| Purpose:The senile dementia, also called Alzheimer's disease or AD for short, is a kind of degenerative diseases of nervous system which occurs to the old age crowd's .The patient displays for the carrying on memory obstructs bonds, the spirit and the behavior exceptionally, lose the ability of take care of oneself , creates the tremendous pressure for the family and the society. As progressive along with the society, the global population old age degree inceasingly enhances, but AD more and more becomes the old age society senior citizen life and the quality of life huge threat. It has become a harm to life in European and American developed country AD the fourth big disease;In China, above 60 years old the population surpasses 100million, the statistics demonstrated that, AD above 60 years old in population prevalence rate approximately 5.0%, moreover grows the remarkable rise along with the age, above 80 years old the population prevalence rate may reach above 20%. Therefore, the related research on AD in domestic and foreign has been hot spots topic, which has the significant social significance.After the scientific research personnel's joint effort, had the preliminary understanding to the AD pathology pathogenesis, and has formed many theories hypotheses,Mainly includes: 1, amyloid beta-protein theory. 2, the choline can damage theory. 3, the free radical damage theory. 4. calcium balances are out of balance theory. 5, gene mutation theory. 6, endocrine disorders theory. 7, inflammation response theory.The existing theory hypothesis although has its reasonable one side, but has the certain limitation. There is none of them to be allowed completely to elaborate the AD pathology mechanism. The massive research demonstratedthat, the AD itself formation reason is extremely complex. At present still lacked the effective method regarding the AD medicine treatment, used the treatment AD plan according to the above hypothesis, the clinical curative effect was not certainly ideal. To threatens facing AD, our country scientific research workers also conduct the widespread research to the Chinese medicine preventing and controlling senile dementia, has done the massive work. The clinical research and the animal experimentation result showed the Chinese medicine to prevents and controls the AD very good prospect.This topic group Yangshoudan to the Chinese native medicine tradition compound prescription to carry on the massive experimental study work, already completed the research included: To in the body animal, the application many kinds of methods creates the AD pathology experimental model, from the behavior study, the morphology, the biochemistry target examination, handed over aspect and so on nature acceptor change has conducted the massive research. The result indicated that, raises Yangshoudan to be allowed obviously to improve model animal behavior study unusual, sharpens the study memory ability, overcomes the experimental spatial cognition barrier. Enhances in the brain the memory dependence nerve to hand over the nature content level, in experimental animal brain organization shape unusual which the improvement pathology factor creates, strengthens the brain to organize the anti- oxidation to be supposed to stir up with free radical harm ability and so on.This topic in raisesYangshoudan to prevent and control the AD series research in the foundation, uses the blood serum Pharmacology method, in view of AD in the neuron massive losses pathology change, manufactures the cell model by amyloid beta-proteinwhich the PC12 cell weakly perishes, the application modern cytology, the molecular biology technical method, from the cell member, the gene protein level to raisesYangshoudan to prevent and control AD the function and the action mechanism carries on the discussion.Method: (1) raisesYangshoudan to withdraw the thing according to the traditional process method preparation, the tradition for the medicine way and the method, namely fills the stomach for the big mouse to raiseYangshoudan to withdraw thing every day two, continuously three days, after fourth days first time for a medicine hour, take the big mouse blood preparation blood serum to use in the blood serum pharmaceutical chemistry analysis and thecell raise research. (2) Yangshoudan blood serum at the beginning of pharmaceutical chemistry to search: Uses the highly effective liquid chromatography (HPLC) , Yangshoudan to contain the medicine blood serum, the blank blood serum and raises theYangshoudan withdrawing chemical composition to carry on the comparative analysis, to the time discovered contains the medicine blood serum drug efficacy ingredient and its the origin, promulgates in the Chinese native medicine compound prescription in vivo process the chemical composition change and the medicine effective component. (3) amyloid beta-proteinsends the PC12 cell apoptosis and to raise theYangshoudan' s protective function: Using in vitro cell raise technology, fosters together A and the PC12 cell, may obtain the similar AD pathological change the nerve cell apoptosis the model, Weakly perishes in the preparation cell in the model, joins raisesYangshoudan to contain the medicine blood serum to carry on the raise, this research chose the electricity mirror to observe the PC12 cell the ultramicro shape, the MTT law and the class tries the cell technology examination cell apoptosis the degree. This research chose the electricity mirror to observe the PC12 cell the ultramicro shape, the MTT law and the class tries the cell technology examination cell apoptosis the degree. Observes the occurrence situation as well as the medicine from the cellular form and the biochemistry change which the PC12 cell weakly perishes processes the influence which weakly perishes to the cell. (4) Yangshoudan send the protective function mechanism research to amyloid beta-proteinwhich the PC12 cell weakly perishes. Foster together in the PC12 cell and A in the cell model, simultaneously joins with A raisesYangshoudan to contain the medicine blood serum, with flow cytometry to examine itrocellar Ca content in the PC12 cell;In immunity group method determination cell or nutrient fluid NO, NOS;Uses the enzyme association immunity adsorption (the ELISA ) to examine Caspase-3, the Bcl-2 content;the NF-kB expression was detected with an epifluorescence microscope, from weakly perishes by the time the gene regulation level to promulgate Yangshoudan apoptosis to the nerve cell the protective function mechanism.Result: (1) Yangshoudan to contain the medicine blood serum pharmaceutical chemistry research demonstration, the Chinese native medicine after in^rivo process, the chemical composition has the quite big change. The newingredient in the blood serum appears which under this experimental condition has not seen from the raw material for medicine primary form ingredient, but in vitro alone withdraws the raw material for medicine its chemical composition actually can withdraw in the thing in the compound prescription to appear, And by Bajitian, Roucongrong and polygonum multiflorum's ingredient originates many, may explain the Chinese native medicine compound prescription theory rationality as if in the certain degree. (2) Ap and the PC12 cell raise, appeared the shape change which the model cell weakly perishes, If the model group PC12 cell obviously cell membrane periphery filament wool vanishes, the cell shape changes the circle, cellar jam in appears the vacuole, But Yangshoudan to contain the medicine blood serum group then the shape change is not obvious. The MTT law and flows the type cell meter to examine the PC12 cell apoptosis has the situation, demonstrated the medicine processing group cell weakly perishes compared to the model group reduces, compared to the has not been remarkable with the normal group difference. (3)In the cell the Ca2+ examination result, model group A created in the PC12 introcellar Ca2+ overload, the medicine processing group has been allowed to resist weakly perishes in the cell the Ca2+ serious to be unbalanced;And the NOS examination discovered to PC12 cell NO that, the model group NO content and the normal group a are more remarkable than increase, the NOS expression also similar enhancement, raisesYangshoudan and so on various medicines processing group two targets both to have the varying degree the improvement, causes NO, NOS to produce with the expression all drops. Model group' s PC12 cell Caspase-3 activeness and the comparison group a are more remarkable than enhancement, raises medicine processing group Caspase-3 activeness and so on Yangshoudan then the varying degree reduces;Model group nuclear factor -kB, the Bcl-2 determination result quite normal comparison group remarkably reduces, is raising medicine and so on Yangshoudan with the processing experimental group, nuclear factor -kB, the Bcl-2 activeness is all enhanced.Conclusion: Yangshoudan to contain in the medicine blood serum to compare the blank blood serum to increase the chemical composition, at under this experimental condition, contained in the medicine blood serum the discovery not to originate to the compound prescription Chinese native medicine withdraws in the thing the primary form ingredient, prompted the Chinesenative medicine in the process in vivo process the ingredient change to be bigger, The Chinese native medicine medicine generation of dynamics research also needs to do the massive basic research, the research Chinese native medicine function and the action mechanism, seeks the drug efficacy ingredient also to be supposed to strengthen with the blood serum pharmaceutical chemistry research union. amyloid beta-proteinmay create the PC12 cell weakly perishes, raisesYangshoudan to contain the medicine blood serum apoptosis to the experimental PC cell has the certain protective function, displays for may suppress the cellular form which amyloid beta-protein causes apoptosis pathology change;May enhance apoptosis under the factor function cell vigor, the occurrence which the suppression9+cell apoptosis;Yangshoudan to be allowed to improve the PC12 cell the Ca to be balanced, suppresses in the cell the calcium overload to cause the PC12 cell apoptosis;Yangshoudan to be allowed to reduce under amyloid beta-proteinfunction the cell NO massive productions and reduces the NOS activeness, prevents PC12 cell harm which NO creates;Yangshoudan also can decline Caspase-3 apoptosis has the before expression, prevents apoptosis the joint level effect which induces because of the Caspase-3 expression;This experiment also discovered that, Yangshoudan to apoptosis has the function which in process negative feedback regulatory gene Bcl-2 and nuclear factor -kB has surrenders to the state, Yangshoudan to contain the medicine blood serum protection nerve cell, suppresses weakly perishes occurs possibly is Yangshoudan to prevent and control AD the pharmacological action mechanism. |
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