| Objective: To investigate the effects of combination of rhG-CSF and VEGF on the cognition disorders ,neural stem cell regensis and antiapoptosis of senescence accelerated mouse P10Method: 90 SAM-P10 were selected and divided randomly into control group, rhG-CSF group, rhG-CSF and VEGF combined group. Mice in rhG-CSF group were given rhG-CSF 100 ug/(kg·d) subcutaeneuslly one time per day for 7 days, while mice in the rhG-CSF and VEGF combined group were first given VEGF 50 ug/(kg·d) intraperitoneally one time per day for 3 days and then rhG-CSF 50 ug/(kgd) subcutaeneuslly one time per day for 7 days. Control group were given normal saline. And then all the mice were given Brdu 50 ug/(kg·d) intraperitoneally one time per day for 7 days, Morris water maze test was done before administration and on the 7th,14th,28th,and 56th day after administration , record the escaping latency and the percentage of swimming distance in the target quadrant of each mouse. After each test,execute the mice,make paraffin slice up and then obeserve the expression of nestin,Brdu and caspase-3 by immunohistochemistry method respectively. Take photoes of all the pictures and count the immunity positive cells. SPSS 11.0 for Windows was used to analyze the data. All the data were expressed as means±S.D. (x ± s) and analyzed by ONE-WAY ANOVA P<0.05 was taken as the statistical standard.Result:1. Morris water maze test result1.1 Place navigation: The escaping latency of control group increased obviously on the 14th,28th,56th day(P<0.05); In the rhG-CSF group, there was no significant difference between each time point(P>0.05) ,while the escaping latency was shorter than the control group on the 14th day(P<0.05); In the combination group, the escaping latency was shorter than before administration and was also shorter than the control group on the 14th,28th,56th day(P<0.05),moreover,it was shorter than the rhG-CSF group on the 56th day(P<0.05).1.2 Spatial probe test result In the control group,the percentage of swimming distances in the target quadrant increased on the 28th,56th day than before administration;In the rhG-CSF group.it was longer than the control group on the 14th day(P<0.05);In the combined group,it was longer than before administation on the 28th ,56th day and also longer than the control group on the 14th ,28th ,56th day,moreover,it was longer than the rhG-CSF group on the 56th day (P<0.05).2. Observe the expression of nestin around the third ventricle by immunohistochemical methodThere are a few nestin immunity positive cells aroud the third ventricle at each time point in the control group,In the rhG-CSF group .there are nestin immunity positive cells aroud the third ventricle at each time point ,with no significant difference than before administration(P>0.05),while there are more than the control group on the 7th, 14th day(P<0.05 .In the combined group,it increased than before administration on the 56th day and also more than the control group on the 7th ,14th,28th day,while increased on the 56th day than the rhG-CSF group(P<0.05)3. Observe the expression of Brdu around the third ventricle by immunohistochemical methodThere are a few Brdu immunity positive cells aroud the third ventricle at the 7th, 14th day in the control group,In the rhG-CSF group ,there are nestin immunity positive cells aroud the third ventricle at each time point ,with no significant difference than before administration(P>0.05),.In the combined group,it increased than before administration on the 7th,14th day(P<0.05 ) and also more than the control group on the 7th ,14th,28th day,while increased on the 7th day than the rhG-CSF group(P<0.05)4. Observe the expression of caspase-3 in hippocampus by immunohistochemical method There are immunity caspase-3 positive cells in hippocampus at each time point and increased than before administration at the 28th ,56th day in the control group,In the rhG-CSF group ,there are nestin immunity positive cells in hippocampus at each time point ,with no significant difference than before administration(P>0.05),and fewer than the control group on the 7th ,14th day. In the combined group,it decreased than before administration on the 56th day(P<0.05 ) and also fewer than the control group on the 7th ,14th day,while decreased on the 56th day than the rhG-CSF group(P<0.05)Conclusion:1.rhG-CSF can improve the cognition disorders of SAM-P10,and delay the decline of its learning and memory ability to some extent, rhG-CSF combined with VEGF is able to improve the cognition disorders of SAM-P10obvisously with long term effect.2. rhG-CSF can increase the nestin immunity positive cells of glia cell in the third ventricle of SAM-P10 to some extent, The effect is more obvious in in the rhG-CSF combined with VEGF group,they can promote the compensatory repair of damaged neuron and participate the formation of nerve connection.3. rhG-CSF can increase the Brdu immunity positive cells in the third ventricle of SAM-P10 in the commitment, The effect is more obvious in in the rhG-CSF combined with VEGF group,they can promote the in situ proliferation of endogenous NSC.4. rhG-CSF can decrease the caspase-3 immunity positive cells in the hippocampus of SAM-P10 to some extent, The effect is more obvious in in the rhG-CSF combined with VEGF group and they can inhibit the apoptosis of the hippocampal neuron...
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