| Objective:To explore effects of acupuncture on the expression level of GAP-43 in the contralateral cortical motor area of rats with cerebral ischemia and its relationship with the rehabilitation of rats' neurological function recovery.Methods:One hundred SPF male rats were randomly divided into 5 groups: the sham-operation group,the model group,the unaffected-side acupuncture group,the affected-side acupuncture group and the bilateral acupuncture group with 20 rats in each group.All the rats are raised in the same condition before any treatment.The sham-operation group and the model group were treated with nothing and practice acupuncture on the rats of the other groups.The affected-side acupuncture group was treated with acupuncturing “Jianyu” “Quchi” “Hegu” “Sanyinjiao” “Zusanli” “Huantiao” on the affected-side.The unaffected-side acupuncture group was treated with acupuncturing “Jianyu” “Quchi” “Hegu” “Sanyinjiao” “Zusanli” “Huantiao” on the unaffected-side.The bilateral acupuncture group was treated with acupuncturing “Jianyu” “Quchi” “Hegu” “Sanyinjiao” “Zusanli” “Huantiao” on both sides.After inserted the needle retained 30 minutes and during which manipulated the needles evry 10 minutes.Treatments were given once a day and 6 days a week and one week is a course with one day off between two courses.Obtain the rats' neurological function score when it was 24 hours after the first treatment,and on the 7th day,the 14 th day,the 21 st day and the 28 th day since started.On the other hand behead 5 rats from each group every 7days.Immunofluorescence stain the GAP-43 of the embedded section made of the contralateral cortical motor area(from 4.5mm before the bregma to 0.5mm after the bregma)and observe the GAP-43 expression level.Results: 1.The rats' neurological function score of each group The model group and the acupuncture group's Zea-Longa score and the score obtained 24 hours after the first treatment compared with the sham-operation group and the difference was statistically significant(P < 0.05).The model group and acupuncture group's Zea-Longa score compared with the score obtained 24 hours after the first treatment and there was no statistically significant difference(P > 0.05),which is comparable.The rats' comprehensive score of acupuncture group were higher than the model group(P < 0.05).The scores of the unaffected-side acupuncture group,the affected-side acupuncture group and the bilateral acupuncture group on the 7th,14 th,21st and the 28 th day were statistically different(P<0.05).On the 7th,14 th,21st and 28 th day compared between the unaffected-side acupuncture group,the affected-side acupuncture group,there was no statistically significant difference(P > 0.05)but the difference between the bilateral acupuncture group and the affected-side acupuncture group is statistically significant.The differences within each groups on the 7th,14 th,21st and the 28 th day were statistically remarkable(P<0.5).The differences between the 21 st day and the 28 th day were not statistically significant within both the affected-side acupuncture group and the unaffected-side acupuncture group(P>0.5),but it was quite the opposite within the bilateral acupuncture group(P<0.05).2.The GAP-43 expression level of experimental rats ' the contralateral cortical motor area in each groups.The GAP-43 red fluorescence signal of the sham-operation group was weak(±),and a few scattered signal on the cell membrane can be seen(as shown in the figure 2).The GAP-43 red fluorescence signal of the model group were scattered on a few cell membrane which was discontinuous and weakly positive(±)(as shown in the figure 3).The GAP-43 red fluorescence signal of the acupuncture groups can be observed all over the cell membrane which is increasingly strong.In the bilateral acupuncture group the GAP-43 positive cells were ellipse and full,with one or two outgrowth appearing,which is the typical neurons form(as shown in the figure 4).No typical neurons form was observed in the affected-side acupuncture group,the unaffected-side acupuncture group(as shown in the figure 8 and 12).On the 14 th day the GAP-43 red fluorescence signal is the strongest and there were more positive cells than on the 7th day and the cells were ellipse and continuously closely arranged(as shown in the figure 5,9 and 13).On the 21 st day,the GAP-43 red fluorescence signal is continuously expressed on the cell membrane but weakened.And in the affected-side acupuncture group,the unaffected-side acupuncture group the typical neurons disappeared(as shown in the figure 10 and 14),but some can be observed in the bilateral acupuncture group(as shown in the figure 6).On the 28 th day,the GAP-43 red fluorescence signal kept getting weaker to the level similar to the sham-operation group and only a few scattered signal could be observed on the cell membrane.And the GAP-43 positive cells disappeared in the affected-side acupuncture group and the unaffected-side acupuncture group(as shown in the figure 11 and 15),and GAP-43 positive cells scattered like stars in the bilateral acupuncture group(as shown in the figure 7).3.The GAP-43 positive immune cells' relative ratios from the contralateral cortical motor area of experimental rats in each group.According to the ANOVA analysis,the ratio of GAP-43 positive cells to all the cells among each groups were statistically different(F=8.8311,P<0.05),which illustrated that in each group the GAP-43 expression level in the cells from the contralateral cortical motor area of experimental rats are different.The number of GAP-43 positive immune cells in the acupuncture groups were much higher than they were in the sham-operation group,and there was not much difference between the affected-side acupuncture group,the unaffected-side acupuncture group,and the bilateral acupuncture group showed the highest number.Conclusions: 1.Acupuncture treatment can enhance the neurological function recovery of rats with cerebral ischemia.Compared with the model group,the neurological function had been improved obviously in the affected-side acupuncture group,the unaffected-side acupuncture group and the bilateral acupuncture group among which the bilateral acupuncture group worked the best.So bilateral acupuncture group had a certain advantage in rat's neurological functional recovery.2.Acupuncture treatment could promote the GAP–43 expression in the contralateral motor area of rats with cerebral ischemia.Compared with the model group,the GAP–43 expression level was much higher in the affected-side acupuncture group,the unaffected-side acupuncture group and the bilateral acupuncture group among which the bilateral acupuncture group was the most effective and the GAP–43 expression time was prolonged.As a result,the bilateral acupuncture group has a certain advantage in promoting the expression level of GAP-43 in the contralateral cortical motor area of rats with cerebral ischemia. |
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