中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2008年
6期
606-609
,共4页
桂平%武庆平%姚尚龙%舒化青
桂平%武慶平%姚尚龍%舒化青
계평%무경평%요상룡%서화청
机械拉伸%γ-干扰素%肺泡上皮细胞%β防御素
機械拉伸%γ-榦擾素%肺泡上皮細胞%β防禦素
궤계랍신%γ-간우소%폐포상피세포%β방어소
Mechanical stretch%Interferon-gamma%Alveolar epithelial cell%Beta-defensins
目的 研究机械拉伸对γ-干扰素(IFN-γ)诱导肺泡上皮细胞(A549细胞)人β-防御素-3(HBD-3)表达的影响并探讨HBD-3在呼吸机相关性肺炎(VAP)发病机制中的作用.方法 体外培养A549细胞,分别给予机械拉伸(S组),10 ng/ml IFN-γ(I组)和10.ng/ml IFN-γ+机械拉伸(IS组)3种处理,未处理的细胞为对照组(C组),处理的时间为2 h、4 h和6 h.拉伸由细胞FlexerceH-4000TM拉伸系统提供,拉伸的幅度为20%,速率为30次/min.处理后实时定量逆转录-多聚酶链反应(RT-PCR)检测HBD-3 mRNA的表达;处理6 h的细胞继续培养至24 h,激光扫描共聚焦显微镜检测HBD-3的表达.采用单因素方差分析和q检验对实验数据进行统计学分析.结果 单独的机械拉伸不能使HBD-3mRNA的表达发生显著变化.10 ng/ml IFN-γ处理2 h、4 h和6 h后,HBD-3 mRNA表达量较之C组分别增加了(2.63±0.60),(8.02±1.63)和(12.21±2.35)倍.10 rig/ml IFN-γ+机械拉伸处理2 h、4 h和6 h后,HBD-3 mRNA表达量较之C组分别增加了(1.54±0.16),(4.37±0.87)和(6.99 4-1.32)倍,但均显著低于I组(P<0.01).6 h机械拉伸后,IS组HBD-3的表达显著少于I组(P<0.01),同C组差异无统计学意义.结论 机械拉伸能显著抑制IFN-γ诱导肺泡上皮细胞HBD-3表达的上调,这可能是机械通气(MV)的患者易于发生VAP的原因之一.
目的 研究機械拉伸對γ-榦擾素(IFN-γ)誘導肺泡上皮細胞(A549細胞)人β-防禦素-3(HBD-3)錶達的影響併探討HBD-3在呼吸機相關性肺炎(VAP)髮病機製中的作用.方法 體外培養A549細胞,分彆給予機械拉伸(S組),10 ng/ml IFN-γ(I組)和10.ng/ml IFN-γ+機械拉伸(IS組)3種處理,未處理的細胞為對照組(C組),處理的時間為2 h、4 h和6 h.拉伸由細胞FlexerceH-4000TM拉伸繫統提供,拉伸的幅度為20%,速率為30次/min.處理後實時定量逆轉錄-多聚酶鏈反應(RT-PCR)檢測HBD-3 mRNA的錶達;處理6 h的細胞繼續培養至24 h,激光掃描共聚焦顯微鏡檢測HBD-3的錶達.採用單因素方差分析和q檢驗對實驗數據進行統計學分析.結果 單獨的機械拉伸不能使HBD-3mRNA的錶達髮生顯著變化.10 ng/ml IFN-γ處理2 h、4 h和6 h後,HBD-3 mRNA錶達量較之C組分彆增加瞭(2.63±0.60),(8.02±1.63)和(12.21±2.35)倍.10 rig/ml IFN-γ+機械拉伸處理2 h、4 h和6 h後,HBD-3 mRNA錶達量較之C組分彆增加瞭(1.54±0.16),(4.37±0.87)和(6.99 4-1.32)倍,但均顯著低于I組(P<0.01).6 h機械拉伸後,IS組HBD-3的錶達顯著少于I組(P<0.01),同C組差異無統計學意義.結論 機械拉伸能顯著抑製IFN-γ誘導肺泡上皮細胞HBD-3錶達的上調,這可能是機械通氣(MV)的患者易于髮生VAP的原因之一.
목적 연구궤계랍신대γ-간우소(IFN-γ)유도폐포상피세포(A549세포)인β-방어소-3(HBD-3)표체적영향병탐토HBD-3재호흡궤상관성폐염(VAP)발병궤제중적작용.방법 체외배양A549세포,분별급여궤계랍신(S조),10 ng/ml IFN-γ(I조)화10.ng/ml IFN-γ+궤계랍신(IS조)3충처리,미처리적세포위대조조(C조),처리적시간위2 h、4 h화6 h.랍신유세포FlexerceH-4000TM랍신계통제공,랍신적폭도위20%,속솔위30차/min.처리후실시정량역전록-다취매련반응(RT-PCR)검측HBD-3 mRNA적표체;처리6 h적세포계속배양지24 h,격광소묘공취초현미경검측HBD-3적표체.채용단인소방차분석화q검험대실험수거진행통계학분석.결과 단독적궤계랍신불능사HBD-3mRNA적표체발생현저변화.10 ng/ml IFN-γ처리2 h、4 h화6 h후,HBD-3 mRNA표체량교지C조분별증가료(2.63±0.60),(8.02±1.63)화(12.21±2.35)배.10 rig/ml IFN-γ+궤계랍신처리2 h、4 h화6 h후,HBD-3 mRNA표체량교지C조분별증가료(1.54±0.16),(4.37±0.87)화(6.99 4-1.32)배,단균현저저우I조(P<0.01).6 h궤계랍신후,IS조HBD-3적표체현저소우I조(P<0.01),동C조차이무통계학의의.결론 궤계랍신능현저억제IFN-γ유도폐포상피세포HBD-3표체적상조,저가능시궤계통기(MV)적환자역우발생VAP적원인지일.
Objective To study the effect of mechanical stretch on the expression of human beta-defensin-3 (HBD-3) in alveolar epithelial cells(A549 cells) elicited by interferon-gamma(IFN-γ) and to investigate the role of HBD-3 in the pathogenesis of ventilator-associated pneumonia (VAP) . Method A549 cells cultured in vitro were treated with mechanical stretch (group S), 10 ng/ml IFN-γ (group I) ,and 10 ng/ml IFN-γ with mechanical stretch (group IS), respectively. Cells without treatment served as controls (group C). Cells were stretched by 20% amplitude of stretch at 30 cycles/mm by Flexercell-4000[TM] Unit for 2 h, 4 h, and 6 hours. The HBD-3 mRNA expression was determined by real-time RT-PCR after treatment. After 6 hours, treatment, cells were cultured for 24 hours and the expression of HBD-3 was examined by laser scanning confocal microscope. The experimental data were statistically analyzed by using one-way ANOVA analysis and q-test. Results The expression of HBD-3 mRNA in A549 cells could not significantly be changed by mechanical stretch alone. Compared with group C,the HBD-3 mRNA expression after treatment with 10 ng/ml IFN-γ for 2 hours,4 hours and 6 hours increased significantly by (2.63 C,the HBD-3 mRNA expression after treatment with 10 ng/ml IFN-γ and mechanical stretch for 2 hours,4 hours and 6 hours increased by (1.54 were significantly lower than those in group I (P < 0.01). The HBD-3 expression in group IS after mechanical stretch for 6 significantly different from than in group C. Conclusions Mechanical stretch can significantly suppress the up-regulation of HBD-3 in alveolar epithelial cells elicited by IFN-γ, and this may be one of the explaina-tions that patients under mechanical ventilatiori(MV) have a higher risk of VAP.
