The Clinical Effect Of Baishaoluoshi Decoction On Post-stroke Spasticity And Its Effect On The Synaptic Plasticity And BDNF/TrKB-KCC2 Pathway

Part 1 Literature researchObjectiveTo explore the main principle of diagnosis and treatment regularity in post-stroke spasticity and to offer theoretical foundation for Baishaoluoshi Decoction.MethodsUsing documentary method to gather and sort out the records of post-stroke spasticity from ancient to now,as“ZHONG GUO ZHONG YI GU JI ZONG MU”for catalog reference.SPSS 20.0 was used for the frequency analysis,factor analysis,and clustering analysis on the database established by the Epi Data 3.1.ResultsA total of 854 ancient books of Chinese medicine were viewed,among which 78books were selected,and 107 nowadays literatures were selected at the same time.The 5 typical syndrome waixieweihuan,zhengxuxiezhong,zhengxushiyang, xushijiaza,shixiezuzhi were summarized.Nowadays,Chinese medicine doctor hold the perspective that xuexuluoyu,xuexujinmaishiyang,and qixuekuixu,luomaiyuzu are the principal syndromes of post-stroke spasticity,furthermore,they proposed that Papaya,Lycopodium clavatum,Millettia are the mainly symptomatic treatment drugs.The pathogenesis of xuexuluoyu in post-stroke spasticity is gradually be taken seriously,and is considered the key pathogenesis nowadys.Based on the above conclusions,professor Zhou Shen proposed Baishaoluoshi Decoction to treat post-stroke spasticity.ConclusionXuexuluoyu is the key pathogenesis in post-stroke spasticity,accordingly we proposed Baishaoluoshi Decoction to treat it.Part 2 Clinical researchObjectiveTo further evaluate the clinical efficacy and security of Baishaoluoshi Decoction.MethodsThe patients of post-stroke spasticity were randomly divided into zhongyao group(Baishaoluoshi Decoction)and control group.The myodynamia was evaluated by Robert Lovett's method,the muscle tension of the upper limb was evaluated by Modified Ashworth score(MAS),the muscle tension of the lower limb was evaluated by Compopsite Spasticity Scale(CSS),the neurological deficit symptoms were evaluated by National Institute of Health stroke scale(NIHSS),and the daily living ability was evaluated by Modified Barther Index(MBI).ResultsBaishaoluoshi Decoction can increase the muscle strength of hemiplegic limb after cerebral infarction(P<0.05),ameliorate the spasm of upper limb(P<0.05),improve the daily living ability(P<0.05),but have no significant effect to the spasm of lower limb(P>0.05).In addition,there have no negative effects to the blood routine,urine routine,stool routine,renal function,myocardial enzymes,blood lipids of Baishaoluoshi Decoction.ConclusionBaishaoluoshi Decoction can enhance the motor function,ameliorate the spasticity of upper limb,improve the daily living ability in post-stroke spasticity patients,what's more,it has a reliable security.Part 3 Experimental research ObjectiveTo observe the effect of Baishaoluoshi Decoction on post-stroke spasticity based on the synaptic plasticity and BDNF/Tr KB-KCC2 pathway.MethodsThe post-stroke spasticity model of rat was established by middle cerebral artery occlusion(MCAO).The rats were randomly divided into six groups as follows:high dose group;low dose group,baclofen group,model group,sham-operated group,and control group.The neurological deficit symptoms were evaluated by modified neurological deficit score(m NSS).Muscle tension were evaluated by Modified Ashworth score(MAS).Nissl's staining was used to observe the changes of Nissl bodies.Electron transmission microscope was used to observe the neurons and synaptic ultrastructure.The expression of synaptic plasticity-related protein BDNF, GAP43,p38,MAP2,and the GABA_A R?1,GABA_B1 R,GABA_B2 R in the brain tissue around the infarct were detected by Western blotting(WB).The expression of BDNF,Tr KB and KCC2 in the brain tissue around the infarct and the brainstem were detected by immunohistochemistry.Results1.The weight of the rats dropped sharply after ischemic stroke for about 10 days. Then the weight increased gradually,furthermore,the weight of rats in high dose and low dose group of Baishaoluoshi Decoction rebounded and increased faster than baclofen group and model group(P<0.01).2.MNSS show that there are statistically difference between the high dose,low dose group of Baishaoluoshi Decoction and model group,when 2 and 3 weeks after the intervention(P<0.01),but no statistically difference when 1 and 4 weeks after the intervention(P>0.05).3.MAS show that there are statistically difference between the high dose,low dose group of Baishaoluoshi Decoction and model group,when 3 and 4 weeks after the intervention(P<0.05),but no statistically difference when 1 and 2 weeks after the intervention(P>0.05).4.Nissl's staining shows that the Nissl body content of baclofen group and model group were reduced significantly compared with the sham-operated group,and control group(P<0.01).but in the high dose and low dose group of Baishaoluoshi Decoction,the content of Nissl body decreased relatively mild.5.The results of electron transmission microscope showed that there are a lot of damaged neurons and glial cells in the baclofen group and model group,the cytoplasm was concentrated,the mitochondria were swollen,the nuclear condensation and cleavage,the number of synapses decreased.But the damage was relatively light in the high dose and low dose group of Baishaoluoshi Decoction.6.The thickness of postsynaptic density(PSD)and the synaptic curvature increased significantly in the high dose and low dose group of Baishaoluoshi Decoction by electron transmission microscope(P<0.05),but the width of the synaptic gap have no difference(P>0.05).The expression of synaptic plasticity-related protein BDNF, GAP43,p38,MAP2 in the brain tissue around the infarct were raised remarkably by WB(P<0.01).7.The expression of BDNF,Tr KB and KCC2 in the brain tissue around the infarct and the brainstem were increased remarkably in the high dose and low dose group of Baishaoluoshi Decoction detected by immunohistochemistry(P<0.01).8.The expression of GABA_A R?1,GABA_B1 R,and GABA_B2 R in the brain tissue around the infarct have no difference by WB,intervened by high dose and low dose of Baishaoluoshi Decoction after 4 weeks(P>0.01).In all,the Baishaoluoshi Decoction can promote recovery of the body weight,improve the neurological deficits,relieve the spasticity of hemiplegic limbs and protect the damaged neurons and glial cells in ischemic stroke rats.Furthermore,these effects are related to the improvement of synaptic plasticity and the activation of BDNF/Tr KB-KCC2 pathway by Baishaoluoshi Decoction.ConclusionThe treatment of post-stroke spasticity by Baishaoluoshi Decoction may be related to improving the synaptic plasticity and regulating BDNF/Tr KB-KCC2 pathway.