针刺督脉经穴和夹脊穴对脊髓损伤大鼠神经干细胞的作用

Effect of acupuncture on Governing Vessel and Jiaji （Ex-B2） acupoints on endogenous neural stem cells following spinal cord injury in rats

目的探讨针刺督脉经穴和夹脊穴对脊髓损伤大鼠神经功能恢复和内源性神经干细胞增殖的影响。方法48只6周龄SD大鼠以改良Allen WD法制作成脊髓损伤模型，并按随机数字表法分为A组和B组，每组又分为14d、30d、60d和90d4个时间点。A组每天针刺督脉经穴和夹脊穴1次．B组不针刺。各组在相应时间点取材前连续3d腹腔注射BrdU标记增殖的内源性神经干细胞。各组分别在术后1d、14d、30d、60d和90d用Tarlov评分和IP评分评价双后肢功能，术后14d、30d、60d和90d评分后取材进行抗BrdU免疫组化染色，观察各切片BrdU阳性细胞数目及分布情况。结果双后肢功能评分显示术后14d、30d、60d和90dA组双下肢功能改善与B组比较差异有统计学意义（P〈0．05），功能改善与治疗时间延长平行。免疫组化染色显示术后14d、30d、60d和90dA组BrdU阳性细胞数和B组比较差异有统计学意义（P〈0．05）。结论针刺督脉经穴和夹脊穴能促进脊髓损伤大鼠双下肢功能恢复和原位诱导脊髓内源性神经干细胞增殖。

Objective To observe the neurological recovery and endogenous neural stem cell proliferation in rats with spinal cord injury after acupuncture on Governing Vessel and Jiaji （Ex-B2） acupoints. Methods Forty-eight adult SD rats with severe spinal cord injury at the T10 level induced with the method of Allen were randomly divided into two groups （A and B）, and each was further randomized into 4 subgroups. In group A, acupuncture was performed once daily on the Governing Vessel and Jiaji （Ex-B2） points, which was not performed in group B. Bromodeoxyuridine （BrdU） was administered intraperitoneally twice daily 3 days prior to heart perfusion for labeling the proliferating endogenous neural stem cells. Open field and inclined plane tests were performed to test the fimction of the rear limbs of the rats at 1, 14, 30, 60, 90 days after the surgery. The 4 subgroups of rats were sacrificed on days 14, 30, 60 and 90 after the surgery, and the number of BrdU-positive cells and their distribution in the spinal cord was observed in each subgroup using immunohistochemistry with anti-BrdU. Results The rats in group A showed significantly improved neurological fimction than those in group B （P〈0.05）. The number of BrdU-positive ceils in group A was significantly different from that in group B at each of the time points （P〈0.05）. Conclusion Acupuncture on the Governing Vessel and Jiaji （Ex-B2） acupoints can promote the functional recovery of the rear limbs and induce the proliferation of endogenous neural stem cells in situ in rats following spinal cord injury.