督脉电针对急性脊髓损伤6h后大鼠差异蛋白质组学的机理研究

Influence of GV electroacupuncture on different expressed proteins in rats with spinal cord injury in 6 hours

目的确定急性脊髓损伤(SCI)后差异表达蛋白,初步探讨其功能,从差异蛋白组学角度研究SCI的病理机制,以及电针对该病的治疗作用。方法大鼠随机分为空白组、假手术组、模型组、药物组、电针组,每组15只大鼠,共75只。采用自制的改良式Allen’s打击装置造模。术后6h取脊髓损伤区2cm的脊髓组织称重,保存。对样本蛋白进行提取、定量、蛋白双向电泳图像分析,找出差异点进行质谱分析和数据库检索。结果成功鉴定出6种蛋白质:假定蛋白LEF3-IAP2、泛素羧基末端水解酶、先驱B环蛋白3、假定蛋白ORF F-93、角蛋白16、泡溶酶蛋白。结论通过本动物实验可见,针刺对早期脊髓损伤具治疗效果。鉴定出的蛋白质可能通过引起炎症细胞浸润、神经细胞凋亡等途径参与了SCI的损伤及修复过程。

Objective To find different expressed proteins of acute spinal cord injury（SCI） and discuss the function of them to study the mechanism of SCI. To observe the effect of GV electroacupuncture on SCI. Methods Seventy-five rats were divided into five groups randomly： Blank Group, Sham Operated Group, Model Group, Drug Group and EA Group, with 15 rats in each group. The model rats were made a spinal cord contusion by using a modified Allen＇ s method.. After SCI EA Group was treated by electroacupuncture at ＂mingmen＂ （DU4） and ＂dazhui＂ （DU14） points; Drug Group was treated by methylprednisolone（MP）. Six hours after SCI, the rats were put to death and 2 cm tissue of spinal cord around the injured site were got out and weighed. The proteins were extracted and quantitated. The different expressed proteins were searched by using two-dimensional gel electrophoresis, analyzed by mass spectrum and searched in the data-base. Results Six different expressed proteins were identified： 1. Hypothetical 34.4 kd protein in lef3-iap2 intergenic region; 2. Ubiquitin carboxyl-terminal hydrolase isozyme L1 ;3. Balbiani ring protein 3 precursor; 4. Hypothetical 11.0 kDa protein; 5. Keratin, type Ⅰ cytoskeletal 16; 6. Vesicle-fusing ATPase. Conclusion Acupuncture has therapeutic effect on SCI. The above different expressed proteins take part in scathe and repair through arosing inflammatory cells infiltration and nerve cells apoptosis.