Effect Of Gastrodin On Proliferation And Apoptosis Of Mouse Oligodendrocyte Precursor Cells In Oxygen-glucose Deprivation Model

Objective:To provide theoretical basis for the clinical application of gastrodin in the treatment of white matter demyelination caused by ischemic stroke,we investigated the protective effect of gastrodin on oligodendrocyte precursor cells(OPCs)model of oxygen glucose deprivation(OGD)in vitro.Methods:Experiments were divided into control group and OGD groups conducted within varied time points using primary OPCs from C57BL/6 mouse.The growth curve and cell viability of OPCs were detected by CCK-8,and the proliferation of OPCs under oxygen glucose deprivation and the effect of gastrodin on cell proliferation were detected by icelligence system.The changes of the number of apoptotic cells in the groups,which pretreated with different concentrations of gastrodin(10 ?mol/L,20 ?mol/L,40 ?mol/L),OGD model group and control group at different time points were detected by apoptosis-hoechst staining kit.Choosing the time point(18 h)by the result of CCK-8 and apoptosis staining,when OPCs cell activity and proliferation began to decrease while apoptotic cells began to increase significantly(18 h)as the experimental time point of gastrodin antagonizing OPCs cell proliferation decreasing and apoptosis increasing in OGD model.The diversification of the expression of proliferation-associated protein and apoptosis-related protein were detected by Western Blot and immunofluorescence staining at 18 h.Finally,detecting the regulation of gastrodin on the production of reactive oxygen species(ROS)under the condition of OGD 18 h.Results:?The results of cell growth curve of OPCs showed that the cells of OGD model group increased rapidly at the first 12 h compared with control group,reached the highest at 6 h,then declined gradually,and finally lies behind control group at 18 h(P<0.05).The columnar statistics of cell viability of OPCs showed that the cell viability of OPCs in OGD model group increased before 18 h,and decreased at 18 h compared with control group(P<0.05).? The results of icelligence system showed that the proliferation of OPCs was inhibited under the condition of OGD,and gastrodin could promote the proliferation of OPCs to a certain extent.?Hoechst33258 apoptosis staining showed that the number of apoptotic cells in OGD model group was higher than that in control group,and the increase of apoptotic cells could be reversed by the group treated with gastrodin(10 ?mol/L).? The results of Western Blot and immunofluorescence cytochemical staining showed that gastrodin(10 ?mnol/1)could up-regulate the expression of Cyclin D1(P<0.05)and Ki67(P<0.01)in OPCs under the condition of OGD.At the same time,it could downregulate the expression of cleaved-caspase3 and Bax protein(P<0.01),whilst up-regulate the protein of Bcl-2 and the ratio of Bcl-2/Bax(P<0.01).?The result of ROS detecting showed that gastrodin(10 ?mol/L)could significantly reverse the ROS produced by OPCs cells.Conclusions:The viability of mouse OPCs decreased,the proliferation of OPCs was limited,and the number of apoptotic cells increased under the condition of OGD 18 h simultaneously.Gastrodin could promote the proliferation of OPCs and inhibit the apoptosis of OPCs,which revealed gastrodin can protect OPCs and promote the repair of myelin sheath.and reduce the damage of white matter under cerebral ischemia and hypoxia.