРИ ВОСПАЛЕНИИ.8413.2.2. ОКСИСТЕРОЛЫ В LXR-НЕЗАВИСИМОЙ РЕГУЛЯЦИИ.8513.2.3. ОКСИСТЕРОЛЫ И LXR В РЕГУЛЯЦИИ ФУНКЦИЙ МФ ПРИ ВОСПАЛЕНИИ.86

973. РАЗДЕЛ 2. СОБСТВЕННЫЕ РЕЗУЛЬТАТЫ.90

974. ЧАСТЬ 2.1. МАТЕРИАЛЫ И МЕТОДЫ ИССЛЕДОВАНИЯ.90

975. ЧАСТЬ 2.2. РЕАКТИВНОСТЬ МФ ПРИ'ХРОНИЧЕСКОМ ВОСПАЛЕНИИ И

976. ПОСТВОСПАЛИТЕЛЬНОМ ФИБРОЗЕ.108«

977. ГЛАВА 2.2.2. ГИПЕРЛИПИДЕМИЯ И ЭНДОТОКСИНЕМИЯ КАК ФАКТОРЫ

978. ГЛАВА 2.2.3. ХАРАКТЕР ГРАНУЛЕМАТОЗНОГО ВОСПАЛЕНИЯ, ВЫЗВАННОГО

979. Мф, полученных у мышей, находившихся на ХС диете.18922.4.2.2.3. Продукция ТФР-ß в культуре толерантных к эндотоксину перитонеальных Мф и влияние на нее мевалоната.190

980. РАЗДЕЛ 3. ОБСУЖДЕНИЕ РЕЗУЛЬТАТОВ.192

981. ЧАСТЬ 3.1. РЕАКТИВНОСТЬ МФ И УРОВЕНЬ ФИБРОГЕННОГО ОТВЕТА ПРИ ГРАНУЛЕМАТОЗНОМ ВОСПАЛИТЕЛЬНОМ ПРОЦЕССЕ У МЫШЕЙ РАЗНЫХ ЛИНИЙ".192

982. ГЛАВА 3.1.2. МЕЖЛИНЕЙНЫЕ РАЗЛИЧИЯ РЕАКТИВНОСТИ МФ И ВЫРАЖЕННОСТЬ

983. ЧАСТЬ 3.2. РЕАКТИВНОСТЬ МФ ПРИ ССЬ4-ИНДУЦИРОВАННОМ ГЕПАТОФИБРОЗЕ

984. И ЗАТЯЖНОМ ВИРУСНОМ ГЕПАТИТЕ ЧЕЛОВЕКА.216

985. ГЛАВА 3.2.1. МЕХАНИЗМЫ ФУНКЦИОНАЛЬНОЙ ПЕРЕСТРОЙКИ ПЕЧЕНОЧНОГО

986. ЧАСТЬ 3.3. РЕАКТИВНОСТЬ МФ ПРИ АТЕРОГЕНЕЗЕ.236

987. ГЛАВА 3.3.2. ОКСИСТЕРОЛЫ В ФИБРОСКЛЕРОТИЧЕСКОЙ ТРАНСФОРМАЦИИ

988. АТЕРОСКЛЕРОТИЧЕСКОЙ БЛЯШКИ.251

989. ЧАСТЬ 3.4. РОЛЬ МЕВАЛОНАТНОГО БИОХИМИЧЕСКОГО ПУТИ В РЕГУЛЯЦИИ

990. РЕАКТИВНОСТИ МФ И ФИБРОГЕНЕЗЕ.253

991. ГЛАВА 3.4.1. РОЛЬ МЕВАЛОНАТНОГО ПУТИ В РЕГУЛЯЦИИ РЕАКТИВНОСТИ МФ

992. ГЛАВА 3.4.2. РОЛЬ МЕВАЛОНАТНОГО ПУТИ В РЕГУЛЯЦИИ РЕАКТИВНОСТИ МФ И