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目的:探讨粉防己碱对脑缺血/再灌注(I/R)后中性粒细胞募集反应的作用及分子机制。方法:大鼠()脑中动脉线栓闭塞(2h)/再通法建立局灶性脑缺血/再灌注模型。采用干湿重法、~(51)Cr标记中性粒细胞法、逆转录聚合酶链反应和凝胶迁移电泳分别检测脑组织含水量、中性粒细胞募集、细胞间粘附分子-1(ICAM-1)mRNA转录的水平以及细胞核内激活的核因子-кB(NF-кB)的水平。结果:脑含水量和中性粒细胞募集数量从再灌注3h到24h平行增加。在再灌注1h可检测到ICAM-1 mRNA的表达,12h达到最高值,24h下降到3h的水平。在再灌注0.5h可检测到NF-кB的激活,6h达到最高值,24h下降到1h的水平。在再灌注6h、12h和24h,粉防己碱(10和20mg/kg)可抑制这些变化。结论:粉防己碱可抑制脑缺血/再灌注后中性粒细胞的募集,ICAM-1 mRNA的转录和NF-кB的激活。
Objective: To investigate the effect and molecular mechanism of tetrandrine on neutrophil recruitment after cerebral ischemia/reperfusion (I/R). METHODS: A rat model of middle cerebral artery occlusion (2h)/recanalization was used to establish a model of focal cerebral ischemia/reperfusion. The brain tissue water content, neutrophil recruitment, and intercellular adhesion molecule-1 were measured by dry-wet weight method, ~(51)Cr labeled neutrophil method, reverse transcription polymerase chain reaction, and gel migration electrophoresis. The level of ICAM-1) mRNA transcription and the activation of nuclear factor-kappaB (NF-kappaB) levels in the nucleus. RESULTS: Brain water content and neutrophil recruitment increased in parallel from 3h to 24h after reperfusion. The expression of ICAM-1 mRNA was detected at 1h after reperfusion, reached the highest value at 12h, and decreased to 3h at 24h. The activation of NF-кB was detected at 0.5 h after reperfusion, reached the highest value at 6 h, and decreased to 1 h at 24 h. Tetrandrine (10 and 20 mg/kg) inhibited these changes at 6 h, 12 h, and 24 h after reperfusion. CONCLUSION: Tetrandrine can inhibit the recruitment of neutrophils, the transcription of ICAM-1 mRNA and the activation of NF-кB after cerebral ischemia/reperfusion.