靶向lncRNA H19/miR-19b轴探究Riligustilide在高糖诱导人视网膜微血管内皮细胞炎症中的调控机制OA

Objective To investigate the effect of Riligustilide on high glucose-induced injury in human retinal micro-vascular endothelial cells(HRMECs)and to determine whether this effect is mediated through regulation of the long non-coding RNA H19(lncRNA H19)/microRNA-19b(miR-19b)axis.Methods HRMECs were divided into a normal glucose group(NG group),a mannitol control group(Man group),a high glucose group(HG group),a low-dose Riligustilide group(HG+Rili-L group),and a high-dose Riligustilide group(HG+Rili-H group).Cell viability(OD450)was assessed using the Cell Counting Kit-8(CCK-8)assay.The relative mRNA expression levels of interleukin(IL)-6,IL-1β,tumor necrosis factor-α(TNF-α),lncRNA H19,and miR-19b were detected by quantitative real-time polymerase chain reaction(qRT-PCR).Oxi-dative stress was evaluated using the superoxide dismutase(SOD),lactate dehydrogenase(LDH),and malondialdehyde(MDA)detection kit.Immunofluorescence staining was used to detect the expression of occludin and zonula occludens-1(ZO-1).A dual-luciferase reporter gene assay was performed to verify the targeting relationship between lncRNA H19 and miR-19b.Small interfering RNA targeting H19(si-H19)was transfected to determine the effect of H19 silencing on the ac-tion of Riligustilide.Results The CCK-8 assay showed that low-and moderate-concentration Riligustilide had no signifi-cant effect on the viability of HRMECs,whereas 100 μmol·L-1 Riligustilide reduced cell viability;therefore,5 μmol·L-1 and 10 μmol·L-1 were selected for subsequent experiments.Compared with the NG group,no significant differences were observed among the indicators in the Man group(all P＞0.05);however,in the HG group,OD450 value and SOD activity de-creased,the relative mRNA expression levels of IL-6,IL-1β,and TNF-α,as well as the LDH level and MDA content in-creased,and the expression of occludin and ZO-1 were reduced(all P＜0.05).Compared with the HG group,both low-and high-dose Riligustilide significantly increased cell viability,downregulated inflammatory cytokine expression,alleviated oxi-dative stress injury,and enhanced the expression of occludin and ZO-1,with the high-dose Riligustilide showing a more pro-nounced effect(all P＜0.05).The dual-luciferase reporter gene assay confirmed that lncRNA H19 could bind to miR-19b.qRT-PCR results showed that lncRNA H19 expression was downregulated and miR-19b expression was upregulated in the HG group,whereas Riligustilide treatment increased the lncRNA H19 expression and decreased the miR-19b expression(all P＜0.05).H19 silencing partially reversed the inhibitory effects of Riligustilide on inflammatory response and oxidative stress injury.Conclusion Riligustilide alleviates high-glucose-induced injury in HRMECs by regulating the lncRNA H19/miR-19b axis.