| Parkinson's disease(PD) is a degenerative disease in the central nervous system, the clinical manifestations of PD usually like chronic progressive movement slowly, tremor and muscle rigidity, but the general course may have no natural ease. With the aging getting more and more serious, the number of the onset of Parkinson's disease, the incidence of the disease had presenting a growth trend. Because the etiology of Parkinson's disease is unclear, and it is just for the prevention but the effective drug for the treatment of Parkinson's disease has not been found particularly, so the discovery of active substance and its mechanism for anti-Parkinson's disease has been the hot topics in the study of the medical profession. Group committed to the research of Buddleja lindleyana Fort. for a long term, and found triterpene compounds of Buddleja lindleyana Fort. has certain nerve protective effect, showing a certain resistance to PD activity in the early study. Therefore, in this paper, on the basis of previous research discovers, we established the model of Parkinson's disease through the wild-type caenorhabditis elegans(C.elegans) for screening anti-PD active compounds from Buddleja lindleyana Fort., and study the compound effect and action mechanism on Parkinson's disease, in order to provide a new research idea for the study of traditional Chinese medicine(TCM) modernization, for new drug research and development to provide a large number of sources.Objective: To established a new type model of Parkinson's disease through C. elegans, and used the model to selecte the active ingredient of Parkinson's disease from Buddleja lindleyana Fort.; Q tof MS technology was applied to the active ingredient buddleia fruit for preliminary studing.Methods:(1) Adoptted the way of drug induced model, and then set up the concentration gradient of MPP+ 0.6~1.6 m M(1-methyl-4-phenyl-pyridine salt ions, as active metabolites of MPTP(1-methyl-4-phenyl-5-four hydrogen pyridine)), type of wild C. elegans injuries respectively, then investigating the fertilized egg number,number of offspring, head oscillating frequency, unilateral gonad arm oocytes, mortality rates five indexes; At the same time, we had used transgenetically nematodes(such as: dopaminergic neurons cat-2, interneurons TTX-3, chemical sensory neurons STR-1, other sensory neurons TPH-1)(i.e., head neurons are markers of GFP nematodes), from the Angle of molecular biology, further verified the MPP+ whether it had certain effects on dopaminergic neurons, the positive control drugs(effective drug for anti-Parkinson's diease: levodopa, Lisuride, Apomorphine) had been used for verifing the above results and optimizing the best concentration of building;(2) Used the model which had been successfully established for screening and tracking active ingredient of anti-Parkinson's disease from buddleia fruit, and shown in the extracted parts;(3) Used analysis technology---Q-tof-MS to ended up with the activity of anti- Parkinson's disease for the component identifying and primary analysising of extracted parts.Results:(1) We have proved that the MPP+ can be used as a drug for induced damaging to establishing PD model on caenorhabditis elegans, and 1 m M MPP+ can be an optimal concentration for the model establishing;(2) We had tracked the activity compounds in 8 samples which reflecting different degree activities of anti-Parkinson's disease by the experiments;(3) As the Q tof MS analysis said, sample S4 of the communist had been identified 11 compounds, respectively is: isomartynoside, cranioside A, 4-O-acetylmartynoside, daucosterol, Echinatic acid,buddlintriterpene B, et al; Sample S5 of the communist had been identified 9 compounds, which including: cranioside A, desrhamnoverbascosapain, 13,28-epoxy-3?,23-dihydroxy-11-oleanene, et al; Samples S6 has respectively identified five flavonoids, 2 styrene acrylic phenolic glycosides, four triterpene, the three types play a role commonnly; Classics appraisal, sample S10 mainly contained triterpene compounds, such as, buddlintriterpene A, buddlintriterpene B, mimengoside A, mimengoside I and so on; Samples S11 had been identified one styrene acrylic phenolic glycosides, three terpenoids and one sterols; There were five triterpene compositions in the sample S13; In the samples S14 had been identified three terpenoids and one sterols; Sample S16 was mainly the triterpenoid saponins compounds make roles in anti-PD.Conclusion: The new model of animal — C. elegans can be used to pharmacology researching and drug high-throughput screening researching as an experimental model; There has been active ingredient for Parkinson's disease resistancing in Buddleja lindleyana Fort., but the specific mechanism remains to be further studied. At the present stage, the existed active ingredient of anti-Parkinson's disease found in Buddleja lindleyana Fort.usually was triterpene compounds. This experiment not only further arguing that buddleia fruit has certain research value in the research of treating Parkinson's disease from the Angle of molecular biology. |
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