电针联合强制性运动对脑缺血大鼠GAP-43和GFAP表达的影响

Effects of electroacupuncture and constraint-induced movement therapy on the expression of GAP-43 and GFAP after middle cerebral artery occlusion in rats

目的:通过观察电针联合强制性运动对局灶性脑缺血大鼠灶周生长相关蛋白-43(GAP-43)及胶质纤维酸性蛋白(GFAP)表达的影响,探讨脑缺血后电针联合强制性运动疗法促进神经功能恢复的作用及机制。方法:将SD雄性大鼠采用线栓法制作大脑中动脉阻塞(MCAO)局灶性脑缺血模型,随机平均分为模型组、强制性运动(CIMT)组、电针组和电针联合CIMT组,每组以7、14、21、28和35d为时间点再随机平均分为5个亚组,同时设立假手术组。分别行相应治疗,以Morris水迷宫试验进行神经功能评价,分别于相应时间点取脑采用免疫组织化学方法观察每个时间点脑梗死灶周围GAP-43、GFAP阳性细胞数,Westernblot法检测脑组织GAP-43、GFAP蛋白表达。结果:与假手术组比较,模型组潜伏期和穿台次数显著高于假手术组,差异有统计学意义(P<0.001)。与模型组比较,CIMT组和电针组潜伏期和穿台次数比较,差异无统计学意义(P>0.05);电针联合CIMT组潜伏期和穿台次数显著低于模型组,也显著低于CIMT组和电针组,差异均有统计学意义(P<0.001);电针组与CIMT组比较,差异无统计学意义(P>0.05)。与模型组进行比较,在14、21、28d时电针联合CIMT组GAP-43表达显著上调,而GFAP表达显著下降,差异有统计学意义(P<0.05);且电针联合CIMT组GAP-43阳性细胞数多于CIMT组和电针组,差异有统计学意义(P<0.05);电针联合CIMT组GFAP阳性细胞数少于CIMT组和电针组,差异有统计学意义(P<0.05);在35d缺血区周围GAP-43表达增加、GFAP表达减少,但与CIMT组和电针组比较差异无统计学意义(P>0.05)。结论:电针联合CIMT可促进脑梗死灶周围GAP-43的表达,抑制GFAP的表达,从而改善大鼠神经功能。

Objective： To observe the change of electroacupuncture and CIMT on the neural function and the expression of GAP-43 and GFAP after middle cerebral artery occlusion in rats and investigate the effect of electroacupuncture and CIMT on the neural function. Methods： MCAO was established by nylon monofilament. These rats were randomly divided into 4 group： model group, CIMT group, electroacupuncture group, and electroacupuncture with CIMT group. Then each group was randomly divided into 5 subgroups by 7 d, 14 d, 21 d, 28 d, and 35 d. Meanwhile to design a sham-operation group. The corresponding treatment was performed respectively. The evaluations included the behavior scores and Morris water maze experiment, both was to compare the recovery of neural physiological function at different time points abovementioned. Immuno-histochemical technique was used to observe the positive cells of GAP-43 and GFAP in cortical ischaemic lesions of the rats in each group at different time points. The protein expression of GAP-43 and GFAP were detected by Western blot at day after ischemia. Results： The expression of GAP-43 in gray matter of rats were obviously increased in electroacupuncture with CIMT group compared with he model group, CIMT group, and acupuncture group after 14 d, 21 d, 28 d（P〈0.05）, The expression of GFAP in gray matter of rats were obviously decreased in electroacupuncture with CIMT group compared with CIMT group, electroacupuncture group after 14 d, 21 d, 28 d（P〈0.05）, but no significant difference was found between CIMT group and acupuncture therapy group after 35 d（P〈0.05）. Conclusion： Electroacupuncture combined with CIMT can improve neurological function of focal cerebral ischemia rats, which may be associated with the increase of GAP-43 and descrease of GFAP in peri-infarct regions.