The Effect Of Electro-acupuncture Combined With BMSCs Transplantation On Learning And Memory Ability Of Rats Subjected To Intracerebral Hemorrhage And Its Mechanism

Objective:To observe the effect of electro-acupuncture(Ea) combined with bone marrow mesenchymal stem cells (BMSCs) transplantation on learning and memory ability of rats following intracerebral hemorrhage (ICH), and study its mechanism by investigating the levels of SOD, MDA and AchE in the serum and the expressions of SYP, GAP-43and PSD-95protein in the hippocampus. Methods:(1)The BMSCs were recovered, amplified, induced and collected to use.(2) Adult SD rats were trained by Morris water maze four days, four times a day. On the fifth day, rats with less than40seconds of escape incubation period were randomly divided into BMSCs group, Ea group, Ea-BMSCs group, NS group, FO group and Normal group.(3) In the previous four groups, the ICH rat models were established by injecting collagenase and heparin into caudate nucleus. Three days after operation, the4groups of rats were treated by BMSCs transplantion, Ea, Ea combined with BMSCs transplantion and injection of normal saline respectively. Each group was subdivided into1d,3d,5d,7d,14d five subgroups based on refeeding time. Ea stimulated rat by the Dazhui and Baihui, once a day. FO group was only inserted the needle without the injection of collagenase and heparin, and didn t accept any other treatment. Normal group hadn’t any interventions.(4)In the corresponding time, rats of each group were tested by Morris water maze, then the serum and brain were collected.(5)We detected the levels of AchE, SOD and MDA in serum, and expressions of the SYP, GAP-43and PSD-95protein by immunohistochemistry. Results:(1) There is no significant difference (P>0.05) between the groups before modeling by water maze test. After the treatments on ICH, the escape latency, the frequency of passing through platform, the platform quadrant time and distance percentage which reflected the learning and memory abilities were improved with the passage of time by water maze test. The learning and memory ability of Ea group was better than the other groups on the1d after treatmemt(P<0.05), and that of NS group was worse than the other groups (except BMSCs group on3d) on the3、5、7、14d (P<0.05), and that of Ea-BMSCs group was better than Ea group and BMSCs group (P<0.05).(2)In ICH groups, the content of AchE began to decline on1d, and reduced to the lowest level on14d. The content of AchE in the NS group was higher than that of the other treatment groups on5、7、14d (P<0.05); The content of AchE in the Ea-BMSCs group was higher than that of the Ea group on5d(P<0.05), but lower than that of the Ea group and the BMSCs group on7、14d (P<0.05).(3)The content of SOD in serum of ICH groups began to decline on1d, and reduced to the lowest level on3d, then increased rapidly and reached the peak on14d. The content of SOD in the Ea group was higher than that of the other treatment groups on3d(P<0.05). The content of SOD in the NS group was lower than that of the other treatment groups on5、7、14d(P<0.05). The content of SOD in the Ea-BMSCs group was higher than that of the Ea group and BMSCs group on7、14d(P<0.05). The content of SOD in the BMSCs group was lower than that of the Ea group on5d(P<0.05).(4)The content of MDA in the serum of ICH groups began to rise on1d and reached the peak on3d, then decreased and reached the lowest level on14d. The content of MDA in the Ea group was lower than that of the other treatment groups on3d(P<0.05). On5、7、14d, the content of MDA in the NS group was higher than that of the other treatment groups (P<0.05), the content of MDA in the Ea-BMSCs group was lower than that of the Ea group and BMSCs group (P<0.05). The content of MDA in the BMSCs group was higher than that of the Ea group on5d(P<0.05).(5)In ICH groups, the expression levels of SYP、PSD-95began to rise on1d, Ea-BMSCs group and BMSCs group increase continully and reached the peak level on14d. Ea group and NS group reached the peak level on7d, and then decreased. The expression levels of GAP-43began to rise on1d and reached the peak level on7d, then decreased.(6) On5、7、14d, the expression of SYP, GAP-43in NS group was lower than the other treatment groups (P<0.05), the expressions of SYP, GAP-43in Ea-BMSCs group were higher than Ea group and BMSCs group (P<0.05).(7) On3、5d, the expression of PSD-95in Ea group was higher than other treatment groups (P<0.05). On5、7、14d, the expression of PSD-95in NS group was lower than other treatment groups (P<0.05).On7d、14d, the expression of PSD-95in Ea-BMSCs group was higher than Ea group and BMSCs group (P<0.05).(8)After14days of treatment, SOD, SYP, PSD-95, MDA, AchE, GAP-43, learning and memory ability of each treatment group compared with the FO groups and the Normal group, there are still significant differences (P<0.05). Conclusion:(1)Compared with the Ea treatment or BMSCs transplantation, Ea combined with BMSCs transplantation can significantly improve the ability of learning and memory in ICH rat, which may be associated with decrease of AchE content in serum and increase expression of SYP, PSD-95and GAP-43Protein in the hippocampus.(2)Treatment of Ea combined with BMSCs transplantation on ICH rat can increase SOD activity in serum significantly and reduce the production of MDA, thereby reduce the damage to the body by free radical.