Arginine stimulates intestinal cell migration through a focal adhesion kinase dependent mechanism

Abstract

Background: L-Arginine is a nutritional supplement that may be useful for promoting intestinal repair. Arginine is metabolised by the oxidative deiminase pathway to form nitric oxide (NO) and by the arginase pathway to yield ornithine and polyamines. Aims: To determine if arginine stimulates restitution via activation of NO synthesis and/or polyamine synthesis. Methods: We determined the effects of arginine on cultured intestinal cell migration, NO production, polyamine levels, and activation of focal adhesion kinase, a key mediator of cell migration. Results: Arginine increased the rate of cell migration in a dose dependent biphasic manner, and was additive with bovine serum concentrate (BSC). Arginine and an NO donor activated focal adhesion kinase (a tyrosine kinase which localises to cell matrix contacts and mediates b1 integrin signalling) after wounding. Arginine stimulated cell migration was dependent on focal adhesion kinase (FAK) signalling, as demonstrated using adenovirus mediated transfection with a kinase negative mutant of FAK. Arginine stimulated migration was dependent on NO production and was blocked by NO synthase inhibitors. Arginine dependent migration required synthesis of polyamines but elevating extracellular arginine concentration above 0.4 mM did not enhance cellular polyamine levels. Conclusions: These results showed that L-arginine stimulates cell migration through NO and FAK dependent pathways and that combination therapy with arginine and BSC may enhance intestinal restitution via separate and convergent pathways.

Effect of Arginine Deiminase From Streptococcus Pyogenes on Cytoskeleton Structure and Migration Activity of Human Endothelial Cells

“…В экспериментах с эндотелиальными клетками линии TR-ВВВ индуцированная под влиянием провоспалитель-ных цитокинов iNOS-зависимая продукция NO полностью ингибировалась после воздействия АД и полностью восстанавливалась при добавле-нии в культуральную среду экзогенного аргинина [5]. Показано, что миграция клеток эпителия ки-шечника связана с аргинин-зависимой продук-цией клетками NO, активностью FAK -киназы фокальных контактов [17] и киназой pp70s6k -ключевым регулятором 5' -концевой олигопи-римидин mRNA трансляции [18]. Полиамины синтезируются во всех типах клеток и участвуют в процессах транскрипции, трансляции и репли-кации [6].…”

“…Полиамины синтезируются во всех типах клеток и участвуют в процессах транскрипции, трансляции и репли-кации [6]. От синтеза полиаминов зависит мигра-ция клеток кишечного эпителия [17], а снижение уровня синтеза полиаминов подавляет формиро-вание ламеллиподий и стресс-волокон в мигри-рующих клетках [13,21].…”

ROLE OF mTOR SIGNALING IN INTESTINAL CELL MIGRATION

“…Previous work (29) in piglet and rat enterocytes showed that ARG and bovine serum concentrate were the best stimulators of cell migration. We also found in those studies that cycloheximide blocked this effect, implicating mRNA translation in this process (M. Rhoads and W. Chen, unpublished observations).…”

“…We used intestinal monolayer wounding with a razor blade, as described by McCormack et al (23) to determine whether amino acids stimulate intestinal cell migration, as previously described (29). Cdx2 cells were plated in six-well plates.…”

“…1A). In this and subsequent experiments, ARG was added at 4 mM, which was previously found to be the optimal concentration to stimulate migration (29). Rapidly migrating ARG-or BSC-treated cells were pretreated for 30 min with inhibitors.…”