Effects of LOXL1-AS1 on oxygen-glucose deprivation-induced injury of human brain microvascular endothelial cells / 中风与神经疾病杂志

ABSTRACT

@#Objective To investigate the effects of the long non-coding RNA LOXL1 antisense RNA 1 （LOXL1-AS1） on the apoptosis and inflammatory factor expression of human brain microvascular endothelial cells （HBMECs） induced by oxygen-glucose deprivation （OGD）. Methods HBMECs were divided into control group （normal culture）， OGD group （OGD injury）， OGD+si-NC group （transfection with si-NC plus OGD injury）， OGD+si-LOXL1-AS1 group （transfection with si-LOXL1-AS1 plus OGD injury）， OGD+miR-NC group （transfection with miR-NC plus OGD injury）， OGD+miR-761 group （transfection with miR-761 mimic plus OGD injury）， OGD+si-LOXL1-AS1+negative control group （transfection with si-LOXL1-AS1 and anti-miR-NC plus OGD injury）， and OGD+si-LOXL1-AS1+miR-761 inhibitor group （transfection with si-LOXL1-AS1 and miR-761 inhibitor plus OGD injury）. The expression of LOXL1-AS1 and miR-761 was measured by RT-qPCR. Cell viability was measured using cell counting kit-8. Cell apoptosis was determined by flow cytometry. The expression of B-cell lymphoma/leukemia-2 （Bcl-2） protein and Bcl-2-associated X （Bax） protein was measured by Western blotting. The levels of interleukin （IL）-6， IL-1β， and tumor necrosis factor-α （TNF-α） were measured by enzyme-linked immunosorbent assay. Dual luciferase reporter assay was used to detect the complementary binding of LOXL1-AS1 and miR-761. Results Compared with the control group， the OGD group showed significant increases in LOXL1-AS1 expression， the cell apoptosis rate， Bax expression， and IL-6， IL-1β， and TNF-α levels and significant decreases in the cell survival rate and Bcl-2 expression （all P<0.05）. After inhibiting LOXL1-AS1， the OGD+si-LOXL1-AS1 group showed significant decreases in LOXL1-AS1 expression， the apoptosis rate， Bax expression， and IL-6， IL-1β， and TNF-α levels and significant decreases in the survival rate and Bcl-2 expression， compared with the OGD group and the OGD+si-NC group （all P<0.05）. LOXL1-AS1 targeted the expression of miR-761. After overexpressing miR-761， the OGD+miR-761 group showed significant increases in the survival rate and Bcl-2 expression and significant decreases in the apoptosis rate， Bax expression， and IL-6， IL-1β， and TNF-α levels， compared with the OGD+miR-NC group （all P<0.05）. Compared with the OGD+si-LOXL1-AS1+negative control group， the OGD+si-LOXL1-AS1+miR-761 inhibitor group showed significantly decreased survival rate and Bcl-2 expression and significantly increased apoptosis rate， Bax expression， and IL-6， IL-1β， and TNF-α levels （all P<0.05）. Conclusion Inhibiting LOXL1-AS1 expression can up-regulate miR-761 to promote the survival of OGD-induced HBMECs and suppress the cells' apoptosis and expression of inflammatory factors.