The Pharmacodynamic Substance Of Oils Of Angelica Sinensis And Ligusticum Chuanxiong For Protecting Brain Injury

Objective:Based on traditional Chinese medicine chemical methods and pharmacological activity screening means,chemical composition of oils of Angelica sinensis and Ligusticum chuanxiong were separated and identified,and to explore protective effects of oils of Angelica sinensis and Ligusticum Chuanxiong for protecting brain injury,and to look for efficacy material basis.Methods:1.By water vapor extraction,oils of Angelica sinensis and Ligusticum chuanxiong were extracted,then by GC-MS method,composition of oils of Angelica and Chuanxiong were detected.2.Using various chromatographic packings,chromatographic methods and spectroscopic methods for isolation and identification of chemical constituents of the two oils.3.NaNO₂ was used to set up acute chemical brain injury,and the brain tissue was taken out to detect levels of Na⁺-K⁺-ATP,Ca²⁺-Mg²⁺-ATP and AchE.Besides,MTT method was used to measuring effects of the two oils on PC12 cells,and cell survival rates of compounds on PC12 cells injuied by glutamate and CoCl₂ model were detected.And then lactate dehydrogenase?LDH?was used to test levels LDH in PC12 cells.Results:1.There were 14 compounds and 17 compounds determined in oils of Angelica sinensis and Ligusticum chuanxiong.The main compositions include phthalides,sesquiterpenes and aliphatic compounds.For the content of main compositions,content of ligustilide was the highest?54.36%?,Z-butenylphthalide was16.81%in oil of Angelica sinensis,ligustilide was 63.88%and terpinen-4-ol was 9.79%in oil of Ligusticum chuanxiong.2.By systematic chemical separation,15 compounds were identified from oil of Angelica sinensis,the main type of those compounds is butylphthalides,including two newcompounds,?+?-?R?-3-butyl-7-hydroxy-3-methoxyphthalide?1^*?and Senkyunolide I-6,7-acetonide?2^*?.And from oils of Ligusticum chuanxiong,7compounds were found which contained 4 butylphthalides,one sesquiterpene and two aromatic compounds.3.Oils of Angelica sinensis and Ligusticum chuanxiong can improve levels of Na⁺-K⁺-ATP and Ca²⁺-Mg²⁺-ATP,and reduce levels of AchE in mice'brains injured by NaNO₂,and no inhibition was found to PC12 cell viability.At 6.25?g/mL and12.5?g/m L,oil of Angelica sinensis can improve cell viability injured by glutamic acid?P<0.01 or P<0.05?.At 3.125,6.25 and 12.5?g/m L,the protecting effect of oil of Ligusticum chuanxiong was incremental trend?P<0.01?.For compounds 1^*,4,5 and16,at a certain concentration,can improve survival rate of injured PC12 cells?P<0.01or P<0.05?.For PC12 cells injured by CoCl₂,at 3.125,6.25,12.5 and 25?g/ml,oil of Angelica sinensis can improve cell viability?P<0.01?,at 3.125,6.25,12.5 and 25?g/ml,oil of Ligusticum chuanxiong can improve cell viability?P<0.01?.For compounds 3,7,10,11 and 16,can improve cell viability at a certain concentration?P<0.01 or P<0.05?.Besides,oils of Angelica sinensis and Ligusticum chuanxiong and compounds 16 and 11 can reduce level of LDH?P<0.01?.And after dyeing of Hochest,compared with model group,at concentration of administration of oils of Angelica sinensis and Ligusticum chuanxiong,and compounds 16 and 11,the cell morphology was more complete,and cell fluorescence intensity was weaker.Conclusion:1.The principal component type between oil of Angelica sinensis and oil of Ligusticum chuanxiong were similar.And the difference was on trace elements.2.After component separation and structural confirmation,phthalates from oils of Angelica sinensis and Ligusticum chuanxiong were similar,and we should note instability of oils during the component separation.3.Oils of Angelica sinensis and Ligusticum chuanxiong can regulate levels of ATPase and reduce level of AchE,and were the important section for protecting brain injury.And compounds 1^*,3,4,5,7,11 and 16 can protect injured PC12 cells by reducing level of LDH and inhibiting cell apoptosis.