Nerve Protection Substance And Its Mechanism Study Of Shexiang Baoxin Pill

Shexiang Baoxin Pill (SBP), a composite formula of traditional Chinese medicine, consists of seven materia medicas, such as Moschus, Radix Ginseng, Calculus Bovis, Cortex Cinnamomi, Styrax, Venenum Bufonis and Borneolum Syntheticum. SBP is used for treating angina pectoris, chest distress and myocardial infarction due to myocardial ischemia, and widely used in clinical treatment for coronary heart disease. In recent years, many reports indicated that SBP showed a cignificent effect on cerebral ischemic diseases, with improving microcirculation, reducing cerebral vascular resistance, increasing cerebral blood flow, improving cerebral ischemia induced brain injury, diminishing the volume of cerebral infarction, and improving the symptoms of nerve injury. The active ingredients of SBP and the mechanism to protect the nerve injury were studied in this research.On the basis of early serum pharmacochemistry research, this study is designed to screen the serum compounds of SBP against glutamate induced PC 12 cells injury at the cellular level. The research demonstrated that gamabufatalin, muscone and cinnamaldehyde of twenty-six kinds of serum compounds could protect glutamate induced PC 12 cells injury, among which cinnamaldehyde was the most effective. Then, a further study focused on the in vitro effectiveness and working mechanism of cinnamaldehyde against glutamate induced PC 12 cells injury was carried out.Firstly, the effectiveness of cinnamaldehyde against glutamate induced PC 12 cells injury was evaluated by MTT method, flow cytometry, Hoechst 33258 staining and enzyme activity detection. The results showed that pretreatment with cinnamaldehyde at 5,10 and 20 μM could significantly attenuate cell viability loss, reduce apoptosis with dose-effect relationship.Secondly, the SOD activity, MDA level, GSH level and ROS content were also tested to evaluate the protective effect of cinnamaldehyde against glutamate induced PC 12 cells from oxidative stress injury. The experimental results showed that all concentrations of cinnamaldehyde could reduce the GSH, MDA levels and SOD activity, decress the generation of ROS in PC 12 cell.Lastly, pretreatment with cinnamaldehyde at 5,10 and 20 μM significantly stabilised mitochondrial membrane potential, decreased the release of cytochrome c from the mitochondria into the cytosol and limited the activities of the caspase cascades, such as caspase-9 and -3, also markedly increased Bcl-2 expression and decreased Bax expression, which demonstrated that the protective effect of cinnamaldehyde on glutamate-induced PC 12 cells injury was via mitochondria-mediated pathway.Taken together, these results indicate that cinnamaldehyde has potential protective effects against glutamate-induced oxidative stress in PC12 cells via mitochondria mediated pathway, suggesting that cinnamaldehyde might be one of the active ingredients in SBP for treatment of nerve injury induced by cerebral ischemic diseases, which also provide the scientific basis for SBP in the development of new clinical application.