椎管内局部灌注丹参注射液对急性脊髓损伤的保护

背景脊髓损伤后的继发性损伤期大量细胞以凋亡的方式死亡.目的探讨中药丹参注射液局部灌注对急性脊髓损伤后脊髓细胞坏死和调亡的影响.设计随机对照实验.单位陨阳医学院附属人民医院临床医学试验中心.对象4～5个月龄的一级中国白兔44只,雌雄不限,体质量2.0～2.5kg.材料选择实验于2002-06/2003-07在郧阳医学院附属人民医院临床医学试验中心完成.随机分成2组,丹参组和对照组,每组22只.两组动物均以改良Allen法造成兔不完全性脊髓损伤的模型.丹参组术后按0.3 mL/(kg·d)的总量分4次(每6 h 1次)从硬膜下导管推入丹参注射液,对照组推入等量生理盐水.注射至损伤后8,24,72 h分别处死动物,进行病理、组织形态学观察,过氧化物歧化酶和丙二醛检测,凋亡抑制基因B细胞淋巴瘤-2(Bcl-2)阳性细胞数检测.主要观察指标脊髓损伤区细胞凋亡指数和细胞凋亡率.结果44只兔均进入结果分析.①细胞凋亡结果丹参组的凋亡指数明显少于对照组(13.10±1.38,20.39±2.96,t=4.101,P＜0.01);细胞凋亡率明显低于对照组(9.67±1.09)%,(14.68±2.81)%,t=4.072,P＜0.01];Bcl-2的表达高于对照组(19.12±4.74)个/mm2,(13.37±3.68)个/mm2,t=2.347,P＜0.01].②过氧化物歧化酶含量丹参组高于对照组(136.20±13.64)NU/mL,(101.70±15.24)NU/mL,t=4.132,P＜0.01].③丙二醛含量丹参组低于对照组(1.27±0.22)nmol/mL,(2.54±0.69)nmol/mL,t=4.309,P＜0.01].④神经元和神经纤维变性及坏死丹参组轻于对照组.结论丹参局部灌注后,减少了脊髓损伤局部的细胞凋亡,抑制和减轻了急性脊髓损伤后细胞坏死.

Protection of local intraspinal infusion With danshen injection against acute spinal cord injury

BACKGROUND: A large amount of cell dies of apoptosis in secondary injury stage after spinal cord injury.OBJECTIVE: To probe into the influence of local intraspinal infusion with danshen injection on necrosis and apoptosis of spinal cell after acute spinal cord injury.DESIGN: Randomized controlled experiment was designed.SETTING: Clinical Medical Trial Center of People' s Hospital Affiliated to Yunyang Medical College.MATERIALS: Forty-four Chinese white rabbits of first grade were employed, aged varied from 4 to 5 months, of either sex, mass weighted varied from 2.0 to 2.5 kg.INTERVENTIONS: The experiment was performed in Clinical Medical Trial Center of People' s Hospital Affiliated to Yunyang Medical College from June 2002 to July 2003. Two groups were randomized, named danshen group and the control group, 22 rabbits for each. In both groups, modified Allen method was used to prepare the model of incomplete spinal cord injury. In danshen group, danshen injection was infused from subdural catheter for 4times at 0. 3 mL/kg per day of the total amount (once every 6 hours). In the control group, physiological saline of same dose was injected. The animals were sacrificed at the 8th, 24th and 72nd hours successively after injection for pathological and histomorphologic observation, peroxide dismutase and malondialdehyde determination and determination of positive cell count of B-cell lymphoma-2 (Bcl-2), the apoptosis-inhibition gene.MAIN OUTCOME MEASURES: Cell apoptotic index and cell apoptotic rate in the region of spinal cord injury.RESULTS: Forty-four rabbits entered result analysis for all. ① Results of cell apoptosis: Apoptotic index in danshen group was less remarkably than that in the control group(13.10 ± 1. 38, 20.39±2.96, 4.101, P ＜0.01); cell apoptotic rate was lower remarkably than the control group (9.67 ± 1.09)%,(14.68±2.81)%, t=4.072, P ＜0.01] and Bcl-2 expression was more than that in the control group (19. 12 ± 4.74) /mm2, ( 13.37 ± 3.68) /mm2,t = 2. 347, P ＜ 0.01 ]. ② POD content: The result in danshen group was higher thanthe control group (136.20 ± 13.64) NU/mL, (101.70 ± 15.24) NU/mL,t = 4. 132, P ＜ 0.01 ]. ③ Malondialdehyde content: The result in danshen group was lower than the control group (1.27 ± 0. 22) nmol/mL,(2.54±0.69) nmol/mL, t=4.309, P ＜0.01] . ④Degeneration and necrosis of neuron and neural fiber: The result in danshen group was milder than the control group.CONCLUSION: After local infusion with danshen(Radix Salviae Miltiorrhizae), cell apoptosis was decreased in local spinal cord injury and cell necrosis was inhibited and alleviated after acute spinal cord injury.