首页|期刊导航|中国神经再生研究（英文版）|Inhibitory effect of electro-acupuncture on apoptosis after spinal cord injury
中国神经再生研究（英文版）2006，Vol.1Issue(3)：239-243,5.
Inhibitory effect of electro-acupuncture on apoptosis after spinal cord injury
Inhibitory effect of electro-acupuncture on apoptosis after spinal cord injury
摘要
Abstract
BACKGROUND: Acupuncture and electro-acupuncture (EA) as complementary and alternative medicine have been applied for neurological recovery after spinal cord injury (SCI), but the mechanism of treatment is unclear yet.OBJECTIVE: To examine and compare the effects of EA with methylprednisolone (MP) on apoptosis and expression of caspase-3 mRNA and protein in early SCI, in an attempt to provide experimental clues for clinical use of EA in the treatment of early SCI.DESIGN: A randomized control trial.SETTING: Department of Anatomy, Second Military Medical University of Chinese PLA.MATERIALS: Forty-eight adult male Sprague-Dawley (SD) rats aged 12 weeks and weighing (250±20) g were used. The experimental procedures were performed in accordance with the animal care guidelines of the National Institute of Health (NIH). Equipments and medicine: Methylprednisolone sodium succinate (Pharmacia & Upjohn N.V./S.A., Puurs, Belgium); sodium pentobarbital (Sigma Chemical Co., St. Louis, MO); G-6805-2 Multi-Purpose Health Device (Shanghai Medical Instruments High-TECH Co., Shanghai, China); POD kit (Roche Molecular Biochemicals, Mannheim, Germany); ISH Detection Kit (Wuhan Boster Biological Technology Ltd., Wuhan, China); ABC kit (Vector Laboratories, Burlingame, CA); light microscope (×200, Olympus,Tokyo, Japan); HPIAS-1000 color High-definition pathological image analysis system (Qianping Imaging Engineering Company of Wuhan Tongji Medical University).METHODS: The experiment was carried out in Department of Anatomy, Second Military Medical University of Chinese PLA between October 2004 and March 2006. ① The SCI model was established by modified Allen's method on T10. The force applied in traumatization was 50 g·cm. ② Forty-eight SD rats were equally randomized into four groups: sham operation (SO) group, in which animals received laminectomy only; model control (MC) group as a negative control, in which animals did not receive any treatment; EA treatment (EA)group, in which animals were treated with electro-acupuncture immediately after SCI, and MP treatment (MP)group, in which animals were treated with 30 mg/kg methylprednisolone sodium succinate through the tail vein immediately after SCI. Electro-acupuncture treatment was performed for EA group at 8 hours and 20 hours after SCI. The needles were inserted at a depth about 5-6 mm into the locus of the Dazhui and Mingmen. Asymmetric waves were used for electro-acupuncture at a stimulating frequency of 1 Hz. The total duration of EA stimulation was 30 minutes. ③ Six rats of each group were sacrificed at 6 hours and 24 hours after SCI. A 1.5 cm segment of the spinal cord encompassing the injury site was removed and prepared for experiment. ④ Cellular apoptosis was observed by terminal deoxynucleotidyl transferase (TDT)-mediated deoxyuridine triphosphate (dUTP)-biotin nick end labeling (TUNEL) method. Expression of caspase-3 mRNA and protein were estimated by in situ hybridization, immunohistochemistry and imaging analysis. ⑤ Measurement data among groups were compared with one-way analysis of variance and differences between groups were compared with q test.MAIN OUTCOME MEASURES: The effect of electro-acupuncture on TUNEL-positive cells, the expression of caspase-3 mRNA and protein.RESULTS: The forty-eight rats were involved in the final experiment. ① TUNEL-positive cells: TUNEL-positive cells in MP group and EA group were 29.00±8.27 and 30.17±6.85, respectively, at 6 hours after SCI; and were 1.00±0.89, 60.17±7.83, 6.67±3.56; 1.17±1.17, 69.50±8.50, 9.67±4.97, respectively at tip, intermediate zone and end of spinal cord at 24 hours after SCI; which were less than those in MC group (47.17±11.12,9.33±4.76, 92.00± 11.14, 22.50±5.43, P ＜ 0.01). Cells in MP group were close to those in EA group (P ＞ 0.05).② Expression of positive caspase-3 mRNA: At 6 hours after SCI, numbers of positive caspase-3 mRNA were 133.33±11.84, 117.17±11.70, 120.00±10.73, respectively in MC, MP and EA groups, which were more than those in SO groups (83.50±8.64, P＜ 0.01). At 24 hours after SCI, numbers of positive caspase-3 mRNA were 121.50±11.34, 96.16±8.18, 97.67±9.69, respectively in MC, MP and EA groups, which were more than those in SO groups (83.50±9.64, P ＜ 0.05-0.01). Numbers in MP group were close to those in EA group but less than those in MC group (P ＜ 0.05-0.01). ③ Expression of caspase-3 protein: Numbers in MC and MP groups were close to those in EA group (12.83±2.86, 10.00±3.58, 10.50±3.62) at 6 hours after SCI; numbers in MP and EA groups were 11.00±4.43 and 12.17±3.43 at 24 hours after SCI, which were less than those in MC group (17.83±4.92, P＜ 0.01, 0.05).CONCLUSION: EA may partially inhibit apoptosis and protect nerve after SCI by down-regulating expression of caspase-3 mRNA and protein.分类
医药卫生引用本文复制引用
Zhiying Zhang,Ruishan Dang,Chuansen Zhang,Liang Li,Congli Ren,Shuang Zhou,Eryu Chen..Inhibitory effect of electro-acupuncture on apoptosis after spinal cord injury[J].中国神经再生研究（英文版）,2006,1(3):239-243,5.基金项目
Supported by:a grant of the Science and Technology Commission of Shanghai Municipality, No. 03DZ19554-7 （）
the National Basic Research Program,No. 2005CB523306 （）
