The Change Of The Ethology And Cholinergic System Of The Hippocampus Formation By Intravenous Administration Of Bone Marrow Mesenchymal Stem Cells With Mannitol Among Vascular Dementia Rats

PARTⅠEstablishing Vascular Dementia Model In Rats【Objective】To build a model of vascular dementia(VD) in rats.【Methods】The healthy matured-male SD rats were divided randomly into two group .○1 sham group: only cutting the skin of neck ,no ligation.○2VD model group: The models of VD rats were established through permanent ligation of bilateral common carotid arteries at an interval of 3 days for each artery. Four week after ligation , The learning and memory ability of two group rats were tested by Morris water maze.【Results】The survival rate of VD rates reached up to 52.17% ,four week after ligation. Compared with sham group, The learning and memory ability of VD model group rats were receded obviously(P<0.05)【Conclusions】The improved permanent bilateral common carotid artery occlusion is a better method to establish the VD model. PARTⅡIsolation,Cultivation And Identification Of Rat Bone Marrow Mesenchymal Stem Cells【Objective】To culture rat bone marrow mesenchymal stem cells (BMSCs) fitting the requirement of experimentation for further study.【Method】Adopt whole bone marrow adherent cultivation to culture rat BMSCs. When cells covered with plastic bottom 80% -90%, digest them with 0.25% trypsin in 120S and then passage the cells.Observate the morphology and characteristics of cultured cells by inverted phase contrast microscope ,identify the surface marker identification of CD44,CD90 and CD34 of cells by flow cytometry. And,osteogenic differentiation of BMSCs to identify BMSCs after cell amplification.【Results】BMSCs growed with adherence and exhibited a spindle-shaped and spindle shape, After passaged into the third generation BMSCs'morphology became similar. Cells cultured by whole bone marrow adherent cultivation showed CD44(89.4%),CD90(99.2%) and CD34(1.82%). BMSCs have multiple differentiation potential, can be induced into osteoblasts. Alkaline phosphatase staining was positive,Alizarin red staining showed the calcium mineralization positive.【Conclusion】Whole bone marrow adherent cultivation can successfully construct a rat BMSCs isolation and culture system in vitro, and culture BMSCs with high purity, energy intensity and stable characters. PARTⅢThe change of the ethology and cholinergic system of the hippocampus formation by intravenous administration of bone marrow mesenchymal stem cells with mannitol among vascular dementia rats【Objective】To explore The change of the ethology and cholinergic system of the hippocampus formation by intravenous administration of bone marrow mesenchymal stem cells (BMSCs)with mannitol among vascular dementia(VD) rats.【Methods】The BMSCs of rat were isolated and cultured by whole bone marrow adherence screening method. The models of VD rats were established through permanent ligation of bilateral common carotid arteries at an interval of 3 days for each artery.The healthy matured-male SD rats were divided randomly into four group.①Mannitol with BMSCs group: intravenous injection of mannitol at a dose of 1.5 g/kg followed 10-30 minutes later by intravenous injection of 1 million BMSCs in 1ml PBS at the fourth week after ligation.②BMSCs group: intravenous injection of 1 million BMSCs in 1ml PBS at the same time.○3 PBS control group: intravenous injection of only 1ml PBS.○4sham group:no ligation and no intravenous injection.the ethology and The activity of choline acetyltransferasecholine (ChAT) and acetycholinesterase (AChE) were tested at the fourth week after injection.【Results】The activity of ChAT and AchE significantly improved in the hippocampus of rats in Mannitol with BMSCs group compared with BMSCs group and PBS control group(P<0.05),so did the ethology.The same significant improve in BMSCs group compared with PBS control group(P<0.05).【Conclusion】BMSCs improve the ethology and cholinergic system activity of the hippocampus formation of VD rats.The beneficial effects of intravenous injection of BMSCs are amplified with intravenous injection of mannitol.