Protective Effects Of Gypenosides On Primary Culture Of Mesencephalic Dopaminergic Neurons In Rat Fetus

The degeneration and death of dopaminergic neuron in substantia nigra is themain pathological changes of Parkinson's disease (PD) , the current treatment forPD is still the way to control symptoms, there is no effective approach will alleviatethe progress of disease,while there will be more serious side effects of drugs affecttreatment. Gynostemma has already been widely used in traditional Chinese drugs,with no toxic side effects after long-term consumption. Studies have shown thatGynostemma extract various active ingredients as the neuroprotective effects ofantioxidants a lot. We hope that GPs in vitro by detecting the primary role inprotecting brain cells, screening the best drug concentration and time, to observe theanti-apoptotic and anti-oxidation, finally,we can provide experimental basis of GPsfor clinical treatment of PD.Objective: In order to use gypenosides(GPs) for clinic therapy of PD, theprotective effect of GPs on the primarily cultured neuronal cells from embryonic ratmesencephalon was observed,and the best concentration and time were screened out.Methods: Primarily cultured cells of embryonic rat mesencephalon were divided into the control group, the positive control group(NGF), the experimentalgroups(GPs).The final concentration of GPs: 100μg/mL, 200μg/mL, 400μg/mL,800μg/mL, and after the action of every 6h,12h,24h,48h,72h and 96 hours, the cellswere observed by inverted microscope, and the activity was detected by MTTassay;the results were analysed to identify the best concentration and time. Tyrosinehydroxylase(TH) immunocytochemical staining was used to compare the numberand figure of positive neurons of each group. Disposed with MPTP,the antiapoptosiseffect of GPs were analyzed through Flow cytometry, and the antioxidationeffect was observed by inducible nitric oxide synthase(iNOS)immunocytochemical staining. Results: According to MTT measured by the valueof OD , GPs 400μg/mL acting 48 hours got the value of 1.006±0.062, comparedwith NGF acting 48h group by no statistical difference(P>0.05), while comparedwith others by significant difference(P <0.05).At this state the numbers of apoptosiswere the least, and the express level of iNOS was decreased. Conclusion: At theconcentration of 400μg/mL acting 48 hours, GPs has protective effect againstMPTP-induced danmage to mesencephalic cultured cells from rat embryos.