电针秩边穴和水道穴对急性脊髓不完全损伤大鼠脑皮质代谢物的影响

Effect of electroacupuncture at Point BL54( Zhibian) and Point ST28( Shuidao) on cerebral cortex metabolites in rats with acute incomplete spinal cord injuries

目的：观察电针秩边穴和水道穴对急性脊髓不完全损伤大鼠脑皮质代谢物的影响。方法：将36只雄性 SD 大鼠随机分为假手术组、模型组、电针组，每组12只。采用改良 Allens 法将模型组和电针组大鼠制成 T10急性脊髓不完全损伤模型，假手术组大鼠手术摘除 T10椎板暴露硬脊膜后立即缝合。造模结束后，假手术组和模型组大鼠仅进行抓取和固定；电针组大鼠予以电针秩边穴和水道穴，每天1次，连续治疗1周。电针干预结束后断头处死所有大鼠，采集大鼠脑皮质和 T9～T10椎体对应节段脊髓1 cm。将取得的脊髓组织制成切片，HE 染色后光镜下观察脊髓组织形态；对取得的脑皮质进行磁共振波谱分析，比较3组大鼠脑皮质代谢物的浓度。结果：至处死前各组均无大鼠死亡。假手术组大鼠脊髓结构完整，神经细胞形态正常、分布均匀，细胞膜、细胞核及组织间隙均正常；模型组大鼠脊髓形态不完整，损伤区域有神经组织残缺，存在较多出血，组织疏松水肿，部分细胞核固缩，神经细胞数量明显减少，细胞周围基质消失呈空泡状；电针组大鼠脊髓形态结构基本完整，组织中散在血细胞，组织水肿较轻，神经细胞形态基本正常，空泡变性较轻。3组大鼠脑皮质中异亮氨酸、谷氨酸、N －乙酰天门冬氨酸、甘油磷脂胆碱、肌醇、维生素 C、乳酸、苏氨酸、肌苷浓度比较，组间差异均有统计学意义（1．63±0．20，1．13±0．12，1．35±0．01，F ＝43．195，P ＝0．000；9．51±0．40，8．66±0．74，9．94±1．25，F ＝6．728，P ＝0．004；2．54±0．22，2．21±0．19，2．38±0．45，F ＝3．332，P ＝0．048；9．47±1．26，8．10±1．34，9．38±0．88，F ＝5．070，P ＝0．012；3．66±1．10，3．17±0．89，4．55±1．81，F ＝3．365，P ＝0．047；2．45±0．33，2．15±0．21，2．34±0．17，F ＝4．388，P ＝0．020；9．03±1．56，10．01±0．63，9．22±1．01，F ＝3．562，P ＝0．040；4．12±0．28，3．21±0．13，3．69±0．44，F ＝4．972，P ＝0．013；6．35±0．96，9．18±0．81，8．38±0．34，F ＝45．032，P ＝0．000）。假手术组和电针组异亮氨酸、谷氨酸、N －乙酰天门冬氨酸、甘油磷脂胆碱、肌醇、维生素 C、苏氨酸浓度均高于模型组（P ＝0．000，P ＝0．000；P ＝0．003，P ＝0．007；P ＝0．001，P ＝0．035；P ＝0．017，P ＝0．011；P ＝0．045，P ＝0．023；P ＝0．016，P ＝0．026；P ＝0．000，P ＝0．001），乳酸和肌苷的浓度均低于模型组（P ＝0．017，P ＝0．030；P ＝0．000，P ＝0．000）；假手术组异亮氨酸和苏氨酸的浓度均高于电针组（P ＝0．009；P ＝0．010），肌苷浓度低于电针组（P ＝0．005）；假手术组和电针组谷氨酸、N －乙酰天门冬氨酸、甘油磷脂胆碱、肌醇、维生素 C、乳酸的浓度比较，组间差异均无统计学意义（P ＝0．322；P ＝0．469；P ＝0．850；P ＝0．106；P ＝0．325；P ＝0．684）。结论：电针急性脊髓不完全损伤大鼠的秩边穴和水道穴，有利于其脊髓损伤恢复，并可提高其脑皮质中异亮氨酸、谷氨酸、N －乙酰天门冬氨酸、甘油磷脂胆碱、肌醇、维生素 C、苏氨酸的浓度，降低乳酸和肌苷的浓度。

Objective：To observe the effect of electroacupuncture at Point BL54（Zhibian）and Point ST28（Shuidao）on cerebral cortex metabolites in rats with acute incomplete spinal cord injuries.Methods：Thirty -six male SD rats were randomly divided into sham-operated group,model group and electroacupuncture group,1 2 cases in each group.The T1 0 acute incomplete spinal cord injuries models were built by modified Allen’s method in rats of model group and electroacupuncture group,while T1 0 vertebral plates were removed to expose the endorachis and the incision were sewed in rats of sham -operated group.After the end of the modeling,the rats in sham -operated group and model group were merely catched and fixed,while the rats in electroacupuncture group were treated with electroacupuncture at Point BL54（Zhibian）and Point ST28（Shuidao）,once a day for consecutive 1 week.After the end of electroacupuncture intervention,all rats were executed with decollation and the cerebral cortexs and 1 cm spinal cord at T9 -1 0 vertebral segment were collected.The spinal cord tissues were sectioned for HE staining and the spinal cord tissue morphology were observed under light microscope.The cerebral cortexs were analyzed by using nuclear magnetic resonance spectroscopy,and the concentrations of cerebral cortex metabolites were compared between the 3 groups.Results：No rats died in each group before execution.The structure of spinal cord was complete and the nerve cells were uniformly distributed in normal shape and their cellular membrane,cellular nucleus and tissue spaces were normal in rats of sham -operated group.The shape of spinal cord was incomplete in rats of model group and the injured region of spinal cord presented with（1 ）incomplete nervous tissues,（2）massive bleeding,（3）tissue edema,（4）karyopycnosis in some nerve cells,（5）significant decrease of nerve cells and （6）vacuolization in cell matrixs.The shape and structure of spinal cord were basically complete in rats of electroacupuncture group,and the injured region of spinal cord presented with（1 ）sporadic blood cells,（2）slight tissue edema,（3）basically normal shape of nerve cells and （4）slight vacuolization in cell matrixs.There was statistical difference in the concentrations of isoleucine,glutamic acid,N -acetyl -aspar-tate,glycerophosphorylcholine,inositol,vitamin C,lactic acid,threonine and inosine in the cerebral cortex beween the 3 groups（1 .63 ＋/-0.20,1 .1 3 ＋/-0.1 2,1 .35 ＋/-0.01 ,F =43.1 95,P =0.000;9.51 ＋/-0.40,8.66 ＋/-0.74,9.94 ＋/-1 .25,F =6.728,P =0.004;2.54 ＋/-0.22,2.21 ＋/-0.1 9,2.38 ＋/-0.45,F =3.332,P =0.048;9.47 ＋/-1 .26,8.1 0 ＋/-1 .34,9.38 ＋/-0.88,F =5.070,P =0.01 2;3.66 ＋/-1 .1 0,3.1 7 ＋/-0.89,4.55 ＋/-1 .81 ,F =3.365,P =0.047;2.45 ＋/-0.33,2.1 5 ＋/-0.21 ,2.34 ＋/-0.1 7,F =4.388, P =0.020;9.03 ＋/-1 .56,1 0.01 ＋/-0.63,9.22 ＋/-1 .01 ,F =3.562,P =0.040;4.1 2 ＋/-0.28,3.21 ＋/-0.1 3,3.69 ＋/-0.44,F =4.972,P =0.01 3;6.35 ＋/-0.96,9.1 8 ＋/-0.81 ,8.38 ±0.34,F =45.032,P =0.000）.The concentrations of isoleucine,glutamic acid, N -acetyl -aspartate,glycerophosphorylcholine,inositol,vitamin C and threonine in the cerebral cortex were higher in sham -operated group and electroacupuncture group compared to model group（P =0.000,P =0.000;P =0.003,P =0.007;P =0.001 ,P =0.035;P =0.01 7,P =0.01 1 ;P =0.045,P =0.023;P =0.01 6,P =0.026;P =0.000,P =0.001 ）,and the concentrations of lactic acid and inosine were lower in sham -operated group and electroacupuncture group compared to model group（P =0.01 7,P =0.030;P =0.000,P =0.000）.The concentrations of isoleucine and threonine were higher in sham -operated group compared to electroacupuncture group（P =0.009;P =0.01 0）,and the concentrations of inosine were lower in sham -operated group compared to electroacupuncture group（P =0.005）.There was no statistical difference in the concentrations of glutamic acid,N -acetyl -aspartate,glycerophosphorylcholine, inositol,vitamin C and lactic acid in the cerebral cortex between sham -operated group and electroacupuncture group （P =0.322;P =0.469;P =0.850;P =0.1 06;P =0.325;P =0.684）.Conclusion：Electroacupuncture at Point BL54（Zhibian）and Point ST28 （Shuidao）is helpful to the recovery of impaired spinal cord of rats with acute incomplete spinal cord injury,and it can increase the concentration of isoleucine,glutamic acid,N -acetyl -aspartate,glycerophosphorylcholine,inositol,vitamin C and threonine in the cerebral cortex of rats.