艾灸对脑缺血后内源性干细胞的增殖影响的实验研究

摘要

Objective To observe the effect of moxibustion on the proliferation of neural stem cells after cerebral ischemia- reperfusion in rat models.Method The middle cerebral artery occlusion (MCAO) model was developed by using modified Zea-Longa method. Seventy-two Wistar rats were randomized into group A (sham-operation group), group B (model group) and group C (moxibustion group). Each group was then further divided into 4 sub-groups according to different time points: 3 d, 7 d, 14 d, and 21 d, 6 rats in each sub-group. For each group, Baihui (GV 20) and bilateral Taiyang (EX-HN 5) were selected to study. The proliferated cells were marked by intraperitoneal injection of BrdU, and TTC staining and HE staining were adopted to detect the pathological changes of brain tissues after cerebral ischemia-reperfusion. Immunohistochemical method was used to dynamically detect the positive expressions of BrdU and Nestin cells in DG and SVZ zones at different time points, and microscope image analysis was taken to study the positive expressions.Result The neurological deficit score in group C was significantly lower than that in group B at each time point (P<0.05). The neurological deficit score in group C was significantly different from that in group B at the same time point (P<0.05). Moxibustion can up-regulate the expression of BrdU in SVZ and DG, and the increase started from the 3rd day and reached the peak on the 7th day. The increase of BrdU expression in group C was more significant than that in group B (P<0.05). Moxibustion can up-regulate the expression of Nestin in SVZ and DG, and the increase started from the 3rd day and reached the peak on the 7th day. The increase of Nestin expression in group C was more significant than that in group B (P<0.05).Conclusion Moxibustion can remarkably promote the recovery of nerve function and the proliferation of BrdU and Nestin after cerebral ischemia-reperfusion in rats.%目的：观察艾灸对大鼠脑缺血再灌注后内源性神经干细胞增殖的影响。方法选用Wistar大鼠72只,通过改良线栓法制备大脑中动脉局灶脑缺血再灌注模型(MCAO模型),随机分为A组(假手术组)、B组(模型组)和C组(艾灸组)。每组再分3 d、7 d、14 d、21 d 4个时间点,每个时间点6只。各组均取大鼠百会、太阳(双)穴进行实验研究。通过腹腔注射Brdu来标记增殖的细胞,根据TTC染色和HE染色来了解脑缺血再灌注后脑组织的病理学变化。通过免疫组织化学方法动态检测DG和SVZ区这两个神经生发中心增殖细胞Brdu和Nestin阳性细胞随时间点不同的表达变化,并采用显微图像分析系统对阳性结果进行分析。结果 C组各时间点神经功能缺损评分与B组比较,评分显著降低(P＜0.05)。C组同一时间点神经功能缺损评分与B组比较,差异均具有统计学意义(P＜0.05)。艾灸可以促进室管膜VZ区与海马DG区BrdU免疫阳性细胞表达增强,3 d时开始增加,7 d时达到高峰,与B组相比,C组阳性细胞数量显著增加(P＜0.05)。艾灸可以促进室管膜VZ区与海马DG区Nestin免疫阳性细胞表达增强,3 d时开始增加,7 d时达到高峰,与B组相比,C组Nestin阳性细胞数量显著增加(P＜0.05)。结论艾灸能显著促进脑缺血再灌注后大鼠神经功能的恢复以及Brdu、Nestin阳性细胞的增殖。