Shenmayizhi Prescription Alleviates Neurovascular Units Deficits In Vascular Dementia Rats

ObjectiveTo observe the effect of herbal compound Shenma Yizhi prescription on the neurovascular unit(NVU)of vascular dementia(VD)rat model including the blood-brain barrier,neurons and glial cells,thus exploring the mechanism of Senma Yizhi Decoction in the treatment of vascular dementia.MethodsGrouping,modeling and administration70 male Wistar rats(aged lOweeks,weight 300±40g)were randomly divided into 7 groups with ten in each.The groups are as follows:controlgroup,sham operation group,MID group,donepezil group,low dosage traditional Chinese medicine(TCM)group,medium dosage TCM group and high dosage TCM group.Rats in the model group,donepezil group and the TCM groups were treated with the method of autologous micro-thrombus embolization.Sham-operated rats were injected with 0.3ml physiologic saline.After operation,the donepezil group was given 0.5mg/kg intra-gastric administration of drugs,low dosage group 3.3mg/kg,medium dosage group 6.6mg/kg,and high dosage group 16.5mg/kg,and the sham operation group and model group wereadministrated equivalent distilled water.The intra-gastric administration were taken once a day for 4 weeks.Chapter 1Behavioral tests and morphology of hippocampus of VD rats1 MethodsMorris water maze was used to test the learning and memory ability of rats in each group.Then the whole brains of rats were cut and fixed,paraffin embedded,sectioned,and stained with HE or with Nissl's method.2 ResultsCompared with sham operation group,the escape latency of the model group was prolonged every day;the passing platform times(PPT)were significantly reduced;The fourth quadrant time was shortened;and the fourth quadrant distance was reduced.The difference was statistically significant(P<0.01).Compared with model group,the escape latency of the donepezil group,Shenma Yizhi prescription medium dosage group and high dosage group were shortened every day,PPT and the fourth quadrant distanceof the donepezil group and Shenma Yizhi prescription high dosage group were significantly increased,The difference was statistically significant(P<0.01).Chapter 2Shenma Yizhi prescription promoted NVU angiogenesis of the VD rats.1 MethodsWestern blot was used to detect HIF1 ? and Notch content of the rats'brain tissue,ELISA to detect VEFG content,RT-PCR to detect VEGmRNA content,and immunohistochemistry to detect the expression of CD34.2 Results2.1 HIF1? and Notch expressionsCompared with model group,the HIF1 ?,VEGF and Notch protein expression of Shenma Yizhi prescription medium dosage group and high dosage group were increased in a dose-dependent manner.The difference was statistically significant(P<0.01).2.2 VEGFmRNA contentCompared with model group,the VEGFmRNA protein content expression of Shenma Yizhi prescription high dosage group were increased.The difference was statistically significant(P<0.01).3 conclusions3.1 Shenma Yizhi prescription can enhance the HIF1?,VEGF and Notch protein expression in a dose-dependent manner.3.2 Shenma Yizhi prescription can promote the VEGFmRNA secretion.Chapter 3 Shenma Yizhi prescription inhibited neuron apotosis and glial cell activatianin NVU of the VD rats1 MethodsWestern blot was used to detect Nrf2,HO-1 and caspase3 expression of the rats' brain tissue;RT-PCR to detect Nrf2mRNA and HO-1 mRNA expression;colorimetry to detect the activity of CAT,GSH,GSH-PX and SOD and MDA content;Elisa to detect IL-1 and TNF-? expression;and immunohistochemistry to detect iba-1,GFAP and NEUN.2 ResultsCompared with model group,the Nrf2 and HO-1 protein expression of Shenma Yizhi prescription medium dosage group and high dosage group were increased;the Nrf2mRNA expression were increased,the HO-1 mRNA expression of Shenma Yizhi prescription high dosage group were increased;In Shenma Yizhi prescription medium dosage group and high dosage group the caspase3 protein expression were decreased;the activity of CAT,GSH,GSH-PX and SOD were increased;MDA content were decreased;IL-1 and TNF-? protein expression were decreasedinShenma Yizhi prescription high dosage group.The difference was statistically significant(P<0.01,P<0.05).Chapter 4Shenma Yizhi prescription alleviated BBB impairments of the VD rats1 MethodsWestern blot was used to detect CX43 and AQP4 protein expression of the rats' brain tissue2 ResultsCompared with model group,the CX43 and AQP4 protein expression of medium dosage group and high dosage group were decreased.The difference was statistically significant(P<0.01).