Prevention Mechanism Of NaoJian Tablet On Cerebral Ischemic Injury And Its Pharmacokinetics

Based on the theories of modern medicine and traditional Chinese medicine, the purpose of this study was to investigate the prevention and its mechanism of NaoJian tablet during focal ischemic brain injury condition in vivo/in vitro.In vitro studies, MTT, ELISA, FCM and IF methods were used to determine the prevention effect of NaoJian Tablet and Buyang Huanwu Decoction by means of focal ischemic brain injury cell model, induced by treating PC12cell with0.3mM H2O2for2h. The result suggested that both of the two tested subjects could effectively protect PC12cell from impairment induced by H2O2in the range of0.2～1.0mg/mL. Moreover, we found not only the reduction of L-LDH in cell culture medium but also inhibition of the basal cellular ROS and CDK5/Tau levels following exposure to NaoJian Tablet from Buyang Huanwu Decoction, Conversely, the increasing cellular GSH content been detected. The all results reveal the reasonable of selection of NaoJian Tablet. Moreover, the decomposing experiment of NaoJian Tablet show that Astragalus membranaceus (Fisch.) Bunge, Rhizoma Chuanxiong, and Pheretima aspergillum (E.Perrier) in the ratio of10:3:1is consistent with the principle of science of traditional Chinese formula. The concentration main bioactive suhbstance such as ligustrazine, ferulic acid, senkyunolide and ligustilide in NaoJian Tablet was determined simultaneously by use of HPLC-DAD method for its overall quality evaluation.In vivo studies, immunohistochemistry, RT-PCR and Western blot were applied to determine the expression of CDK5、CDK4/CyclinD1in the hippocampus tissues in MCAO rat by treating with NaoJian Tablet and Buyang Huanwu Decoction. The result indicated that NaoJian Tablet (ig,2.41g/kg) could significantly decrease the levels of CDK5、CDK4/CyclinD1in hippocampus tissues in MACO rat after treated by3d,7d,14d,28d (p<0.05). The similar mechanism was observed in the Buyang Huanwu Decoction treated MACO rat (p>0.05). The above-mentioned in vivo studies suggested that NaoJian Tablet maybe play its prevention effect on nerve cells via its acting on CDK5signaling and cell cycle pathway, which verified the multi-target and multi active pathways efficiency of tradition Chinese medicine in treating diverse disease. On the other hand, the processes of metabolism and disposition of several active components in NaoJian Tablet and Buyang Huanwu Decoction were analyzed and characterized using HPLC-DAD methods. The data show that all of the three tested compounds were fitted to two-compartment metabolic model. The three tested compounds in NaoJian Tablet maintained its own metabolism regularity owing to the discrepancy in pharmacokinetic parameters with Buyang Huanwu Decoction. The present study mainly focus on the preclinical research of NaoJian Tablet. The results lay a foundation on the registration application of it. Simultaneously, it can conduct as a reference of research approach for traditional Chinese medicine.