摘要
目的探讨周期性机械牵张力对人支气管上皮细胞(16HBE)信号转导子与转录激活子3(STAT3)表达的作用效应。方法利用周期性机械牵张加力装置对16HBE细胞进行不同作用力、不同作用时间刺激,采用RT-PCR检测STAT3 mRNA的表达,Western blot测定STAT3蛋白的表达及磷酸化水平变化。结果不同机械牵张力刺激16HBE细胞30min时,3kPa作用力对STAT3酪氨酸705残基(Tyr705)磷酸化程度最高。在3kPa的周期性机械牵张刺激下,STAT3的酪氨酸残基的活化具有时间依赖性,在30min时与对照相比差异有统计学意义(P<0.05);1h时磷酸化程度最高(P<0.01);机械牵张力不影响丝氨酸727(Ser727)残基的磷酸化、STAT3蛋白表达量以及STAT3基因转录水平。结论STAT3通过Tyr705磷酸化修饰参与支气管上皮细胞对周期性机械牵张刺激的应答,在支气管上皮细胞对机械信号的应答和转导中起着重要作用。
Objective To study the effects of cyclic mechanical stretch on the activity and expression of STAT3 in Human bronchial epithelial Ceil （16HBEC）. Methods The 16HBECs were subjected to cyclic mechanical stretch by the four-point bending system at 0.5 Hz of 1 kPa, 2 kPa, 3 kPa, 4 kPa for 30 min or for 30 min, 1 h, 3 h, and 6 h of 3 kPa respectively. The mRNA level of STAT3 was semiuantified by RT-PCR; STAT3 protein and phosphorylated STAT3 were detected by western blot. Results Phosphorylation level of STAT3 Tyr705 was higher under 3 kPa cyclic mechanical stretch than the others. 3 kPa stretch phosphorylatd STAT3 Tyr705 in 16HBE cells in a timeependent manner. The phosphorylation of STAT3-TyrT05 was significantly increased at 30rain compared with the control group（P〈20.05）, and reached the highest phosphorylation level at lh, then showed a sharp descending tendency. There was no effect of 3 kPa stretch on the phosphorylation of STAT3 Ser727, expression of STAT3 protein or mRNA levels. Conclusion STAT3 played an important role in the response and signal transduetion of 16HBECs to the cyclic mechanical stretch stimulation through the phosphorylation of STAT3 Tyr705.
出处
《四川大学学报（医学版）》
CAS
CSCD
北大核心
2009年第4期672-675,共4页
Journal of Sichuan University(Medical Sciences)
基金
国家自然科学基金(批准号30425007
30370627
30670921)
纽约中华医学基金会(CMB
00-722
06-834)资助
关键词
周期性机械牵张
信号转导子与转录激活子3(STAT3)
人支气管上皮细胞
机械通气
Cyclic mechanical stretch Single transductors and activators of transcription 3 （STAT3） Human bronchial epithelial Cell（16HBEC） Mechanical ventilation
