银杏二萜内酯对大鼠脑缺血再灌注后氧化应激和炎症反应抑制作用的研究

The Research of Diterpene Ginkgolides Meglumine Injection on Oxidative Stress and Inflammatory Reaction after Cerebral Ischemia-Reperfusion in Rat

目的研究银杏二萜内酯葡胺注射液(DGMI)对大鼠局灶性脑缺血再灌注(I/R)后氧化应激和炎症反应的抑制作用,并探讨其可能的作用机制。方法设假手术组、I/R组和DGMI低、中、高剂量组,各28只;采用改良线栓法复制局灶性脑I/R大鼠模型,术后第2天开始腹腔注射给药,疗程7 d。盲法对神经功能损伤进行评分,失重法计算脑组织含水量,TTC染色法计算脑组织梗死体积百分比;生化分析脑组织氧化酶活性和丙二醛(MDA)含量;测定脑组织中一氧化氮(NO)含量和诱导型一氧化氮合酶(inducible NOS,iNOS)活性;通过HE染色、TUNEL法分别进行脑组织病理学检查及神经元凋亡状况观察;Western blotting法检测核转录因子-κB(NF-κB)蛋白表达;酶联免疫法测定脑组织炎症细胞因子含量。结果经DGMI中、高剂量治疗7 d能显著降低I/R大鼠神经功能缺损评分,降低脑组织含水量和梗死体积百分比;提高脑组织中抗氧化酶超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性,并降低MDA含量;改善脑组织形态结构病变和神经元凋亡状况,降低凋亡指数(apoptosis Index,AI),下调NF-κB蛋白表达;降低炎症细胞因子白介素-1β(IL-1β)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)含量;与I/R组比较差异均具有统计学意义(P<0.05或P<0.01)。结论DGMI对脑I/R大鼠氧化应激损伤和炎症反应均具有一定的抑制作用。

Objective To investigate the effect and mechanism of Diterpene Ginkgolides Meglumine Injection(DGMI)on oxidative stress and inflammatory reaction after cerebral ischemia reperfusion(I/R)in rats.Methods The rats were divided into sham operation group,I/R group,and DGMI low,medium and high dose groups with 28 rats each group.The rat model of focal cerebral I/R was established with Zea-Longa occluding suture.The drugs were given by intraperitoneal injection from the second day after operation for 7 days.The neurological deficits,ratio of infarct volume,and water content of the brain were evaluated.The oxidase activity and malondialdehyde(MDA)content in brain tissue were analyzed.The nitric oxide(NO)content and inducible nitric oxide synthase(inducible NOS,iNOS)activity in brain tissue were measured.The morphological changes of brain tissue were observed by Hematoxylin-Eosin(HE)staining,the cell apoptosis were observed after TUNEL staining.The expression of nuclear factor kappa-B(NF-κB)were determined by western blotting.Results Compared with data in the I/R group,the neurological deficit score was reduced,water content and infarct volume percentage in the brain tissue were reduced,and the antioxidant enzymes superoxide dismutase(SOD)and catalase in brain tissue(CAT)activity increased,MDA content decreased,the morphological changes in the brain tissue and neuronal apoptosis improved,apoptosis index(AI)decreased,NF-κB protein expression was down-regulated;inflammatory cytokine interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)levels were reduced in medium-dose DGM group and high-dose DGM group after 7 days of treatment(P<0.05 or P<0.01).Conclusion DGMI has a certain inhibitory effect on oxidative stress injury and inflammatory response in brain tissue in I/R rats.