银杏内酯B对脑出血大鼠神经细胞凋亡的影响

目的 观察银杏内酯B对大鼠脑出血后神经细胞凋亡、血清超氧化物歧化酶活性及丙二醛、白介素-1β、肿瘤坏死因子-α和白介素-6质量浓度的影响,探讨银杏内酯B抗神经细胞凋亡的机制.方法 选用健康雄性Wistar大鼠175只,随机(随机数字法)分成假手术组、脑出血组、大剂量组、中剂量组及小剂量组,每组35只,右侧尾状核注射大鼠自体血制作基底节区脑出血动物模型,分别于脑出血模型形成后给予1 mL 0.9％氯化钠注射液及不同剂量银杏内酯B(5,10,20 mg/kg)腹腔注射,每日一次共5d,在脑出血后2,6,12h和1,2,3,5d断头取血和获取脑组织,利用黄嘌呤氧化酶法检测血清超氧化物歧化酶活性,利用硫代巴比妥酸法测定血清丙二醛浓度,利用酶联免疫吸附试验测定血清白介素-1β、肿瘤坏死因子-α和白介素-6浓度,利用原位末端转移酶标记技术检测脑组织中凋亡神经细胞,各参数多组间比较采用方差分析,两组间比较采用LSD法.结果 脑出血后2,6,12 h和1,2,3,5d,小剂量组大鼠凋亡神经细胞数量、血清超氧化物歧化酶活性及丙二醛、白介素-1β、肿瘤坏死因子-α和白介素-6浓度与脑出血组比较差异无统计学意义(P＞0.05)；脑出血后12 h和1,2,3,5d,大剂量和中剂量组大鼠凋亡神经细胞数量、血清超氧化物歧化酶活性及丙二醛、白介素-1β、肿瘤坏死因子-α和白介素-6浓度较脑出血组显著减少(P＜0.05),脑出血后2h和6h,大剂量和中剂量组大鼠凋亡神经细胞数量、血清超氧化物歧化酶活性及丙二醛、白介素-1β、肿瘤坏死因子-α和白介素-6浓度与脑出血组比较差异无统计学意义(P＞0.05).结论 银杏内酯B可能通过降低体内自由基和炎症反应,从而抑制脑出血后神经细胞凋亡,达到神经保护作用.

The experimental study of ginkgolide B effectual on neuronal cell apoptosis in the rat with intracerebral hemorrhage

Objective To investigate the effects of ginkgolide B on neuronal cell apoptosis,superoxide dismutase activity,malondialdehyde,interleukin-1beta,tumor necrosis factor-alpha,and interleukin-6 levels in serum of rats with intracerebral hemorrhage in order to explore the role of ginkgolide B in suppressing the neuronal cell apoptosis.Methods A total of 175 male Wistar rats were randomly (random number)divided into sham operation group,intracerebral hemorrhage group,as well as low,medium and high dose treatment groups.The rat model of intracerebral hemorrhage was made with infusion of autologous whole blood to caudate nucleus in the right basal ganglia region.Ginkgolide B in dose of 5 mg/kg,10 mg/kg and 20 mg/kg was given to rats in the low,middle and high dose treatment groups by intraperitoneal injection once a day for 5 days after intracerebral hemorrhage.The rats with intracerebral hemorrhage in the sham operation groups received intraperitoneal administration of 1 mL saline.Animals were sacrificed by decapitation 2,6,12,24,48,72 h and 5 days after intracerebral hemorrhage.Brains were taken and blood samples were collected.Neuronal cell apoptosis was measured by using terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling(TUNEL),and superoxide dismutase activity in serum was determined by using xanthine oxidase method,and serum malondialdehyde level was detected by using thiobarbituric acid reactive substance assay,and interleukin-1beta,tumor necrosis factor-alpha,and interleukin-6 levels in serum were assayed with enzyme linked immunosorbent assay(ELISA).Statistical analysis was carried out by using one-way analysis of variance and least-significant difference test.Results As 2 h,6 h,12 h,24 h,48 h,72 h,and 5 days after intracerebral hemorrhage,the differences in the number of apoptotic neuronal cell,superoxide dismutase activity in serum,serum malondialdehyde,interleukin-1 beta,tumor necrosis factor-alpha and interleukin-6 levels between the low dose treatment group and intracerebral hemorrhage group were not significant statistically(P ＞0.05).As 12 h,24 h,72 h,and 5 days after intracerebral hemorrhage,the number of apoptotic neuronal cell,superoxide dismutase activity in serum,serum malondialdehyde,interleukin-1 beta,tumor necrosis factor-alpha and interleukin-6 levels in the medium dose and high dose treatment groups were significantly statistically lower than those in the intracerebral hemorrhage group(P ＜ 0.05),but these differences in above biomarkers were not significant statistically among these three groups 2 and 6 hours after intracerebral hemorrhage(P ＞ 0.05).Conclusions Ginkgolide B may lessen neuronal cell apoptosis by means of inhibition of free radical production and inflammatory reactions after intracerebral hemorrhage.