两种督脉电针对大鼠脊髓受损伤神经元存活的影响

Effect of two kinds of Governor Vessel electro-acupuncture on the survival of spinal neurons after rat spinal cord injury

目的探讨脊髓损伤区近端督脉2个穴位电针和脊髓损伤区远、近端督脉4个穴位电针对全横断脊髓损伤大鼠腰段脊髓神经元存活与凋亡的影响。方法雌性SD大鼠30只,随机分为对照组、近端督脉穴电针组(EA2组)和远近端督脉穴电针组(EA4组),每组10只大鼠,其中每组各5只分别应用于免疫荧光染色和Western blot检测。在所有大鼠脊髓第10胸段(T10)进行全横断损伤手术。EA2组在脊中(GV9)和至阳(GV6)督脉穴位入针,EA4组在GV9、GV6、腰腧(GV4)和长强(GV1)督脉穴位入针。在术后第3 d开始电针,隔天1次,每次20 min。术后14d后将3组动物取材进行后续实验。Western blot检测脊髓组织活化型半胱天冬酶-3的表达情况。结果与对照组和EA 2组相比,EA 4组可增加脊髓腰段神经元存活数量、减少脊髓腰段神经元凋亡率和下调脊髓腰段组织表达活化型半胱天冬酶-3(P<0.05)。结论脊髓损伤区远近端督脉4个穴位电针要比脊髓损伤区近端督脉2个穴位电针更有效地促进受损伤脊髓腰段神经元存活和减少其凋亡,下调受损伤脊髓腰段组织表达凋亡相关蛋白。

Objective To investigate the effect of two kinds of Governor Vessel electro-acupuncture （EA） on the survival of spinal neurons after rat spinal cord injury （SCI）. Methods Thirty female Sprague-Dawley （SD） rats were ran- domly divided into the control group, EA2 group and EA4 group. Each group was 10 rats （5 rats in each group for TUNEL and immunofluorescence staining or Western blot detection, separately）. SCI model was established by transected complete- ly spinal cord at the level T10. At 3rd day after operation, EA was applied to Zhiyang （GV9） , Jizhong （GV6） , Yaoshu （GV2） and Changqiang （GV1） four acupoints in the EA4 group of rats, while the rats of the EA2 group were performed by EA in GV9 and GV6 two acupoints. There, was no EA in the rats of the control group. At 14th day after operation, all rats in three groups were carried out for next experimental stages. Results The results showed that the number of neurons in spinal cord lumbar segment was evidently increased in the EA4 group, compared with the control group and EA2 group （P〈0.05）. The apoptosis percentage of neurons in spinal cord lumbar segment was evidently decreased, and the activated caspase-3 ex- pressing in spinal cord lumbar segment tissue was obviously downregulated in the EA4 group, compared with the control group and EA2 group （P〈0.05）. Conclusion The effects of the Governor Vessel electro-acupuncture four acupoints on en- hancing the survival of neurons, decreasing the apoptosis of neurons and downregulating the tissue expressing activated cas- pase-3 in spinal cord lumbar segment are more obvious in rat spinal cord injury.