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目的研究益气活血补肾方“髓复康”对大鼠脑缺血损伤区和体外培养神经胶质瘢痕中硫酸软骨素蛋白多糖(CSPGs)表达的抑制作用。方法通过Koizumi法制作SD大鼠单侧大脑中动脉阻塞模型(MACO),将SD大鼠随机分为正常组、模型组、阳性对照组(激素组)、髓复康大剂量组、中剂量组和小剂量组,均在8,15和30 d 3个时间点取大鼠脑组织制作病理切片,通过免疫组化染色方法检测各组脑缺血损伤区硫酸软骨素蛋白多糖表达量的差异;建立体外培养神经胶质瘢痕模型,以血清药理学的方法选择不同浓度的含药血清,使用免疫组化染色方法分别在培养12,24,48 h后定量检测神经胶质细胞硫酸软骨素蛋白多糖的表达量。结果整体动物实验和体外培养的神经胶质细胞中,中药大剂量及中剂量组神经胶质细胞的反应性增生较轻,硫酸软骨素蛋白多糖表达最弱,均与模型组形成极显著差异(P<0.01),小剂量组的硫酸软骨素蛋白多糖表达也低于模型组,与之形成显著差异(P<0.05)。结论 “髓复康”可以抑制脑缺血损伤区硫酸软骨素蛋白多糖的表达,改善脑缺血损伤区轴突再生微环境,可能是其促进脑缺血损伤后轴突再生的机制之一。
Objective To investigate the inhibitory effect of Yiqi Huoxue Bushen Fang “Zu Fu Kang ” on the expression of CSPGs in cerebral ischemic injury area and glial scar in vitro. Methods The middle cerebral artery occlusion model (MACO) of SD rats was made by Koizumi method. The SD rats were randomly divided into normal group, model group, positive control group (hormone group) And low-dose group, all rats were sacrificed at 8, 15, and 30 days to make pathological sections. The expression of chondroitin sulfate proteoglycan in each group was detected by immunohistochemical staining. The glial scar model was established in vitro. Serum pharmacology method was used to select serum containing different concentrations. Immunohistochemical staining was used to detect the expression of glial cell chondroitin sulfate proteoglycan The amount of expression. Results In the whole animal experiment and in vitro cultured glial cells, the glial cells in the high-dose and medium-dose groups had less reactive hyperplasia and weakest expression of chondroitin sulfate proteoglycan, all of which were significantly different from the model group ( P <0.01). The expression of chondroitin sulfate proteoglycan in the low dose group was also lower than that in the model group (P <0.05). Conclusion “Nifukang” can inhibit the expression of chondroitin sulfate proteoglycan in cerebral ischemic injury area and improve the microenvironment of axonal regeneration in ischemic area, which may be the mechanism of axon regeneration after cerebral ischemic injury one.