法舒地尔对大鼠脑缺血再灌注损伤炎症反应的影响

目的：观察法舒地尔对大鼠脑缺血再灌注损伤炎症反应的影响,探讨其抗炎机制。方法：大脑中动脉线栓法（MCAO）制作大鼠局灶性脑缺血再灌注损伤模型,缺血1.5h再灌注24h。法舒地尔术前腹腔注射给药15mg/kg,术后12h再次给药。术后对大鼠神经功能进行评分,TTC染色观察脑梗死体积;用干湿重法测定脑含水量;分光光度法测定髓过氧化物酶（MPO）活性;伊文思兰法（EB）测定血脑屏障的损伤程度;免疫组化检测大鼠脑缺血区细胞间黏附分子-1（ICAM-1）、血管细胞黏附分子-1（VCAM-1）、NF-κB p65的表达;ELISA法检测IL-8的含量;Western blot法检测单核细胞趋化蛋白-1（MCP-1）和核抽提物中NF-κBp65蛋白的表达;RT-PCR检测NF-κB p65mRNA的表达。结果：法舒地尔能明显改善脑缺血再灌注损伤大鼠神经缺陷症状,缩小脑梗死体积,明显降低缺血侧脑组织的含水量、EB含量及MPO活性;显著抑制ICAM-1、VCAM-1、IL-8和MCP-1蛋白的表达;降低NF-κB p65mRNA和蛋白的表达;减少脑组织核抽提物中NF-κB p65的蛋白量（P〈0.05vsMCAO组）。结论：法舒地尔通过抑制NF-κBp65的活化,进而抑制黏附分子及趋化因子的表达,减轻脑缺血再灌注损伤的炎症反应。

Effects of fasudil on inflammatory reaction in cerebral ischemia/reperfusion rats

AIM： To observe the effects of fasudil on inflammatory reaction in cerebral ischemia/reperfusion rats. METHODS： Focal cerebral ischemia/reperfusion model in rats was made by transient occlusion of middle cerebral artery for 1.5 h followed by 24 h reperfusion. Fasudil was injected intraperitoneally before operation and 12 h after operation. The neurological function was scored and the infarct volume was measured after operation. The water content of brain was measured with dry-wet weight; The myeloperoxidase （MPO） activity was determined by spectrophotometer; The permeability of the blood brain barrier was evaluated by measurement of the Evans Blue （EB） content in the brain with spectrophotometer; The brain content of IL-8 was evaluated with ELISA; The level of NF-κB p65 mRNA expression was determined by RT- PCR; The expressions of ICAM-1, VCAM-1 and NF-κB p65 protein were observed by immunohistochemistry; The MCP-1 and the expression of NF-κB p65 protein in the nuclear extracts were detected by western blotting. RESULTS： Fasudil improved the neurological function, decreased the infarct size, reduced the permeability of blood brain barrier and brain water content, inhibited the MPO activity, decreased the content of IL-8 and the protein expressions of ICAM-1, VCAM- 1 and MCP-1 in brain tissue, inhibited the expressions of NF-κB p65 mRNA and protein and reduced the content of NF-κB p65 protein in the nuclear extracts significandy（P 〈0.05 vs Model Group）. CONCLUSION： These results show that the anti-inflammatory effect of fasudil may be correlated with decreasing the expression of adhesion molecule and chemotactic factors though inhibiting the activity of NF-κB. KEY WORDS