Improving Effect Of Soybean Phosphatidylcholine On The Motor And Cognitive Function Impaired Cerebral Palsy Of Neonatal Rats

AIM: the study was to investigate the effect of soybean phosphatidylcholine (PC) on the motor and cognitive function impaired cerebral palsy of neonatal rats . The immunoreactivity of brain derived SYN and ChAT were performed by immunohistochemical method in hippocampus formation. The potential mechanism of moving and cognizing ability in normal feeding young rats and CM+CP young rats was revealed further.METHODS: 14 days Wistar rats (25 g to 30 g) sex regardless were randomly divided into 4 groups. Experimental condition was coordinated, 20+2°C, 60%—70% humidity. 12 h periods of light and dark (light: 7: 00 am-7: 00 pm) . (1) cage control group(CC group, n=24), rats were housed individually in cage, fed by standard laboratory chow and tap water for 20 days, 10ml/kg tap water was administered by gastrogavage once per day;(2) false operating group(SO group, n=24), rats sham operation then housed individually in cage, fed by standard laboratory chow and tap water for 20 days, lOml/kg tap water was administered by gastogavage once per day;(3) cerebral palsy group(CM group, n=24),rats were made into model of cerebral palsy ,housed individually in cage, fed by standard laboratory chow and tap water for 20days, 10 ml/kg tap water was administered by gastogavage once per day;(4) cerebral palsy madel treated with PC group (CM+PC group, n=24), rats were made into model of cerebral palsy then housed individually in cage and fed by PC for 20 days. Soybean phosphatidylcholine(PC) 400mg/kg/d were dissolved with tap water and administered by gastogavage once per day to rats. From the 2nd day, use the beam walking test system,footprint pattern (gait) test system respectively to test, analyze and evaluate the curative effect on obstacles in moving and coordinating, moving and balancing as well as cognizing ability.rats were tested every other day for 10 days in total. From the 11th day, rats were tested respectively with morris water maze every 3 sequential days. Two trials were conducted from each starting point. The data of swimming distance (D), latency(L), memory score (MS), velocity(V) for searching submerged platform were recorded and analyzed by behavioral analyses system.rats were executed in 3 batches after each experiment. 6 rats were executed in 5 days, another 6 rats in 10 days, and the left 6 rats in 20 days. The young rats were perfused by of a fixative solution through intra-aortic, then the immunoreactivity of the positive cells in brain derived SYN and ChAT were performed by immunohistochemicalmethod in hippocampus formation. Hippocampus formation of rats was dissected for revealing immunoreactivity of SYN and ChAT by using SABC immunohistochemical method.RESULTS: 1. results of the learning and memory ability tested by the morris water maze(1)CM+PC group, CM group compared with SO group, the latency(L) for searching submerged platform increased after making the model for 11 to 30 days, and there were significant differences(P<0.05), after making the model for 17 to 19 days, there was no difference in CM+PC group compared with SO group (P>0.05 ), but there were significant differences in CM group compared with SO group (PO.01) . and significant differences existed in CM+PC group compared with CM group (P<0.05 ). it showed the PC had effect in improving cognizing ability;SO group vs CC group (P>0.05 ) , there was no difference between SO group and CC group.(2)there was no significant difference in 4 groups comparing to the swimming belocity (.P>0.05), this showed making the model didn't decrease rats' physical strength significantly.(3)CM+PC group, CM group compared with SO group, after making the model for 11 to 13 days, the swimming distance for searching submerged platform increased with significant difference ( P<0.05 );after making the model for 17 to 19 days, there was no difference in CM+PC groupcompared with SO group, but there was significant difference in CM group compared with SO group (P<0.01);and significant difference also existed in CM+PC group compared with CM group (P<0.05) , it showed the PC had effect in improving cognizing ability;SO group vs CC group(P>0.05), there was no difference between SO group and CC group. 2. Results of footprint pattern testing(1) Comparing the left side step length of CM+PC group;CM group to SO group, there was difference compared with the contrasted groups after making the model for 1 to 9 days, CM+PC group, CM group vs SO group (PO.05), but there was no difference after 11 days, it maybe because of the short-term symptoms caused by the opposite side palsy;CM+PC group vs CM group(P>0.05), CC group vs SO group(P>0.05), there was no difference between CM+PC group and CM group, there was also no difference in the left side step length between SO group and CC group.(2) comparing the right side step length of CM+PC group, CM group to SO group(P<0.05), cereral palsy resulted in the growth of the right side step length;CM+PC group vs CM group(P<0.05), PC can improve this moving obstacles;CC group vs SO group(P>0.05), there was no difference in the right side step length between SO group and CC group.(3) there was no difference in the forelimb's distances among the 4 groups (P>0.05) .(4) comparing the hind legs' distances of CM+PC group , CM group to SOgroup (PO.05) , cerebral palsy resulted in the growth of the hind legs' distances, and there was significant difference compared with the contrasted group;CM+PC group vs CM group (P<0.05) , the PC had significant curative effects;SO group vs CC group (P>0.05), there was no difference in the hind legs' distances between SO group and CC group .(5) there was no difference in the superposed comparison of the left side's forelimbs and hind legs (/*>(). 05 ) .(6) comparing the right side's forelimbs and hind legs of CM+PC group. CM group to SO group (P<0.05) , the superposed odds of the right side's forelimbs and hind legs decreased while the distance between the two core points increased because of cerebral palsy, there was significant difference compared with the contrasted groups;CM+PC group vs CM group(.P<0.05 ) , PC had effects in impriving this moving obstacles;CC group vs SO group (.P>0.05) , there was no difference in the superposed comparison of the rifht side's forelimbs and hind legs.3. results of beam walking(1) from the 1st day to 19* day, there was significant difference among SO group, CC group and CM group after comparison (/><0.05, P<0.0\) . it showed the cerebral palsy model had been set up successfully, there was significant difference in moving coordination and balance. Soybean phosphatidylcholine (PC) were dissolved with tap water and administered by gastrogavage for 1 to 19 days, there was significant difference inCM+PC group compared with CM group (P<0.05 , P<0.01);it showed the PC had effects in improving the moving coordination and balance of cerebral palsy model, and the cerebral palsy model was tending to be normal day by day.4. results of immunobistochemical testing(1) the positive immunoreactive labeled SYN and ChAT in the hippocampus formation increased in CM+PC group compared with CM group and SO group (P<0.05 ) , while there was no changes compared with SO group and CC group (P>0.05) .CONCLUSIONS: (l)long-term intake of phosphatidylcholin can improve the moving, cognizing ability of cerebral palsy young rats. (2)phosphatidylcholin protects the central nervous system of cerebral palsy young rats, via the potential mechanism of providing extra-phosphatidylcholine for synthesizing acetylcholine and enchance SYN and ChAT activity in hippocampus.