| Foshousan, also called xiongqiong decoction, is composed of Angelica sinensis and Ligusticum chuanxiong, with well formula and strict compatibility. Which has the pharmacological action of taking account both Qi and blood and activating blood circulation and Qufeng and relieving pain. Which were used to treat irregular menses, postpartum blood deficiency, dizziness, headache and some aches in heart and stomach in ancient times. Modern pharmacology has confirmed that the prescription can inhibit hyperplasica of vascular smooth muscle, improve blood circulation, inhibit platelet aggregation and anti cerebral ischemia and has antioxidant effect etc. Which has been widely used in treatment of cerebrovascular disease and cardiac disease in recent years with unique advantages. In spite of foshousan and it’s single herb and their active ingredients have been widely used in clinical. Effective material foundation of foshousan decoction prescription for treating cerebral ischemia injury has not been reported systematically. We have fully taken advantage of intersecting of various subjects and had research on the chemical composition of foshousan decoction for therapy cerebral ischemia injury with modern means of science and technology.Firstly, the foshousan decoction was prepared with optimized extraction technology. Secondly, taking the10batches foshousan decoction samples as object, foshousan decoction fingerprints was established by HPLC method. Fingerprints’ mutual model and similarity among10batches were established with the software of "fingerprint similarity evaluation system in China (2004A edition)". The results showed that common peaks’relative retention time of each batch of samples were almost the same, RSD is less than1%, similarity were greater than0.94, Which are accorded with the technology requirements of Chinese traditional medicine fingerprint. And the chemical composition of foshousan decoction was characterized with correlativity analysis between the HPLC fingerprints of single herb and that of combination decoction.The chemical composition of foshousan decoction was analyzed by UPLC-ESI-MS technology furthermore based on the earlier work.11components were successfully identified by the analysis of data from MS spectrum and UV spectrum based on comparing with several standard compounds and references. Which were chlorogenic acid, caffeic acid, ferulic acid, chuanxiongol, senkyunolide I, senkyunolide H, butylidenephthalide, butyl phthalide, senkyunolide, Z-ligustilide and dibutyl phthalate. Main chemical material foundation has been identified preliminary.The established mutual model can well reflect the distribution of full quality and quantity for the chemical composition, which can provide a reference basis for the medical’s efficacy material base research and it’s quality control. Literature shows that ferulic acid, senkyunolide I, senkyunolide A and Z-ligustilide have been proved to be the active components for cardiovascular and cerebrovascular function. So the methods used to determine the four active compounds simultaneously by UPLC was established on this research. The results shows that the method is simple, high precision, which can offer a scientific basis for controlling quality of the foshousan decoction.In order to further confirm the curative effect on prevention and treatment for ischemic stroke of foshousan decoction, this study combining different levels of cerebral ischemia injury in vivo and in vitro models, respectively research the foshousan decoction’s protection for ischemic brain injury from two levels of the whole animal and cell.The middle cerebral artery occlusion(MCAO) model in rats was established by line plug method. The protection effect on experimental cerebral ischemia injury was observed by the whole animal’s level. The results showed that foshousan decoction has a significant function on improving neurobehavioral disorder caused by experimental cerebral ischemia, degree of brain edema and the volume of cerebral infarction, and the effect was dose dependent, which proves that foshousan decoction has good protection on cerebral ischemia-reperfusion injury. It’s mechanism can be connected with increasing the antioxidant enzyme activity in brain tissue and reducing lipid peroxidation.In addition, the model of oxygen-glucose deprivations (OGD) in vitro PC12was established with combining oxygen-deprived by sodium dithionite and low glucose. The protection on PC12nerve cells with oxygen-glucose deprivation injury was observed from the cellular level. Results show that compared with model group, the foshousan decoction pretreatment group has obvious protective effect on the cell damage by oxygen-glucose deprivation, and which can reduce MDA and NO levels with dose dependent and increase SOD activity of cells. It maybe one of neural protection mechanisms by foshousan decoction.On the basis of experiments in vivo and in vitro. Three extraction parts including ethyl acetate, n-butyl alcohol and residual water extraction part were obtained by using different solvents to gradient extraction of foshousan decoction. The comparative research of foshousan decoction and different polar extraction parts on anti cerebral ischemia was carried out by OGD model. The results show that the extraction parts of ethyl acetate and residual water have certain protective effect on nerve cell damage, which maybe the effective parts of anti brain ischemia. Based on the analysis of chemical composition in the earlier research, it showed that ferulic acid, senkyunolide I, senkyunolide A and Z-ligustilide maybe the main effective components of foshousan decoction against cerebral ischemia. Due to lack of reference standards and the experimental conditions restrictions, some water-soluble ingredients in prescription were failed to identify, which needs to be further study.This study can provide a scientific basis for explaining the effect on anti cerebral ischemia, the chemical composition and establishing quality evaluation method of foshousan decoction. Which can provide a theoretical reference for this prescription’s secondary development and clinical application. |
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