Nan fang yi ke da xue xue bao = Journal of Southern Medical University · 2007
[Signal transduction in respiratory syncytial virus infection-induced thymic stromal lymphopoietin expression in human epithelial cells].
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод пока не подготовлен
Обработка статьи · этапы, ошибки и управление
Загрузка состояния…
Ошибки и ограничения
История · было → стало
Состояние обновляется каждые 5 секунд, пока блок открыт. Процент показывается только там, где известен объём работы. Пауза останавливает продолжение этапа; текущий запрос к источнику может завершиться.
Tu Hai-Yan, Chen Xin, Li Jing
Аннотация
OBJECTIVE: To explore the mechanism of signal transduction in respiratory syncytial virus (RSV)-induced expression of thymic stromal lymphopoietin (TSLP) in bronchial epithelial cells.
METHODS: The eukaryotic expression vector of RSV F protein, pcDNA3-F, was constructed and transfected into in vitro cultured human bronchial epithelial cell line CRL-9483, which was also transfected with Smad7 expression vector pcDNA3/Smad7 or exposed to p38, ERK1/2, JNK, and JAK/STAT1 inhibitors. The mRNA levels of TSLP and the housekeeping GAPDH gene were analyzed 24 h later with semi-quantitative RT-PCR. In cells with downregulated TSLP mRNA expression due to the addition of the signal inhibitors, cytoplasm TSLP or GAPDH protein levels were further analyzed using Western blotting.
RESULTS: Virtually no TSLP mRNA expression was detected by RT-PCR in cultured CRL-9483 cells transfected with pcDNA3 exclusively (TSLP/GAPDH relative total gray scale of 0.10-/+0.03), while cell transfection with pcDNA3-F resulted in significantly increased TSLP mRNA level (0.42-/+0.20, P=0.024). In the presence of F protein expression, both p38 and JNK inhibitors could downregulate TSLP mRNA levels (0.14-/+0.04, P=0.036; 0.23-/+0.07, P=0.048, respectively), while TGF-beta-Smad inhibiting protein Smad7 (0.60-/+0.25), ERK 1/2 inhibitor (0.45-/+0.23), and JAK/STAT1inhibitor (0.44-/+0.25) failed to block TSLP expression (P>0.05). Western blotting showed that p38 inhibitor (TSLP/GAPDH relative total gray scale=3.67-/+1.18, P=0.018) and JNK inhibitor (1.48-/+0.77, P=0.004) also downregulated the protein levels of TSLP as compared with pcDNA3-F transfection group (8.13-/+2.20).
CONCLUSION: RSV F protein can stimulate TSLP expression in human bronchial epithelial cells mediated partially by p38 and JNK signal pathways.
METHODS: The eukaryotic expression vector of RSV F protein, pcDNA3-F, was constructed and transfected into in vitro cultured human bronchial epithelial cell line CRL-9483, which was also transfected with Smad7 expression vector pcDNA3/Smad7 or exposed to p38, ERK1/2, JNK, and JAK/STAT1 inhibitors. The mRNA levels of TSLP and the housekeeping GAPDH gene were analyzed 24 h later with semi-quantitative RT-PCR. In cells with downregulated TSLP mRNA expression due to the addition of the signal inhibitors, cytoplasm TSLP or GAPDH protein levels were further analyzed using Western blotting.
RESULTS: Virtually no TSLP mRNA expression was detected by RT-PCR in cultured CRL-9483 cells transfected with pcDNA3 exclusively (TSLP/GAPDH relative total gray scale of 0.10-/+0.03), while cell transfection with pcDNA3-F resulted in significantly increased TSLP mRNA level (0.42-/+0.20, P=0.024). In the presence of F protein expression, both p38 and JNK inhibitors could downregulate TSLP mRNA levels (0.14-/+0.04, P=0.036; 0.23-/+0.07, P=0.048, respectively), while TGF-beta-Smad inhibiting protein Smad7 (0.60-/+0.25), ERK 1/2 inhibitor (0.45-/+0.23), and JAK/STAT1inhibitor (0.44-/+0.25) failed to block TSLP expression (P>0.05). Western blotting showed that p38 inhibitor (TSLP/GAPDH relative total gray scale=3.67-/+1.18, P=0.018) and JNK inhibitor (1.48-/+0.77, P=0.004) also downregulated the protein levels of TSLP as compared with pcDNA3-F transfection group (8.13-/+2.20).
CONCLUSION: RSV F protein can stimulate TSLP expression in human bronchial epithelial cells mediated partially by p38 and JNK signal pathways.