川陈皮素调节Hippo/YAP信号通路对高糖诱导的人视网膜血管内皮细胞损伤的影响OA

Objective To investigate the effect of nobiletin on high glucose-induced injury of human retinal vascular endothelial cells by adjusting the Hippo/YAP signaling pathway.Methods Human retinal microvascular endothelial cells(HRMEC)were divided into the control group,the high glucose group,the L-nobiletin group,the M-nobiletin group,the H-nobiletin group,and the nobiletin+PY-60 group.HRMECs in the control group were cultured under a glucose concentration of 5.5 mmol·L-1,with 25.0 mmol·L-1 mannitol added to achieve equivalent osmotic pressure.HRMECs in the high glu-cose group,L-nobiletin group,M-nobiletin group,H-nobiletin group,and nobiletin+PY-60 group were cultured in medium con-taining 25.0 mmol·L-1 glucose.After 48 hours,HRMECs in the control and high glucose groups were cultured in normal medium;those in the L-nobiletin,M-nobiletin,and H-nobiletin groups were cultured in medium containing 16 μmol·L-1,32 μmol·L-1,and 64 μmol·L-1 nobiletin,respectively;and those in the nobiletin+PY-60 group were cultured in medium containing 64 μmol·L-1 nobiletin and10 μmol·L-1 PY-60.Cell viability in each group was assessed using the CCK-8 assay;angiogen-esis was observed using Matrigel;cell migration was evaluated using the Transwell assay;reactive oxygen species(ROS)levels were measured using active dihydrorhodamine 123;changes in mitochondrial membrane potential were determined using JC-1 staining;and p-YAP1 protein expression was detected by Western blot.Results Compared with the control group,the high glucose group exhibited decreased HRMEC viability,JC-1 ratio,and p-YAP1 protein level,increased number of angiogenic branches and migrating cells,and elevated ROS fluorescence intensity(all P<0.05).Compared with the high glucose group,the L-nobiletin,M-nobiletin,and H-nobiletin groups showed gradually increased HRMEC viability,JC-1 ratio,and p-YAP1 protein level,gradually decreased number of angiogenic branches and migrating cells,and gradually reduced ROS fluorescence intensity(all P<0.05).Compared with the H-nobiletin group,the nobiletin+PY-60 group showed de-creased relative cell viability of HRMECs,increased number of angiogenic branches,increased number of migrating cells,in-creased relative ROS fluorescence intensity,decreased JC-1 ratio,and decreased level of p-YAP1 protein expression(all P<0.05).Conclusion Nobiletin alleviates high glucose-induced HRMEC injury by activating the Hippo pathway and promo-ting YAP1 phosphorylation.