Experimental Study Of Antiparkinsonism Activities Of Essential Oil Extracted From Alpinia Oxyphylla Miq.Fruit

Parkinson's Disease(PD) is a kind of gerontism nervous system degenerative disease.PD attack rate goes up year by year, which is second only to Alzheimer disease, and its disease incidence has already reached 1.03%.At present,there are few clinical drugs for PD treatment and there are shortcomings,such as curative effect decline and serious adverse effects after long-term administration,it is thus evident that it is important to development high efficiency low toxical drug for PD's prevention and cure.Alpinia Oxyphylla Miq.fruit is mature and dry fruit, which is one of the most famous traditional medicine in South-China.To speed up the development of Alpinia Oxyphylla Miq.,Chinese pharmaceutical researchers have undertook systematic researches on AOF.Recent study results indicated that the main chemical compositions in Alpinia Oxyphylla Miq. fruit were essential oil, microelement, terpene flavonoids, heptanes, etc,and essential oil accounted for 1%～2%. As a result, the keypoint to research the chemical composition is to research essential oil.More than 100 kinds of chemical compositions has been analyzed by Gas chromatography mass spectrometry(GC-MS) method.The main compositions were p-cymene, neral,coriandrol, myrtenal,a-pinene,β-pinene, etc,and the main microele- ments were Zn,Cu,Fe,Mn,Gu,Mg,Ca etc.Zn and Mn were obviously higher than other tonic.Modern medicine study has shown that Alpinia Oxyphylla Miq.fruit have the pharmacologic action on strengthen heart power, anti- cancer,anti-anapyhlaxis, anti-aging, cons- tious-sedation and relief pain. Recent years, more and more Alpinia Oxyphylla Miq.fruit as primary composition in some compound preparation has been applicated on clinical researches for intelligence and brain.It has been proved that Alpinia Oxyphylla Miq.fruit has good effects on brain protection in animal experiment, and can be used to treat senile dementia and other disturbance of intelligence. It also has antioxygen, anti-ageing, anti-apoptosis effects and can lessen excitability toxicity effect on nerve cell.As far as it is concerned that Alpinia Oxyphylla Miq. fruit is mainly produced in Hainan Island, where Alpinia Oxyphylla Miq.has been cultivated in large areas with high output and fine quality, it is feasible to extract essential oil from AOF for PD treatment.Objective:To study the effects and mechanisms of AOF on the experimental PD.Methods:(1)Wet distillation method was used to extract Essential oil from AOF;vapor-phase chromatography was applied to analyse the p-Cymene of essential oil extracted from AOF for studying the stability and normallability of essential oil extracted from AOF; (2)BLISS method was used to study acute toxicity test in mice and determine the LD50 ;(3) The C57BL mice were formed with intraperitoneal injections of 1-methyl- 4-phenyl-1,2,3,6-tetrahydropyri-dine(MPTP) to establish PD model; behavioral exhibition was explored by swimming test,water maze experiment and antonomic activities counting experiment;(4)Mice were formed with intraperitoneal injections of MPTP to establish PD model; the dopamine(DA), 5-hydroxytryptamine(5-HT) were abstracted by solvent extraction method; homovanillic acid(HVA) was abstracted by chroma- tographic analysis antigenic method;the DA,5-HT,HVA levels of striata were analysed by Fluorospectraphotometry method;observe essential oil extracted from AOF effect on monoamine neurotransmitter in mice brain; (5)The malondialdehyde(MDA),reduced glutathione hormone(GSH) and superoxide dismutase (SOD) activity from mice striatum were measured by ultraviolet spectrophotometer method to explore the mechanism of protect- ive effects;(6)The tyrosine hydroxylase(TH),B-cell leukemia- lymphoma- 2(BCL-2)and CASPASE-3 expressed in SNc were detected by Immunohisto- chemical SABC method;TUNEL staining method was used to detecte apoptosis in SNc of mice.Results:(1)The chemical compositions of essential oil extracted from AOF were stable and normallable; (2)The LD50 of essential oil extracted from AOF was 8.3269 ml·kg-1 and its 95% confident limit was 7.037-10.06 ml·kg-1,which suggests the toxicity of essential oil extracted from AOF might be minor;(3)Ethology experimental results showed that the AOF groups had significant difference with the MPTP model group;essential oil extracted from AOF could prolong swimming time of PD mice,shorten latency phase of mice in the water maze and decrease frequency of mice into blind-ending , but hadn't significant effect on autonomic activities of PD mice;(4)Essential oil extracted from AOF obviously antagonized MPTP-induced elevation of MDA,as well as decreased of GSH level and SOD activity of brain tissue; (5)Essential oil extracted from AOF obviously antagonize MPTP-induced decrease of DA,HVA,5-HT of PD mice; (6)Essential oil extracted from AOF could significantly increase the TH,BCL-2 expression,obviously decrease CASPASE-3expression in SNc, meanwhile inhibit nerve cell Apoptosis in SNc.Conclusion:(1)The compositions of essential oil extracted from AOF are stable and normallable;(2)The toxicity of essential oil extracted from AOF is very low;essential oil extracted from AOF could antagonize ethology change of PD mice induced by MPTP,which suggests that AOF can have effects on prevention and cure PD induced by MPTP.(3)It is showed that the anti-Parkinson's disease mechanisms of essential oil extracted from AOF might relate to the following factors. ①Essential oil extracted from AOF inhibits generation of oxygen- derived free radicals,and accelerates cleaning of free radicals,and relieve lipid peroxidation reaction, and maintains contents of SOD, GSH in normal level.②Essential oil extracted from AOF inhibits activity of monoamine oxidase-B(MAO-B), repress DA, 5-HT destructive metabolism.③Essential oil extracted from AOF could increase self-antioxygen capability, regulate the expression of modulin that intimate related to apoptosis such as BCL-2,CASEPASE-3,TH,etc.,then inhibits apoptosis.