| ObjectiveBased on the theory of "kidney-brain related",Liuwei Dihuang ointment rubbing was used in cerebral palsy rats.The effects of Liuwei Dihuang ointment rubbing on the cortical injury repair and BDNF/TrkB expression were observed.This study was aim at providing the evidence of Liuwei Dihuang ointment rubbing for CP.Methods1.Literature ReviewBy using literature collection and data mining methods,the origin and development of the theory of "kidney-brain related" was discussed.The substance of TCM on the etiology and pathogenesis of CP was explored.The syndrome differentiation and acupoint selection rules of Qilu pediatric Tuina therapy for CP were sorted out.The theoretical basis and advantages of Liuwei Dihuang ointment rubbing therapy for CP were analyzed.2.Experimental Study(1)Experimental animals: After modeling and other treatment measures,50 young rats were randomly divided into model control group,sham operation control group,Tuina group,pasting application group and Liuwei Dihuang ointment rubbing group,with10 rats in each group.(2)Intervention methods: Finger rubbing on Shenshu was used in the Tuina group.In the pasting group,Liuwei Dihuang ointment was applied to Shenshu region.In the Liuwei Dihuang ointment rubbing group,the Liuwei Dihuang ointment was applied to Shenshu point and finger rubbing was performed then.The interventions of the three groups were performed once a day for 28 days.The rats in model control group and sham operation control group were given fasting treatment only.(3)Index detection:Serum samples,cerebral paraffin sections and cortical proteins were extracted and prepared after the rats were anesthetized and sacrificed.The morphology and number of neurons in cerebral cortex were observed by Nissl staining.Serum brain-derived neurotrophic factor(BDNF)and tyrosine kinase receptor B(TrkB)were detected by ELISA test.Immunofluorescence staining was used to observe the fluorescence level of BDNF/TrkB in cerebral cortex.Western blot was used to detect BDNF/TrkB protein expression levels in cerebral cortex.Results1.Literature Review(1)The theory of "kidney-brain related" originated in the Warring States,Qin and Han Dynasties and first appeared in the Huangdi Neijing,which has been constantly enriched and improved with the development of Traditional Chinese medicine and applied in the syndrome differentiation.(2)Congenital endowment deficiency,renal medullary insufficiency and cerebral medullary loss were the main etiology and pathogenesis of CP.(3)Qilu pediatric Tuina treated CP based on syndrome differentiation under the guidance of "kidney-brain related" theory.Shenshui,Erren Shangma and Shenshu points were adopted as the core of CP treatment,but there was no animal surface positioning of Shenshui and Erren Shangma.(4)The Liuwei Dihuang ointment rubbing method was composed of Shenshu rubbing and Liuwei Dihuang ointment.Liuwei Dihuang ointment rubbing could give full play to the dual effects of Tuina and medicine,and has the function of nourishing Yin,tonifying kidney and filling Jing and Sui.It solved the problem of drug taking difficulty in children with CP,and was suitable for children with CP.2.Experimental Study(1)Effects of Liuwei Dihuang ointment rubbing on the morphology and number of cerebral cortex neurons in CP rats.Compared with the model control group,the morphology and distribution of cortical neurons in the Liuwei Dihuang ointment rubbing group,Tuina group and pasting group were improved.The staining depth and transparency were more consistent.There were more Nissl bodies,and the number of neurons in normal shape was higher than that in the model control group,the difference was highly statistically significant(P < 0.001).The morphology and distribution of cortical neurons in Liuwei Dihuang ointment rubbing group,Tuina group,pasting group were all poor,and the staining depth and transparency were different.However,compared with the Tuina group and the pasting group,the number of normal cortical neurons in the Liuwei Dihuang ointment rubbing group increased,and the difference was statistically significant(P < 0.05,P < 0.01).There was no significant difference in the number of normal cortical neurons between Tuina group and pasting group(P >0.05).(2)Effect of Liuwei Dihuang ointment rubbing on serum and cortical BDNF/TrkB expression in CP young rats.Serum BDNF content and cortical BDNF protein expression levels in the Liuwei Dihuang ointment rubbing group were all higher than those in the model control group(P < 0.05,P < 0.001).The serum BDNF content and cortical protein BDNF expression levels in the Tuina group and the pasting group were compared with the model control group,and the difference was not statistically significant(P >0.05).There was no significant difference in serum BDNF content between Liuwei Dihuang ointment rubbing group,Tuina group,pasting group(P >0.05).Compared with Tuina group and pasting group,the expression level of BDNF protein in the cortex of the ointment rubbing group was increased,and the difference was highly statistically significant(P <0.001).There was no significant difference in cortical BDNF protein expression between Tuina group and pasting group(P >0.05).The serum TrkB contents of Liuwei Dihuang ointment rubbing group,Tuina group and pasting group were compared with the model control group,and the difference was not statistically significant(P >0.05).Compared with the model control group,the expression level of TrkB protein in the cortex of ointment rubbing group was decreased,and the difference was statistically significant(P <0.05).The expression levels of TrkB protein in Tuina group and pasting group were not statistically significant compared with the model control group(P >0.05).Compared with Tuina group,the serum TrkB content in the Liuwei Dihuang ointment rubbing group was decreased,and the difference was statistically significant(P<0.05).There was no statistically significant difference in serum TrkB content when ointment rubbing group and Tuina group compared with pasting group(P >0.05).Compared with the Tuina group and the pasting group,the expression level of TrkB protein in the cortex of ointment rubbing group was decreased,and the difference was statistically significant(P<0.05).There was no significant difference in TrkB protein expression between the Tuina group and the pasting group(P >0.05).Conclusions1.The Liuwei Dihuang ointment rubbing therapy was based on "kidney-brain related" theory,CP etiology and pathogenesis and Qilu pediatric Tuina therapy for CP.Liuwei Dihuang ointment rubbing has the effect of nourish kidney to benefit brain,which is feasible and advantageous for CP.2.Liuwei Dihuang ointment rubbing,singly Tuina and singly pasting could all play a certain role in repairing the cortical neuron injury of CP rats,and the effect of ointment rubbing was the best.2.Liuwei Dihuang ointment rubbing could enhance the expression of BDNF in the serum and cerebral cortex of CP rats,but had no significant effect on the expression of TrkB in the serum.While Tuina and Liuwei Dihuang ointment pasting had no significant effect on the expression of BDNF/TrkB in serum and cerebral cortex of CP rats.Liuwei Dihuang ointment rubbing inhibited the expression of TrkB in the cerebral cortex.The reduce of the expression level of TrkB in cortex may be related to the phosphorylation of TrkB. |
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