督脉电针与神经干细胞移植联合应用促进大鼠受损伤脊髓组织产生神经生长活性物质

COMBINATION OF GOREMOR VESSEL ELECTROACUPUNCTURE AND TRANSPLANTED NEURAL STEM CELLS PROMOTES INJURED SPINAL CORD TISSUE PRODUCTING NERVE GROWTH ACTIVE SUBSTANCES

目的探讨督脉电针与神经干细胞移植联合应用能否促进受损伤脊髓组织产生神经生长活性物质。方法成年大鼠分为正常组、对照组、神经干细胞移植组(NSCs组)、督脉电针组(电针组)和督脉电针+神经干细胞移植组(电针NSCs组)。除正常组外均实施T10段脊髓全横断手术,其中电针组和电针NSCs组于术后5d进行电针治疗。分别于术后14 d和28 d取材,进行受损伤脊髓组织环一磷酸腺苷(cAMP)含量的放射免疫检测;用Westenblotting检测神经营养素-3(NT-3)、神经营养因子受体(Trk)以及生长相关蛋白-43(GAP-43)的水平。结果1.在术后14 d,NSCs组、电针组和电针NSCs组的脊髓损伤区cAMP含量较正常组有增加。但除电针组外,脊髓损伤区的上段或下段组织cAMP含量低于正常组。在术后28 d,电针NSCs组脊髓损伤区上段、下段组织cAMP含量仍保持较高水平。NSCs组和电针NSCs组的脊髓损伤区cAMP含量较正常组有所增加。2.NT-3、Trk及GAP-43的表达以电针组和电针NSCs组较为明显,对照组和NSCs组的3种蛋白表达均低于电针组和电针NSCs组。结论督脉电针能促进受损伤的脊髓组织产生cAMP;督脉电针与NSCs移植联合应用能保持受损伤的脊髓组织较高水平的cAMP,而且NT-3、Trk及GAP-43的含量也有增高的趋势。

Objective To explore whether combination of Goremor vessel electroacupuncture and transplanted neural stem cells could promote the completely transected spinal cord tissue producting some nerve growth active substances. Methods Fourty-five adult rats were divided into normal group, control group, neural stem cells transplanted group （NSCs group）, Goremor vessel electroacupunctural group （EC group） and Goremor vessel electroacupuncture plus NSCs transplanted group （EC ＋ NSCs group）. All animals （except for the normal group） were performed complete transection on T10 segment of spinal cord. The electroacupunctural treatment started at the fifth day after operation among EC group and EC ＋ NSCs group. The sacrifice time of rats was the 14d and 28d after operation respectively. The cyclic adenosine-monophosphate （cAMP） radioimmunoassay and Western blotting test for levels of Neuroltrophin-3 （NT-3）, neurotrophic factor recepto~ Trk and growth associated protein-43 （GAP-43） were made in the tissue of injured spinal cord. Results 1. Fourteen days after operation, cAMP concentration at cranial segment and caudal segment of injured spinal cord segment was lower in all groups than that of normal group, except for EC group, cAMP concentration at injured segment was increased slightly in EC group and EC ＋ NSCs group than that of normal group, Twenty-eight days after operation, cAMP concentration at cranial segment and caudal segment of injured spinal cord segment maintained a higher level in EC ＋ NSCs group, cAMP concentration at injured segment was increased slightly in NSCs group and EC ＋ NSCs group than normal group. 2. The expression of NT-3, Trk and GAP-43 was more prominent in EC group and EC ＋ NSCs group. Meanwhile, the expression of three proteins was lower in control group and NSCs group than that of EC group and EC ＋ NSCs group. Conclusion Goremor vessel electroacupuncture may promote the injured spinal cord tissue producting cAMP. The combination of Goremor vessel electroacupuncture and transplanted neural stem cells may maintain higher cAMP concentration and tend to increase the concentration of NT-3, Trk and GAP-43 in injured spinal cord tissue.