中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2009年
5期
471-474
,共4页
孟莹%李旭%蔡绍曦%周高速
孟瑩%李旭%蔡紹晞%週高速
맹형%리욱%채소희%주고속
血管紧张素Ⅱ%核转录因子κB%肺泡巨噬细胞%急性肺损伤
血管緊張素Ⅱ%覈轉錄因子κB%肺泡巨噬細胞%急性肺損傷
혈관긴장소Ⅱ%핵전록인자κB%폐포거서세포%급성폐손상
Angiotensin Ⅱ%NF-κB%Alveolar macrophages%Acute lung injury
目的 阐明血管紧张素Ⅱ(AngⅡ)对肺泡巨噬细胞核转录因子κB(NF-κB)信号通路的影响.方法 收集正常或其他肺病患者正常肺叶组织肺泡灌洗液,分离、纯化、培养肺泡巨噬细胞.予10-6M AngⅡ刺激15,30,60,120 min.另外,予AT-1受体阻断剂irbesartan(10-6M)预处理肺泡巨噬细胞60 min后再予10-6 M AngⅡ刺激60 min.设置对照组.凝胶电泳移动抑制实验(EMSA)检测NFκB的结合活性.免疫蛋白质印迹检测胞浆内IκBα的表达.RT-PCR检测TNF-α和ICAM-1的表达.应用SPSS 13.0软件进行One-Way ANOVA方差分析,多重比较采用LSD法.以P<0.05为差异具有统计学意义.结果 AngⅡ刺激肺泡巨噬细胞15 min NF-xB结合活性增强,60 min达到峰值.AT-1受体阻断剂irbesartan可阻断NF-κB结合活性增强.与对照组(1.0)相比,AngⅡ处理组(0.29±0.11)ⅠκBα旺表达减弱,且差异具有统计学意义(P=0.013).与AngⅡ处理组相比,AngⅡ+irbesartan处理组ⅠκBα表达(0.83±0.12)则增强,且差异具有统计学意义(P=0.001).与对照组(0.42±0.099)相比,AngⅡ处理组TNF-α(1.13±0.17)表达增强,且差异具有统计学意义(P=0.001).与AngⅡ处理组相比,AngⅡ+irbesartan处理组(0.77±0.15)减弱,且差异具有统计学意义(P=0.02).与对照组ICAM-1表达(0.16±0.050)相比,AngⅡ处理组(0.55±0.08)增强且差异具有统计学意义(P=0.003).与AngⅡ处理组相比,AngⅡ+irbesartan处理组(0.32±0.07)减弱,且差异具有统计学意义(P=0.001).结论 Ang Ⅱ对肺泡巨噬细胞NF-κB通路有正向调节作用.
目的 闡明血管緊張素Ⅱ(AngⅡ)對肺泡巨噬細胞覈轉錄因子κB(NF-κB)信號通路的影響.方法 收集正常或其他肺病患者正常肺葉組織肺泡灌洗液,分離、純化、培養肺泡巨噬細胞.予10-6M AngⅡ刺激15,30,60,120 min.另外,予AT-1受體阻斷劑irbesartan(10-6M)預處理肺泡巨噬細胞60 min後再予10-6 M AngⅡ刺激60 min.設置對照組.凝膠電泳移動抑製實驗(EMSA)檢測NFκB的結閤活性.免疫蛋白質印跡檢測胞漿內IκBα的錶達.RT-PCR檢測TNF-α和ICAM-1的錶達.應用SPSS 13.0軟件進行One-Way ANOVA方差分析,多重比較採用LSD法.以P<0.05為差異具有統計學意義.結果 AngⅡ刺激肺泡巨噬細胞15 min NF-xB結閤活性增彊,60 min達到峰值.AT-1受體阻斷劑irbesartan可阻斷NF-κB結閤活性增彊.與對照組(1.0)相比,AngⅡ處理組(0.29±0.11)ⅠκBα旺錶達減弱,且差異具有統計學意義(P=0.013).與AngⅡ處理組相比,AngⅡ+irbesartan處理組ⅠκBα錶達(0.83±0.12)則增彊,且差異具有統計學意義(P=0.001).與對照組(0.42±0.099)相比,AngⅡ處理組TNF-α(1.13±0.17)錶達增彊,且差異具有統計學意義(P=0.001).與AngⅡ處理組相比,AngⅡ+irbesartan處理組(0.77±0.15)減弱,且差異具有統計學意義(P=0.02).與對照組ICAM-1錶達(0.16±0.050)相比,AngⅡ處理組(0.55±0.08)增彊且差異具有統計學意義(P=0.003).與AngⅡ處理組相比,AngⅡ+irbesartan處理組(0.32±0.07)減弱,且差異具有統計學意義(P=0.001).結論 Ang Ⅱ對肺泡巨噬細胞NF-κB通路有正嚮調節作用.
목적 천명혈관긴장소Ⅱ(AngⅡ)대폐포거서세포핵전록인자κB(NF-κB)신호통로적영향.방법 수집정상혹기타폐병환자정상폐협조직폐포관세액,분리、순화、배양폐포거서세포.여10-6M AngⅡ자격15,30,60,120 min.령외,여AT-1수체조단제irbesartan(10-6M)예처리폐포거서세포60 min후재여10-6 M AngⅡ자격60 min.설치대조조.응효전영이동억제실험(EMSA)검측NFκB적결합활성.면역단백질인적검측포장내IκBα적표체.RT-PCR검측TNF-α화ICAM-1적표체.응용SPSS 13.0연건진행One-Way ANOVA방차분석,다중비교채용LSD법.이P<0.05위차이구유통계학의의.결과 AngⅡ자격폐포거서세포15 min NF-xB결합활성증강,60 min체도봉치.AT-1수체조단제irbesartan가조단NF-κB결합활성증강.여대조조(1.0)상비,AngⅡ처리조(0.29±0.11)ⅠκBα왕표체감약,차차이구유통계학의의(P=0.013).여AngⅡ처리조상비,AngⅡ+irbesartan처리조ⅠκBα표체(0.83±0.12)칙증강,차차이구유통계학의의(P=0.001).여대조조(0.42±0.099)상비,AngⅡ처리조TNF-α(1.13±0.17)표체증강,차차이구유통계학의의(P=0.001).여AngⅡ처리조상비,AngⅡ+irbesartan처리조(0.77±0.15)감약,차차이구유통계학의의(P=0.02).여대조조ICAM-1표체(0.16±0.050)상비,AngⅡ처리조(0.55±0.08)증강차차이구유통계학의의(P=0.003).여AngⅡ처리조상비,AngⅡ+irbesartan처리조(0.32±0.07)감약,차차이구유통계학의의(P=0.001).결론 Ang Ⅱ대폐포거서세포NF-κB통로유정향조절작용.
Objective To determine the effects of angiotensin Ⅱ (Ang Ⅱ) on NF-κB DNA binding activity in alveolar macrophage. Method Human alveolar macrophages were isolated and made homogeneous from alveo-lar lavage fluid, and cuhtured in DMEM. Alvcolar macrophages were treated with AugⅡ (10-6M) for 15 min, 30 min, 60 min and 120 min, respectively. Moreover, alveolar macmphages were pretreated with irbesartan (AngⅡ type 1 receptor blocker) for Ⅰ hour before stimulated with Angiotensin Ⅱ for Ⅰ hour. Electrophoretic gel mobility shift assay (EMSA) was used to detect NF-κB DNA binding activity. The protein expression of IκBα was examined by Western blot. Expressions of TNF-α and ICAM-1 mRNA were detected by using RT-PCR. Results EMSA re-vealed that there was a increase in up-regulation of NF-κB DNA binding activity after alveolar macrophages were treated with Ang Ⅱ for 15 rain and peaked at 60 min. Irbesartan treatment reduced DNA binding activity. Com-pared with control group, the protein expression of IκBα decreased in Ang Ⅱ treatment group(0.29±0.11, P= 0.013), and Irbesartan treatment significantly increased protein expression of IκBα(0.83±0.12, P=0.001). The expressions of TNF-α and ICAM-1 mRNA were up-regulated by AngⅡ in comparison with the control group (TNF-α:1.13±0.17 vs. 0.42±0.099; ICAM-1 0.55±0.08 vs. 0.16±0.050, P=0.003). Irbesartan inhibited the expressions of TNF-α (0.77±0.15 vs 1.13±0.17, P=0.02; ICAM-1(0.32±0.07 vs 0.55±0.08, P =0.001). Conclusions Ang Ⅱ is capable to stimulate NF-κB signal pathway in alveolar macrophages.
