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15mgkg,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-κBp65的表达;ELISA法检测IL-8的含量;Westernblot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-κBp65mRNA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.L8和MCP-1蛋白的表达;降低NF-κBp65mRNA和蛋白的表达;减少脑组织核抽提物中NF+κBp65的蛋白量(K0.05vsMCAO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 /reper-fusion rats

AIM: To observe the effects of fasudil on inflammatory reaction in cerebral ischemia /reperfu-sion rats. METHODS: Focal cerebral ischemia /reper-fusion model in rats was made by transient occ lusion of middle cerebral arteryy 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 spectropho tometer;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 L-8 was ev aluated with ELISA; The level of NF+κB p65 mRNA expression was detemined by RT-PCR; The expressions of ICAM-1，VCAM-1 and NF-κB p65 protein were observed by immunohistoche-mistry; The MCP-1 and the expression of NF-κB p65plotein in the nuc lear extracts were detected by westernblotting. RESULTS: Fasudil improved the neurologi-cal function, decreased the infarct size,reduced the permeability of blood brain barrier and brain water content, inhibited the MPO activity, decreased the ontent of lL-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 ex tracts signifi cantly(P< 0.05 vs Model Group). CONCLUSION: These results show that the antinflammatory effect of fasudil may be correlated w ith decreasing the expression of adhesion molecu le and chemotactic factors though in hibiting the activity of NF-κB.