Comparative study of standardized Centella asiatica and combination of levodopa and benzerazid HCl on Parkinson′s model zebrafish

OBJECTIVE To investigate the motility,dopamine,serotonin and pro-BDNF level of Centella asiatica(CA)comparable to the combination of levodopa and benzerazid HCL on zebrafish model Parkinson′s.METHODS Rotenone 5μg·L-1 induced to adult zebrafish(8months)for 28 dto made zebrafish model Parkinson′s.CA concentration were used 5 and 10μg·mL-1.The combination of levodopa 100 g and 25 mg benzerazid(L-DOPA agonist)was given at 1 μg·mL-1 at the same time to rotenone,and CA as well.Motility assessment conducted for every 7dstarted on day 0until day 28.Five fish were subjected in the 2L tank(25cm×16.5cm×12cm).Three vertical lines were drawn on the tank at equal distances,dividing the tank into four zones(the length of each zone was 6.25cm).Locomotor activity was measured for 5min by counting the number of lines that adult zebrafish crossed.Fish were sacrificed by decapitating on ice water.Dopamine level measured from whole brain by ELISA,serotonin and pro-BDNF by immunoreactivity at substantia nigra.RESULTS The locomotor activity of rotenone treated fish were significantly decrease starts at 7 dcompared to control group.Interestingly on 10μg·mL-1 group there are increasing motility start at day 7 and slightly decrease until day 28,but on L-DOPA group increasing motility at day 7 followed by decreasing motility significantly(P<0.05)until day 28.Dopamine level of rotenone group decreased compared to control group,CA 10μg·mL-1 and L-DOPA group increased significantly to rotenone group which CAμg·mL-1 higher than L-DOPA group.Serotonin and pro-BDNF level had the same profile.Its significantly increased on rotenone group compared to control group.Serotonin on group with CA administration significantly decreased compared to rotenone group,while rotenone plus CA 10μg·mL-1 had no significant differet to L-DOPA group(P>0.05).Pro-BDNF on rotenone with CA 5μg·mL-1 had no significant difference,while compare to rotenone with CA 10-1 μg·mLgroup and L-DOPA group were significantly different(P<0.05).CONCLUSION Rotenone produces reliable Parkinson′s zebrafish model by decreasing motility and dopamine level.CA was more stable,increasing motility than L-DOPA administration.CA and L-DOPA increased dopamine level,but decreasing serotonin and pro-BDNF.