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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0.3mL/(kg·d)的总量分4次(每6h1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8,24,72h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。
BACKGROUND: A large number of cells die in apoptosis after secondary injury after spinal cord injury. OBJECTIVE: To investigate the effect of local injection of Salvia miltiorrhiza injection on spinal cord necrosis and apoptosis after acute spinal cord injury. Design: Randomized controlled trials. Unit: Clinical Medicine Experimental Center, People’s Hospital Affiliated to Yunyang Medical College. Subject: 44 Chinese rabbits of 4 to 5 months of age, male or female, body weight 2.0 to 2.5 kg. MATERIALS: Selected trials were performed at the Clinical Medical Experimental Center of the Affiliated People’s Hospital of Yunyang Medical College from June 2002 to July 2003. Randomly divided into 2 groups, Salvia miltiorrhiza group and control group, 22 in each group. Both groups of animals were treated with modified Allen’s method to induce incomplete spinal cord injury in rabbits. Salvia miltiorrhiza group was divided into four injections (in every six hours) at a total volume of 0.3 mL/(kg·d) from the subdural catheter, and the control group was given an equal volume of normal saline. Animals were sacrificed at 8, 24, and 72h after injury. Pathological and histomorphological observations, detection of superoxide dismutase and malondialdehyde, and apoptosis-inhibiting gene B-cell lymphoma-2 (Bcl-2) positive cells were performed. Testing. MAIN OUTCOME MEASURES: Apoptosis index and apoptosis rate of spinal cord injury area. RESULTS: Forty-four rabbits were involved in the analysis of the results. 1 Apoptosis results: The apoptotic index of Danshen group was significantly less than that of the control group (13.10±1.38, 20.39±2.96, t=4.101, P<0.01); the apoptosis rate was significantly lower than that of the control group [(9.67±1.09) %,(14.68±2.81)%,t=4.072,P<0.01]; The expression of Bcl-2 was higher than that of the control group [(19.12±4.74)/mm2,(13.37±3.68)/mm2,t=2.347, P<0.01]. 2 The content of superoxide dismutase: Salvia miltiorrhiza group was higher than the control group [(136.20±13.64) NU/mL, (101.70±15.24) NU/mL, t=4.132, P<0.01]. 3 Malondialdehyde content: Salvia miltiorrhiza 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]. 4 Degeneration and necrosis of neurons and nerve fibers: Salvia miltiorrhiza group was lighter than the control group. Conclusion: Local administration of Salvia Miltiorrhiza can reduce the local apoptosis of spinal cord injury, and inhibit and reduce the cell necrosis after acute spinal cord injury.