Protective Effect And Mechanism Of Gastrodin Injection On HBMEC Damaged By OGD

Objective:To explore the protective effect and mechanism of gastrodin injection on HBMEC damaged by OGD.Methods:1.HBMECs were cultured in vitro to establish a stable cell experiment platform.2.HBMECs were randomly divided into 4 groups:normal group,model group,nimodipine injection groups and gastrodin injection groups.In addition of the nomal group,OGD was used in other groups.The nimodipine injection groups were treated with different concentrations(11.950,2.390 and 0.478 ?M)of nimodipine for 24h prior to OGD and the process of OGD.and the gastrodin injection groups were treated with different concentrations(3.386,0.677 and 0.135 mM)of gastrodin for 24h prior to OGD and the process of OGD.Cells ware treated with OGD for 1,3,6 and 9 h.The viability of HBMECs was measured by MTS.3.Claudin-5,Occludin and ZO-1 mRNA transcription and protein expression levels in HBMEC damaged by OGD were measured by qPCR and Western blot.Results:1.When the density of HBMEC reached 90%,the cells were damaged by OGD for 9 h.Then the cell viability measured by MTS is decreased to about 57%.2.After the density of HBMEC reached 90%,OGD for 1 h.There was no statistical difference in the cell viability between the model group and the normal group,among the medium/low concentration group of nimodipine injection?three concentration groups of gastrodin injection and the model group(P>0.05);OGD for 3 and 6 h,the cell viability of the model group was significantly lower than the normal group,the medium/low concentration group of nimodipine injection and three concentration groups of gastrodin injection had a protective effect compared with the model group(P<0.05);OGD for 9 h,there was no statistical difference in cell viability between the low concentration group of nimodipine injection and the medium/low concentration group of gastrodin injection compared with the model group(P>0.05),while the other drug groups were lower than the model group(P<0.05).3.After the density of HBMEC reached 90%,OGD for 3 h.Compared with the model group,Claudin-5 and Occludin mRNA transcription and protein expression levels were up-regulated in the nimodipine injection group and gastrodin injection group(P<0.05),but there was no effect on ZO-1(P>0.05).Conclusion:1.At the early stage of OGD,gastrodin injection had an obvious protective effect on HBMEC,but with the extension of OGD time,the protective effect of gastrodin injection disappeared,and the high concentration of drugs would aggravate the cell damage.2.At the early stage of OGD,the protective effect of gastrodin injection on HBMEC and BBB was closely related to the up-regulation of Claudin-5 and Occludin mRNA transcription and protein expression levels.