| Objiect:By making use of modern analyzing and detecting technique and microdialysis technique, to study the effect/toxicity amphiprotic components-ephedra alkaloids of Ma-Xing-Shi-Gan Prescription (MXP) in vivo and change in vitro brought by traditional use in ancient times.Methods:(1) A HPLC-UV method was established for the simultaneous determination of ephedra alkaloids in Ephedrae herbal. Furthermore, by Independent-Samples T Test, contents of ephedra alkaloids in fresh Ephedrae herbal were compared to those in shopworn Ephedrae herbal, respectively.(2) A HPLC-UV method was established for the simultaneous determination of ephedra alkaloids in Ma-xing-shi-gan decoction and Ma-huang decoction. Furthermore, by Independent-Samples T Test, contents of ephedra alkaloids in Ma-xing-shi-gan decoction were compared to those in Ma-huang decoction, respectively.(3) A Liquid chromatography-electrospray tandem mass spectrometry method (HPLC-ESI-MS/MS) was established for simultaneous determination of amino acid neurotransmitters in specific brain regions of rats, meanwhile, another HPLC-ESI-MS/MS was established for simultaneous determination of ephedra alkaloids in specific brain regions of rats. Furthermore, the contents of amino acid neurotransmitters were compared between deferent brain regions by One-way ANOVA and repeated measures. Also the contents of ephedra alkaloids were compared between deferent brain regions by the same method of One-way ANOVA and repeated measures.(4) By the help of microdialysis and rat brain stereotaxic apparatus, blood dialysis and brain dialysis was collected simultaneously, so that the PK-PD model was set up. With ephedra alkaloids as pharmacokinetic index and amino acid neurotransmitters as pharmacodynamic index, the total and part pharmacokinetic and PK-PD combination were studied.Results:(1) Ephedra alkaloids as the effect/toxicity amphiprotic components, the contents in shopworn Ephedrae herbal was less than those in fresh Ephedrae herbal (P<0.05). The content of NME, NMP, E, PE and ME in shopworn Ephedrae herbal was 0.42, 1.00,7.51,3.66,1.41 mg/g respectively. The content of NME, NMP, E, PE and ME in fresh Ephedrae herbal was 0.62,1.40,10.43,5.72,1.98 mg/g, respectively.(2) Ephedra alkaloids as the effect/toxicity amphiprotic components, the contents in Ma-xing-shi-gan decoction was less than those in Ma-huang decoction (P<0.05). The concentration of NME, NMP, E, PE and ME in MXP was 69.30,110.84, 2806.39,1171.80,464.51μg·mL-1, respectively. The concentration of NME, NMP, E, PE and ME in Ma-huang decoction was 159.87,272.70,4328.35,1810.12,532.86μg·mL-1, respectively. (3) After administration of the same contents of Ephedra alkaloids in deferent drug from, Ephedra alkaloids in specific brain regions were lower in MXP group than that in control group. Homologuely, the level of glu, asp in MXP group than that in control group. Therefore, the dose-effect relationship exist between the effect/toxicity amphiprotic components-Ephedra alkaloids and erethisma amino acid neurotransmitters.(4) In vivo effect study,1.5h after administration of MXP or control medicine, ephedra alkaloids in frontal were more than those in hippocampus and corpus striatum. After administration of the same contents of ephedra alkaloids, contents of ephedra alkaloids in MXP group were less than those in contro group in each brain region.(5) After administration of the same contents of ephedra alkaloids, contents of ephedra alkaloids in MXP group were less than those in contro group in each brain region. More over, the levels of excited neurotransmitters (Glu and Asp) in MXP group were less than those in control group in each brain region. The dose-effect relationship existed between concentration of ephedra alkaloids in brain and the level of excited neurotransmitters (Glu and Asp).(6) PK-PD model was maturate and scientific.(7) In PK-PD study, ephedra alkaloids could be absorpted into blood and brain in 30min after administration. Among the three groups, ephedra alkaloids had the same absorption behavior in blood and brain, respectively.(8) Among the three group, ME eliminated more quickly than E and PE, which eliminated more quickly than NME and NMP.(9) after administration of MXP or control medicine, the levels of Glu and Tau increased obviously.(10) A HPLC-UV method for simultaneous determination of five ephedra alkaloids in Maxingshigan decoction was set up.(11) A HPLC-ESI-MS/MS method for simultaneous determination of five ephedra alkaloids in brain homogenate was set up.(12) A HPLC-ESI-MS/MS method for simultaneous determination of amino acid neurotransmitters in brain homogenate was set up.(13) A HPLC-ESI-MS/MS method for simultaneous determination of five ephedra alkaloids in blood and brain dialysis was set up.(14) A HPLC-ESI-MS/MS method for simultaneous determination of amino acid neurotransmitters in brain dialysis was set up.Conclusion:Chemical study in vitro, contents of ephedra alkaloids in shopworn ephedrae herbal were less than those in fresh ephedrae herbal, respectively. Meanwhile, the contents of ephedra alkaloids in MXP were less than those in Mahuang decoction. In vivo effect study, after administration of the same contents of ephedra alkaloids, contents of ephedra alkaloids in MXP group were less than those in contro group in each brain region. More over, the level of excited neurotransmitters (Glu and Asp) in MXP group were less than that in control group in each brain region. The dose-effect relationship existed between concentration of ephedra alkaloids in brain and the level of excited neurotransmitters (Glu and Asp). In PK-PD study, ephedra alkaloids could be absorpted into blood and brain in 30min after administration. Among the three groups, ephedra alkaloids had the same absorption behavior in blood and brain, respectively. |
PDF Full Text Request