| ObjectiveTo establish the cellular model of Alzheimer's disease induced by dissoluble Aβ25-35 segment neurotoxin.To explore the effect of Improved Wendan Prescription-contained(IWDP) cerebrospinal fluid and IWDP-contained serum on apoptosis and the expression of apoptosis associated protein bcl-2 and bax in cellular model of Alzheimer's disease applying the method of neurolymph pharmacological.And to analyze the possible mechanism of action in Improved Wendan Prescription's therapeutics to Alzheimer's disease further. MethodsAβ25-35 segment(5μmol/L) ageing treated beforehand was used to induce NG108-15 cells to establish the cellular model of Alzheimer's disease.The grouping were:blank group; model group; blank cerebrospinal fluid group;blank serum group; IWDP-contained cerebrospinal fluid group;IWDP-contained serum group.IWDP -contained cerebrospinal fluid and IWDP-contained serum was used to intervene in A β segment induced cellular model of Alzheimer's disease. To observe the cell morphous and growth state with inverted phase contrast microscope.Cell survival was assessed by counting MTT assay.Apoptosis level of NG108-15 cells was determined with flow cytometric analysis.The protein expression of bcl-2 and bax was examined by using SABC immunocytochemical method. Result 1 .The effect on cell morphous.The growth background of cells in blank group without Aβ25-35 treated was clear,while the proliferation in NG108-15 cells was prosperous. However,with the neurotoxic Aβ25-35 treated, we observed that the cell growth background was dirty, the dead cell and cell debris could be observed with microscopy,and the cell population was reduced,the cells growed clustering and gathering,most of the cells pericaryon became shrunken.The endochylema was dim,macula arised around the cells,cells with synapse reduced obviously. Compared with blank group, cells with synapse decreased significantly(p<0.05). Compared with model group, cells with synapse increased in IWDP-contained cerebrospinal fluid group and IWDP-contained serum group,the contrast was significant(p<0.05). 2.The effect on cytoactive.Cell survival rate was assessed by counting MTT assay,compared with blank group, the Cell survival rate in model group; blank cerebrospinal fluid group and blank serum group statistically decreased(p<0.05). Compared with model group,the optical density of IWDP-contained cerebrospinal fluid group and IWDP-contained serum group statistically increased(p<0.05). Compared with IWDP-contained serum group, the optical density of IWDP-contained cerebrospinal fluid group increased,but there were no significant differences between the two group ( p>0.05 ) . 3. The effect on Apoptosis. |
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