5-LOX-CysLTs-CysLTsR表达规律及黄芩苷-栀子苷配伍对大鼠脑缺血抗炎作用机制研究

Study on the Expression Pattern of 5-LOX-CysLTs-CysLTsR and the Anti-inflammatory Mechanism of Baicalin-Germanuin on Cerebral Ischemia in Rats

目的动态观察脑卒中大鼠脑组织半胱氨酰白三烯(CysLTs)活性及其5-脂氧酶(5-LOX)、半胱氨酰白三烯受体1(CysLTs1)、半胱氨酰白三烯受体2(CysLTs2)表达水平变化规律,探讨黄芩苷-栀子苷(7:3)配伍抗脑缺血炎性损伤机制.方法线栓法制备大脑中动脉堵塞模拟脑缺血模型,模型成功后随机分为空白对照组(Control)、模型组(pMCAO)、黄芩苷-栀子苷(7:3)30mg/kg、45mg/kg、60mg/kg组、齐留通45mg/kg组、孟鲁司特0.5mg/kg组,持续35d给药治疗,每隔7d,神经功能评分观察神经保护作用,酶联免疫吸附实验(ELISA)法测脑组织CysLTs含量;蛋白质印迹(WesternBlot)法检测脑组织5-LOX、CysLTs1、CysLTs2蛋白表达水平.结果与模型组相比,黄芩苷-栀子苷(7:3)30mg/kg、45mg/kg、60mg/kg组在脑缺血28d内可显著降低了CysLTs水平;WesternBlot结果显示,5-LOX、CysLTs1和CysLTs2蛋白表达明显降低;至恢复期35d,各蛋白表达水平已无显著性差异.结论黄芩苷-栀子苷(7:3)配伍可减轻MCAO大鼠的神经功能障碍,其作用机制可能与抑制5-LOX活性、降低CysLTs1和CysLTs2蛋白表达有关.

Objective To dynamicly observe cysteinyl leukotriene(CysLTs) activity in brain tissue of stroke rats and its 5-lipoxygenase(5-LOX), cysteinyl leukotriene receptor 1(CysLTs1), cysteinyl The change of expression level of leukotriene receptor 2(CysLTs2), and to investigate the mechanism of anti-cerebral ischemic injury induced by baicalin-germangoside(7:3). Methods The model of middle cerebral artery occlusion simulated cerebral ischemia was prepared by suture embolism.The model was then randomly divided into blank control group(control), model group(pMCAO), baicalin-geniposide(7:3) 30 mg/kg, 45 mg/kg, 60 mg/kg groups, ziliutong 45 mg/kg group and montelukast 0.5 mg/kg group. The treatment lasted 35 days. Neuroprotective effect was observed by neurological function score every 7 days. The content of CysLTs in brain tissue was measured by ELISA, and the expression levels of 5-LOX, CysLTs1 and CysLTs2 in brain tissue were detected by Western Blot.Results Compared with the model group, baicalin-geniposide(7:3) 30 mg/kg, 45 mg/kg and 60 mg/kg groups could significantly reduce the level of CysLTs within 28 days after cerebral ischemia;Western Blot results showed that the expression of 5-LOX, CysLTs1 and CysLTs2 protein was significantly decreased;35 days after recovery, there was no significant difference in the expression levels of each protein. Conclusion Baicalin-geniposide(7:3) combination can alleviate neurological dysfunction in MCAO rats, and its mechanism may be related to inhibiting 5-LOX activity, reducing the expression of CysLTs1 and CysLTs2 protein.