电针对局灶性脑梗死大鼠皮质Slit2及HSPGs表达的影响

Effect of Electro-acupuncture on the Expression of Slit2 and HSPGs in Rats with Focal Cerebral Infarction

中文关键词:  脑缺血再灌注 电针 Slit2-Robo 硫酸肝素蛋白多糖

英文关键词:Cerebral ischemia-reperfusion Electroacupunture Slit2-Robo HSPGs

中文摘要:

      目的 观察局灶性脑梗死大鼠皮质轴突导向因子Slit2、硫酸肝素蛋白多糖(HSPGs)的表达和电针对其表达的影响，探讨电针对脑梗死后神经可塑性的作用机制。方法 将40只雄性SD大鼠随机分为正常组、模型组、非经穴电针组和电针组，每组10只。用线栓法栓塞模型组、非经穴电针组、电针组大鼠大脑中动脉1.5 h后恢复血流。电针组大鼠针刺“足三里”“曲池”， 非经穴电针组则针刺足阳明胃经、手阳明大肠经以外的非14经穴的左侧的其它穴位，1次/d，共14 d，模型组、正常组大鼠在自制容器中固定30 min。在14 d时用神经功能评分（mNSS）观察大鼠神经功能缺损；用尼氏染色观察大脑梗死灶周围组织形态学变化；用免疫荧光、免疫印迹检测大脑梗死灶组织Slit2和HSPGs的表达。结果 mNSS评分显示，正常组评分为0，低于电针组，电针组低于非经穴电针组，模型组最高，两两比较差异有统计学意义（ P<0.01）。尼氏染色显示，电针组可见尼氏体数量增多、排列整齐；非经穴电针组可见尼氏体数量增多，但排列紊乱；模型组尼氏体数量变少、排列紊乱且大脑水肿。免疫荧光和免疫印迹显示，电针组Slit2、HSPGs荧光强度和蛋白表达高于非经穴电针组（P<0.05），非经穴电针组荧光强度和蛋白表达高于模型组（ P<0.05），模型组荧光强度和蛋白表达低于正常组（P<0.05）。尼氏染色、免疫荧光、免疫印迹分析结果一致。结论 电针可以促进局灶性脑梗死大鼠皮质HSPGs及Slit2表达，从而促进脑梗死后神经功能的恢复，这可能是电针促进脑梗死后神经功能恢复的机制之一。

英文摘要:

      Objective To determine the expression of Slit2 and heparan sulfate proteoglycans (HSPGs) in cortex of rats with focal cerebral infarction and the effect of acupuncture (EA) on the expression of Slit2 and HSPGs. Methods 40 male Sprague Dawley (SD) rats were equally randomized into four groups：Control group，model group，non-acupoint EA group，and EA group. Thread-tying method was used in the model group, non-acupoint EA group and EA group to clog arteries and then open up after 1.5 h. Morphology changes of tissues around the infarction area were observed 14 days later by Nissl staining. The expressions of Slit2 and HSPGs in the ischemic brain tissues were detected by indirect immunofluorescence staining (IF) and Western blot (WB). Results Modified neurologicalseverity scores (mNSS) showed zero in the control group, lower than the score of EA group. The EA group had lower mNSS score than the non-acupoint group. The highest mNSS score appeared in the model group. Paired comparisons showed statistical differences （ P<0.01）. Nissl’s staining showed that EA group had increased Nissl bodies in alignment; non-acupoint EA group had increased and disordered Nissl bodies; model group had decreased and disordered Nissl bodies with edema in the brain. IF and WB showed that non-acupoint EA group had higher levels of Slit2 and HSPGs than model group ( P<0.05); EA group had higher levels of Slit2 and HSPGs than non-acupoint EA group ( P<0.05); model group had higher levels of Slit2 and HSPGs than control group ( P<0.05). The results of Nissl’s staining, IF and WB were consistent. Conclusion EA can enhance the expressions of Slit2 and HSPGs. This may be one of the mechanisms of EA promoting recovery of neural functions after cerebral infarction.