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Выделение и культивирование миофибробластов печени крыс методом эксплантации

Выделение и культивирование миофибробластов печени крыс методом эксплантации

Abstract

При развитии фиброза печени источником соединительной ткани являются миофибробласты, происходящие из двух популяций клеток печени: звёздчатых клеток печени и портальных фибробластов. Маркёром миофибробластов является экспрессия ƒ-гладкомышечного актина (ƒ-SMA). Отличительным признаком миофибробластов, происходящих из звёздчатых клеток печени, является сохранение экспрессии маркёра звёздчатых клеток десмина. Процесс активации, пролиферации и трансдифференцировки клеток в миофибробласты находится в тесной зависимости от активности транскрипционного фактора NF-kB и его ингибитора IkBƒ. Целью нашей работы было получение культуры миофибробластов печени, изучение их происхождения, фенотипа, связи экспрессии NF-kB и IkBƒ с процессами активации и трансдифференцировки звёздчатых клеток печени в миофибробласты. Для этого методом эксплантации мы выделили гетерогенную популяцию клеток печени крысы. Практически все полученные нами клетки экспрессировали десмин и ƒ-SMA. На основании этого мы предполагаем, что полученная нами культура миофибробластов являлась производной звёздчатых клеток печени, а единичные десминотрицательные клетки производными портальных фибробластов. Таким образом, звёздчатые клетки печени обладают большим потенциалом к активации, росту, пролиферации и трансдифференцировке в миофибробласты по сравнению с портальными фибробластами. Подтверждением активированного состояния клеток явилась устойчивая экспрессия NF-kB и его ингибитора IkBƒ во всех клетках на протяжении всего эксперимента.

During liver fibrosis development connective tissue is produced by myofibroblasts that could originate from two hepatic populations: hepatic stellate cells and portal fibroblasts. A marker of myofibroblasts is the expression of ƒ-smooth muscle actin (ƒ-SMA). Distinctive feature of myofibroblasts, derived from hepatic stellate cells, is the preservation of the hepatic stellate cells marker expression desmin. The processes of activation, proliferation and cells trans-differentiation into myofibroblasts are closely related to the activity of transcription factor NF-kB and its inhibitor IkBƒ. The aim of our work was to obtain a culture of hepatic myofibrobasts, to study their origin, phenotype, relations between NF-kB and IkBƒ expression and the processes of activation and cells trans-differentiation into myofibroblasts. For this purpose we isolated heterogeneous population of cells from rat liver by the method of explantation. Almost all the cells had desmin and ƒ-SMA expression. On this basis, we suppose that these myofibroblasts were hepatic stellate cells derivatives, and singular desmin-negative cells originated from portal fibroblasts. Thus, hepatic stellate cells have major potential to activation, growth, proliferation and transdifferentiation into myofibroblasts in comparison to portal fibroblasts. Activated state of the cells was confirmed by stable expression of NF-kB and its inhibitor IkBƒ in all the cells throughout the whole experiment.

Keywords

ЗВЁЗДЧАТЫЕ КЛЕТКИ ПЕЧЕНИ, ПОРТАЛЬНЫЕ ФИБРОБЛАСТЫ, ТРАНСКРИПЦИОННЫЙ ФАКТОР NF-KB, ДЕСМИН, ƒ-ГЛАДКОМЫШЕЧНЫЙ АКТИН, ИНГИБИТОР IKBƒ, ƒ-SMA, INHIBITOR IKBƒ

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
Average
Average
Average