Inhibition Of MiR-143-3p Protects Brain Microvascular Endothelial Cells From Oxygen And Glucose Deprivation Injury

Objective: miRNAs are recognized to play an important role in the neuronal damage and repair in ischemic stroke(IS).The aim of this study is to investigate the effect of inhibitting the expression of miRNA-143-3p on OGD treated BMECs.Methods: qPCR was utilized to measure the expression level of miRNA-143-3 in BMECs after cultured in the condition of OGD.Primary BMECs(p BMECs)of male SD rats were isolated and cultured in vitro to establish ischemic model.p BMECs were divided into four groups: blank control group(p BMEC);positive control group(p BMEC-OGD),p BMECs-NC inhibitor groups(transfected by None senses product inhibitor),p BMECsmiRNA-143-3-inhibitorgroups(transfected by miRNA-143-3-inhibitor).The blank control group was cultured normally,while the other three groups were treated with OGD.miRNA-143-3p expression levels in different groups were examined by qPCR.After that,transwell assays,MTT assays,flow cytometric analysis,and tube formation assays were used to assess the effect of miRNA-143-3p to the cellular damage induced by OGD.Results: The expression of miRNA-143-3p in the BMECs treated with OGD was significantly increased.Within 6 hours of OGD treatment,miRNA-143-3p expression increased rapidly,and then increased slowly.Therefore,6 hours of OGD treatment was selected as the follow-up study time point.Compared to the p BMECs-OGD groups and the p BMECs-NC inhibitor groups,the expression level of miRNA-143-3p was remarkably downregulated in miRNA-143-3p-inhibitor groups.Whereas,no significant difference of the expression of miRNA-143-3p was seen between the blank control groups and the p BMECs-miRNA-143-3p-inhibitor groups.Compared to the p BMECs-NC inhibitor groups and the p BMECs-OGD groups,the miRNA-143-3p-inhibitor groups demonstrated enhancement in cell proliferation,migration and invasion,indicating inhibition of miRNA-143-3p could moderate the OGD-induced cell damage.Parallelly,inhibition of miRNA-143-3p partially abrogated the OGD-induced cell apoptosis and G1 arrest,evidenced by the decease of the cell apoptosis rate and the increase of G2-S1 ration in miRNA-143-3p-inhibitor groups in comparison with other two OGD-treated groups.In addition,miRNA-143-3p-inhibitor groups presented upregulation of tube formation compared to others OGD groups.Conclusions: Inhibition of miRNA-143-3p expression can significantly alleviate the damage of BMECs induced by OGD and play a cytoprotective role.Inhibitting the expression of miRNA-143-3p may be a potential therapeutic target for ischemic stroke.