The Research On Borneol Opening Blood Tumor Barrier Molecular Of Mechanism

Objective:Establish a blood tumor barrier (BTB) model in vitro to study the molecular mechanism of opening blood-brain barrier by medicine borneolMethods:1.Isolation, cultivation, identification of human umbilical vein endothelial cells.2.Establish blood tumor barrier model in vitro by non-contact co- cultureing of C6 cells and the third generation HUVECs.3.Observed respectively the changes of expression of claudin-5 of HUVECs,a non-contact co-culturing with C6 cells and the HUVECs as blank control culture by using Immunohistochemistry. To observe 3rd generation cultured alone the tight junction formation of HUVECs by Projection electron microscope.4.Applied20μg/mlofBorneoltothemodelof blood tumor barrier,after 2h,4h,8h,16h,24h,the expression of claudin-5 of the model of blood tumor barrier was detected by immunohistochemistry on HUVECs. Detect tight junction of the model of the blood tumor barrier in vitro by electron microscope in the same time.Results:1.FactorⅦin HUVECs showed positive by Immunohistochemistry,successfully to Isolation, cultivation, identification of human umbilical vein endothelial cells.2. Successfully to establish blood tumor barrier model in vitro.3. Detection of the blood tumor barrier in vitro:3.1 Immunohistochemistry showed:non-contact co-culture in vitro model of blood tumor barrier tight junction protein claudin-5 expression, compared with HUVECs cultured alone on the expression of claudin-5 was significantly increased (P<0.05). 3.2Transmission electron microscopy showed:non-contact co-culture in vitro model of blood tumor barrier tight junctions, HUVECs cultured alone more closely on the continuous and close connection.4.The results of Acting on the the blood tumor barrier model in vitro by borneol:4.1Immunohistochemistry showed:the claudin-5 expression was not changed significantly in each group of blood tumor barrier in vitro after the concentration of 200ug/ml of borneol. (P>0.05).4.2Transmission electron microscopy showed:After 4 hours, the blood tumor barrier tight junction on the model began to be destroyed by borneol, to 16 hours the most serious destruction was observed,after 24 hours continuous tight junction recovery and become more closely.conclusion:1.Successfully to establish blood tumor barrier model in vitro2.Borneol can open the blood tumor barrier by influencing the formation of tight junctions.in this study, the changes in tight junction barrier,was not found after 4hours the role of borneol in the first blood tumor barrier, but after 16 hours the start was found that the blood tumor barrier tight junction becomes sparse and not continuous. After 24 hours of continuous tight junction recovery has become more closely. 3.Borneol opening blood tumor barrier mechanism is not interfere on the expression of claudin-5 of HUVECs to achieve.