灯盏花素对大鼠脑缺血再灌注损伤相关CYP4A的作用机制

目的:研究灯盏花素对大鼠脑缺血再灌注损伤相关细胞色素(cytochrome,CYP)4A1,CYP4A3,CYP4A8的作用机制。方法:采用SD雄性大鼠193只,制备大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)模型,在脑缺血2 h,再灌注术后立即尾静脉注射给予灯盏花素注射剂(12,6,3 mg·kg~(-1)),每日1次,连续7 d,末次给药后,断头取右脑组织,用实时荧光定量PCR法检测脑组织内的CYP基因CYP4A1,CYP4A3,CYP4A8的表达情况,用酶联反应吸附测定(enzyme linked immunosorbent assay,ELISA)法检测大鼠内皮型一氧化氮合酶(endothelial nitric oxide synthase,e NOS)含量。结果:与假手术组比较,模型组大鼠脑损伤168 h后的死亡率、脑梗死体积、行为学评分显著升高(P0.01),大鼠脑组织CYP4A3,CYP4A8显著上调(P0.01),e NOS含量显著增加(P0.01)。与模型组比较,灯盏花素(12,6 mg·kg-1)组可降低的大鼠脑损伤168 h的死亡率、脑梗死体积(P0.05),降低24 h后行为学评分(P0.01),下调大鼠脑组织CYP4A3,CYP4A8表达和血清e NOS含量(P0.05,P0.01);灯盏花素(3 mg·kg~(-1))组对降低大鼠脑损伤168 h后死亡率和脑梗死体积不明显,可明显降低120 h时行为学评分(P0.05),显著下调CYP4A3,CYP4A8表达和脑组织e NOS含量(P0.01)。结论:大鼠脑缺血再灌注损伤会加重大鼠死亡率、脑梗死体积、行为学评分,上调脑组织CYP4A3,CYP4A8的表达和增加脑组织及血清e NOS的含量。灯盏花素可降低大鼠脑缺血再灌注损伤大鼠死亡率和行为学评分,可能是通过下调脑组织CYP4A3,CYP4A8的表达来改善大鼠脑缺血再灌注损伤。

Mechanism of Breviscapine on Cerebral Ischemia Reperfusion Injury of Rats by CYP4A Pathway

Objective: To study the mechanism of Breviscapine on cerebral ischemia reperfusion injury of rats by cytochrome (CYP) 4A1, CYP4A3, CYP4A8 pathways.Method: Totally 193 male SD rats were selected to establish the model of middle cerebral artery occlusion (MCAO). After cerebral ischemia for 2 h and reperfusion, they were injected with Breviscapine at the doses of 12, 6, 3 mg·kg^-1, qd, for 7 days. After the final medication, tissue fluids isolated from brain tissues were prepared. Genes expressions of CYP4A1, CYP4A3 and CYP4A8 in brain tissues of rats were examined by Real-time PCR assays. The content of endothelial nitric oxide synthase (eNOS) was examined by enzyme linked immunosorbent assay(ELISA).Result: Compared with shame group, model group showed significant increase in motility (P<0.01), volume of cerebral infarction and behavioral score (P<0.01) at 168 h, and up-regulation in the expressions of CYP4A3, CYP4A8 (P<0.01). The content of eNOS was increased in brain tissues and serum. Compared with model group, Bre 12 mg·kg^-1 group and Bre 6 mg·kg^-1 group showed significant reduction in motility (P<0.05), volume of cerebral infarction (P<0.01) at 168 h, behavioral score (P<0.01) after 24 h, and down-regulation in the expressions of CYP4A3, CYP4A8 (P<0.05, P<0.01) and the content of eNOS (P<0.01) in brain tissues and serum. Bre 3 mg·kg^-1 group showed significant reduction in the behavioral score (P<0.01) at 120 h and the content of eNOS (P<0.01), and down-regulation in the expression of CYP4A3, CYP4A8 (P<0.01) in brain tissues.Conclusion: Motility, volume of cerebral infarction and behavioral score were increased by cerebral ischemia reperfusion injury, the expression of CYP4A3, CYP4A8 were up-regulated, and the content of eNOS was increased in brain tissues. Breviscapine injection could cure cerebral ischemia reperfusion injury of rats by reducing the volume of cerebral infarction, behavioral score, which may be correlated with down-regulation in the expressions of CYP4A3, CYP4A8 in brain tissues.