Involvement Of Salvianolic Acid B To Blood Brain-barrier With Oxygen/Glucose Deprivation And Reintroduction In Vitro

Objective:In the past, more and more evidences indicated salvianolic acid B can protect the blood brain barrier against ischemia-reperfusion injury. However, there are many mechanisms in the process, and which involve the immune cells, inflammatory factors. How does the function of endothelial cells influence the process remain unknown. So we have established the BBB model in vitro, through lack oxygen and sugar, before regain sugar and oxygen treatment, immunofluorescence, electron microscope technology are used to observe the change. In the end, we discussed that salvianolic acid B influence blood brain barrier may be closely connected to the suppression of VEGFR overexpression.Methods:1. Used the brain microvascular endothelial cells and C6 cells non-contact co-culture to build blood-brain barrier model in vitro; We randomly divided the model into blank group, normal group, experimental group and drug group.2. We used sugar-free Earle ’s liquid instead of complete medium, and incubated the model in 93% N2,2%O2,2%CO2 environment for 2h to simulate the condition of lack of oxygen and sugar.3. Horseradish peroxidase permeability test to detect the changes of blood brain barrier (BBB) permeability.4. Immunofluorescence and electron microscope observation the changes in tight junctions between the endothelial.5. Reverse transcription polymerase chain reaction detected the changes in VEGFR-2 expression.Result:1. We have successful established the blood-brain barrier model in vitro, and the 4 hours test leakage test was positive2. The drug group’s HRP permeability is significantly lower than the experimental group, P< 0.05.3. The pictures of immunofluorescence and ultrastructure showed the drug group’s TJs were continuous and dense, the experimental group’s TJs were interruption and osteoporosis.4. RT-PCR results showed VEGFR-2 expression situation in each tine point: Experimental group> drug group> normal group, and the discrepancy was the most obvious on the third day, P< 0.05.Conclutions:1. Salvianolic acid B can protect the BBB directly from ischemia-reperfusion injury.2. Salvianolic acid B regulate the permeability of blood brain barrier through affecting the expression of VEGFR-2 and its signaling pathways downstream.