Ginkgolide B The Molecular Mechanism Of Anti-platelet Aggregation In Aging Research

This study consists of two parts: literature summarization and experimental study, which will set a stong foundation for further clinical application.Literature summarization1. Literature research of Ginkgo biloba leaf Ginkgo biloba is considered as"living fossil"which is gymnosperm the only living species of the family Ginkoaccae.The dried leaf of Ginkgo biloba used as herb was first recorded in Bencao Pinhui Jingyao which said:"It belongs to the lung channel and can astringe the lungs to treat cough and asthma. It can also tonify the heart and relieve the pain".By reviewing literature from the past dynasties, we summarized Ginkgo biloba leaf's flavor, channel entered, distribution and effect which offer the theoretical basis for the research.2. The chemistry and pharmacology research of Extract of Ginkgo leafThere are about 180 kinds of chemical compounds in the Ginkgo leaf ,among which the two active ingredient of Extract of Ginkgo leaf are 24% flavonoid glycosides and 6% terpenes. Flavonoid mainly removes free radicals and anti-lipid peroxidation. Meanwhile, Ginkgolide B is the most effective receptor antagonist of PAF and can suppress the platelet aggregation and prevent coagulation. Ginkgolide B is the most effective among all terpenes and is widely used in clinic.3. Research about Thrombosis We try to explain the mechanism and onset of Thrombosis in molecular biology level and to find the key point of signal transmition pathway, which will help us to select experimental index.4.Research about Thrombosis by the influence of Aging With aging, the incidence of cardio-cerebrol vascular disease increased significantly and the cardio-cerebrol vascular systerm have taken place in some physiological changes. These changes often contribute to the formation of thrombus, however, the mechanism of age-induced changes in physiological functions is not very clear. We begin to study the molecular mechanism of thrombosis with aging, which is the hot issues.5.Traditional Chinese Medicine research regarding Aging and Thrombosis To find the relationship between Aging and Thrombus in TCM, the syndrome, pathogenesis and therapeutic principle will be treated based on differentiation of syndrome meanwhile the recent medication used for treating thrombus will be summarized.6.Research about Ginkgolide B's effect on anti-thrombus Summarize the recent experiment method about thrombus treatment by Ginkgolide B and the mechanism of Ginkgolide B's function. Experimental studiesThe inhibitory effect of Ginkgolide B on aging platelet aggregation Method: NO was measured by using a Nitric Oxide measurement kit.The phosphorylation of VASP was detected by Western blot. Platelet aggregation was measured by Chrono-log Platelet Aggregometer. ROS was measured by the flow of cytometry. The expression of NOX4 was determined by Western blot.Results: A high dose of Ginkgolide B(800μg/ml)obviously enhance the production of NO,but the small dose of Ginkgolide B(200μg/ml)was almost the same with the control. The expression of the p-VASP was decrease by Collagen, however, Ginkgolide B can inhibit the effect of Collagen and enhance the expression of the p-VASP. Ginkgolide B had obviously inhibitory effect of platelet aggregation induced by ADP,Collagen and H2O2 and can last the aggregating time which according to the concentration of dose and manner, but there was no inhibitory effect of aggregation induced by AA. Ginkgolide B inhibited the production of ROS induced by 0.01% H2O2 which increase the production of ROS. Meanwhile, Ginkgolide B (800μg/ml)inhibited the expression of phosphorylation of NOX4 induced by 0.01% H2O2.