六味地黄膏膏摩对脑瘫幼鼠大脑皮层损伤修复及BDNF/TrkB表达的影响

Effects of Liuwei Dihuang Ointment Rubbing(六味地黄膏膏摩)on the Repair of Cerebral Cortex Injury and BDNF/TrkB Expression in Cerebral Palsy Rats

目的:观察六味地黄膏膏摩对脑瘫(CP)幼鼠大脑皮层损伤修复的影响,探讨其促进脑损伤修复的作用机制。方法:采用左侧颈总动脉凝断合并缺氧的方式制备CP模型,并将其随机分为模型对照组、推拿组、涂膏组及膏摩组,同时设立假手术对照组。术后第1天开始,推拿组给予摩肾俞的推拿干预;涂膏组给予六味地黄膏外涂干预;膏摩组给予六味地黄膏外涂及摩肾俞相结合的膏摩干预;假手术对照组及模型对照组在其余3组接受干预的同时给予禁食处理。以上干预或禁食措施1次/d,连续28 d。采用尼氏染色观察幼鼠皮层神经元的形态及数量,并测定外周血清及大脑皮层中脑源性神经营养因子(BDNF)/酪氨酸激酶受体B(TrkB)的蛋白表达水平。结果:推拿组、涂膏组及膏摩组幼鼠大脑皮层正常形态神经元数量均多于模型对照组(P<0.01),膏摩组多于推拿组(P<0.05)及涂膏组(P<0.01),3组幼鼠大脑皮层神经元的形态分布较模型对照组均有所改善。膏摩组幼鼠血清BDNF含量高于模型对照组(P<0.05),大脑皮层BDNF表达水平高于模型对照组、推拿组、涂膏组(P<0.01);但其大脑皮层TrkB表达水平低于模型对照组(P<0.05)。推拿组、涂膏组幼鼠血清及大脑皮层BDNF/TrkB表达水平与模型对照组比较,差异均无统计学意义(P>0.05)。结论:推拿、涂膏或膏摩均能对脑瘫幼鼠大脑皮层的损伤起到修复作用,其中膏摩效果最佳;六味地黄膏膏摩的促大脑皮层损伤修复机制可能与BDNF/TrkB或其他神经营养因子有关。

Objective:To observe the effect of Dihuang Ointment Rubbing on the repair of cerebral cortex in young rats with cerebral palsy(CP),and to explore its mechanism of promoting the repair of brain injury.Methods:The CP model was prepared by coagulation of the left common carotid artery and hypoxia,and it was randomly divided into model control group,Tuina group,ointment-smearing group and ointment-rubbing group,and a sham operation control group was set up.From the first day after operation,the Tuina group was given Shenshu-rubbing,the ointment-smearing group was given the intervention of Liuwei Dihuang Ointment,the ointment-rubbing group was treated with Liuwei Dihuang Ointment and Shenshu-rubbing.The sham operation control group and the model control group were given fasting treatment while receiving intervention in the other three groups.The above intervention or fasting measures once a day for 28 consecutive days.Nissl staining was used to observe the morphology and number of cortical neurons in young rats,and to determine the protein expression levels of brain-derived neurotrophic factor(BDNF)/tyrosine kinase receptor B(TrkB)in peripheral serum and cerebral cortex.Results:The number of normal morphological neurons in the cerebral cortex of young rats in Tuina group,ointment-smearing group and ointment-rubbing group were more than that of the model control group(P<0.01),and the ointment-rubbing group was more than the Tuina group(P<0.05)and the ointment-smearing group(P<0.01).The morphological distribution of neurons in the cerebral cortex of young rats in the three groups was improved compared with the model control group.The serum BDNF content of young rats in ointment-rubbing group was higher than that of the model control group(P<0.05),and the expression level of BDNF in the cerebral cortex was higher than that of the model control group,Tuina group,and ointment-smearing group(P<0.01),however,the expression level of TrkB in the cerebral cortex was lower than that in the model control group(P<0.05).There was no significant difference between the expression levels of BDNF/TrkB in serum and cerebral cortex of young mice in Tuina group,ointment-smearing group and model control group(P>0.05).Conclusion:Tuina,ointment-smearing and ointment-rubbing can all play a role in repairing cerebral cortex damage in young rats with cerebral palsy,and the ointment-rubbing has the best effect.The mechanism of Liuwei Dihuang Ointment rubbing in promoting cerebral cortex damage and repair may be related to BDNF/TrkB or other neurotrophic factors.