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瞬时感受器电位通道在人脑微血管内皮细胞机械牵张中的作用及其信号转导机制

, PP. 2183-2186

Keywords: 机械牵张刺激,瞬时感受器电位通道,钙离子内流

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Abstract:

目的观察瞬时感受器电位(transientreceptorpotential,TRP)通道在人脑微血管内皮细胞(humanbrainmicrovascularendothelialcells,HBMEC)机械牵张刺激中的作用,并对其信号转导入胞内的机制进行初步探讨。方法HBMEC体外培养,采用多功能小型生物撞击机给予机械牵张刺激,根据不同施加条件将细胞分为对照、单纯牵张、牵张+SKF-96365(非选择性TRP通道阻滞剂)、牵张+低分子量肝素(lowmolecularweightheparin,LMWH)、牵张+一氧化氮合酶抑制剂(N-nitro-l-argininemethylester,L-NAME)共5组。采用激光扫描共聚焦显微镜观察胞内游离Ca2+分布情况,流式细胞仪检测牵张前后胞内Ca2+荧光强度变化;Westernblot检测牵张前后一氧化氮合酶(endothelialnitricoxidesynthase,eNOS)蛋白表达情况。结果流式细胞仪检测结果显示,单纯牵张组和牵张+LMWH组细胞内游离Ca2+浓度较对照组显著升高(P<0.05);牵张+SKF-96365组胞内Ca2+浓度较单纯牵张组明显降低(P<0.05)。Westernblot检测结果显示,单纯牵张组细胞eNOS蛋白表达水平明显增高(P<0.05);牵张+SKF-96365组和牵张+L-NAME组eNOS蛋白表达水平较单纯牵张组明显降低(P<0.05)。结论HBMEC胞膜TRP通道可感知机械牵张刺激的激活,继而引起胞内游离钙离子浓度升高,其转导机制可能主要与胞外Ca2+内流相关。

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