Neuroprotective Effect Of Buyang Huanwu Decotion On PC12 Cell Injury Induced By PAF

Cerebrovascular disease is common and frequently-occurring disease, which is harmfull to human life and health. It is one of three major causes of death. Among survivors,50% to 70% of patients have paralysis, aphasia, and other serious disability, which bring heavy burden to society and family. Finding new treatment methods has become a hot field of neuroscience. Nerve cells are composed of the basic functional unit of the nervous system, the maintenance of normal metabolic activity is highly dependent on the sustained and stable blood supply. Any systemic or local factors can make cerebral blood flow decrease to a certain threshold and cause nerve cell damage. Neuroprotective therapy aims to improve the nerve cells of the internal environment caused by any reason, tolerance and immunity disorders, block the different aspects of nerve cell necrosis,and increase the viability of cell in the pathological enviroment and promote nerve cell function recovery. With the confirmarion of thrombolysis and anticoagulation for treatment of acute ischemic stroke, protection effect of nerve cells has more and more important clinical significance.Platelet activating factor (PAF), is a biologically active lipid transmitters, the process of platelet aggregation induced by adenosine diphosphate does not depend on (ADP) or arachidonic acid (AA) metabolism Product of TXA2. That platelet aggregation induced by PAF is considered to be the third way. PAF takes part in a variety of pathological and physiological processes. Interactions of vascular endothelial cells and platelets plays an important role in the thrombosis. Relation of PAF and ischemic cerebrovascular disease is getting more attention. From atherosclerosis to stroke recovery on all aspects of PAF may play an important role. In order to prevent ischemic cerebrovascular disease, inhibition of PAF-induced pathological process is necessary, which may be an important way to the application of PAF inhibitorsCerebrovascular diseases in modern medicine is called stroke by traditional Chinese medicine(TCM). TCM treatment of stroke has a long history.the overall concept of syndrome differentiation based on stroke prevention, treatment, rehabilitation has a unique concept, accumulated a large, rich theoretical knowledge and treatment experience.Buyang Huanwu Decotion(BHD) is to include in the "Yi Lin Gai Cuo" book by Wang Qing Ren(famous doctor of Qing Dynasty).BHD is made up by Astragalus, Angelica tail, red peony root, earthworm, Chuanxiong, peach kernel, safflower. BHD's composition and building are based on the vigour deficiency, vigour can not be handsome blood, leading to blood stasis. BHD have effect of invigorating vital energyinvigorating vital energy, activating blood circulation and dredging collaterals and widely used in clinical treatment or prevention of ischemic cerebrovascular disease, coronary heart disease, myocardial infarction and other thromboembolic disease,is-the most frequent prescription applications of the treatment of stroke and other thrombotic diseases. Experimental Study of 1999 found that BHD has a neuroprotective effect. BHD was mainly on anoxia apoptosis level of protection and can promote peripheral nerve regeneration, neural stem cells growth, survival, and to neurons and glial cell differentiation under hypoxic conditions for conventional and neural element showed a significant role in promoting cell growth and promote the growth of embryonic neural epithelial cells, but also to promote their differentiation into neurons and glial cells. Currently nerve cell injury has been cured by Chinese medicine and made some achievements for the prevention and treatment of nerve cell damage accumulated some theoretical basis. We also found that BHD could improve spinal cord injury (SCI) hindlimb motor function and reduce PAF concentration of endogenous rat SCI, spinal cord injury to play on the protective effect of plasma in the lower SCI PAF content in hindlimb motor function and improve related. Protective effect on spinal cord injury may be related to reduce the number of PAF-R and inhibit PAF-R activity, thereby blocking the effect of PAF on to play injury.Due to nerve cell culture studies in ischemic brain injury in the advantages, such as research object is a living nerve cells, of the conditions can be artificially controlled studies of nerve cells can be achieved relatively homogeneous sample to study the contents of the ease of observation, detection and records, and fully meet the requirements of the experiment and research expenses and other than the economy, thus using a cellular model of neuronal injury Protective effect of nerve cells is particularly important. According to ischemic neuronal injury in the pathophysiology of proven, we established model of hypoxic neuronal damage observed on PC 12 cell injury model of hypoxic cell morphology, viability, and PAF content of the supernatant. Of drug serum in BHD on PC12 cells in hypoxic injury in rats. To further study effect of nerve cell injury, reflected the party on the nerve cell injury in many aspects of intervention, selected on the nerve cell injury has a protective effect of active ingredients to provide experimental basis. Methods:1.experimental animals, grouping and model:general level 6 New Zealand rabbits were randomly divided into:normal group 2 (distilled water group), drug serum in BHD group 2 (BHD group), positive control group 2 (Ginaton group): animals in each group were fed the same volume 10ml/kg,1/d, a continuous 7 d. 2.specimen collection and research method:fast before the last administration,12 h, 2h after the administration of blood (the central ear artery) to prepare drug-containing serum. Integration will have 80% of the PC 12 cells which were divided into 5 groups BHD containing serum group (2.5%), positive control group (containing serum ginaton 5%), blank serum (5% normal rabbit serum), model group (without rabbit serum), normal group (normal training including cattle, horse serum 5% each), grouped by the corresponding medium containing serum replacement. Dosing will be grouped in the PC12 cells were 37℃,5% CO2 and 90% nitrogen of the three gas incubator, the production of hypoxic model.①PC 12 cell morphology, hypoxia 1.5h, 3h,18h, the cells were observed by inverted microscope, morphological changes, and shooting with the image acquisition system;②After 18 h hypoxia modeling, MTT cell viability assay;③missing After 1.5 h of oxygen modeling, Annexin V/PI apoptosis detection;④1.5 h after hypoxia modeling, enzyme-linked immunosorbent assay (Elisa) culture supernatant PAF content.Results:①The morphology of the hypoxic injury of PC12 cells:the major manifestations of acute hypoxic injury 1.5h, compared with the normal group, regardless of dosing or, in the morphological features, and are disordered cell arrangement, cell morphology variation, and more irregular part of the connection broken, and with some off the wall, the gap between the larger cells,BHD group close to the normal cells. Hypoxic cells 18 h showed the self-repair, compared with model group, the cell growth-intensive, dosing each group have different levels of repair, cell bodies extended synapses, short thicker than the normal group, to re-establish the connection between the cell body However, a little disorder than in normal group, but BHD group and cell morphology Ginaton group close to the normal group.②The hypoxic injury of PC 12 cell viability:drug serum in BHD group was significantly higher cell viability, and Ginaton had no significant differences, but the blank model group was obviously increased, and the difference statistical significance (P=0.000);③Hypoxic injury rate of apoptosis in PC 12 cells: Ginaton group, blank serum group, model group were significantly higher than the normal cells group, the difference was significance (P=0.000), but 2.5% BHD serum apoptosis rate compared with the normal group the difference was not statistically significant (P=0.209), and significantly lower than the model group, there are significant differences. (P=0.000);④PC 12 cells against hypoxic injury PAF content in the supernatant:BHD group Ginaton and supernatant PAF group were significantly lower than the model group, the difference was statistically significant (P=0.000).Conclusion:BHD containing serum has obvious protective effect on PC 12 cell by hypoxia. Its mechanism is related to reduce the release of the content of PAF in cell by hypoxia, protect the cell shape and enhance the viability.