Mechanism of naringin on retinal microvascular endothelial cells injury based on adenosine-monophosphate-activated protein kinase/mammalian target of rapamycin pathway
Objective To investigate the mechanism of naringin(NAR)on retinal microvascular endothelial cells(HRMECs)injury based on adenosine-monophosphate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)pathway.Methods HRMECs were randomly di-vided into control group,high glucose(HG)group,HG+NAR group(3 mg/L NAR),HG+activa-tor(AICAR)group(1 mmol/L AICAR),and HG+NAR+AICAR group(3 mg/L NAR+1 mmol/L AICAR);the control group was treated with 5 mmol/L D-glucose added to the culture medium,while the other groups were treated with 30 mmol/L D-glucose added to the culture medium.CCK-8 and Tr-answell were used to detect cell proliferation and migration respectively;enzyme-linked immunosorbent assay(ELISA)was applied to detect the levels of interleukin(IL)-1 β,IL-6 and tumor necrosis factor-α(TNF-α)in the supernatant;quantitative reverse transcriptase polymerase chain reaction(qRT-PCR)was applied to detect the expression levels of autophagy factors LC3 mRNA and p62 mRNA;Western blot was applied to detect the AMPK/mTOR pathway and expression levels of auto-phagy-related proteins.Results Compared with the control group,the cell viability,the number of migrating cells,levels of IL-1,IL-6 and TNF-α,and p-AMPK/AMPK,LC3 Ⅱ/LC3 Ⅰ as well as LC3 mRNA expression increased in the HG group,while the p-mTOR/mTOR,p62 protein and p62 mRNA expression decreased(P＜0.05);compared with the HG group,the cell viability,the num-ber of migrating cells,levels of IL-1,IL-6 and TNF-α,and p-AMPK/AMPK,LC3 Ⅱ/LC3 Ⅰ as well as LC3 mRNA expression decreased in the HG+NAR group,while the p-mTOR/mTOR,p62 protein and p62 mRNA expression increased,but the cell viability,the number of migrating cells,levels of IL-1,IL-6 and TNF-α,p-AMPK/AMPK and LC3 Ⅱ/LC3 Ⅰ expression increased in the HG+AICAR group,while the p-mTOR/mTOR,p62 protein and p62 mRNA expression decreased(P＜0.05);compared with the HG+NAR group,the cell viability,the number of migrating cells,levels of IL-1,IL-6 and TNF-α,and p-AMPK/AMPK,LC3 Ⅱ/LC3 Ⅰ as well as LC3 mRNA ex-pression increased in the HG+NAR+AICAR group,while the p-mTOR/mTOR,p62 protein and p62 mRNA expression decreased(P＜0.05);compared with the HG+AICAR group,the cell via-bility,the number of migrating cells,levels of IL-1,IL-6 and TNF-α,and p-AMPK/AMPK,LC3Ⅱ/LC3 Ⅰ as well as LC3 mRNA expression significantly decreased in the HG+NAR+AICAR group,while the p-mTOR/mTOR,p62 protein and p62 mRNA expression increased(P＜0.05).Conclusion NAR can alleviate HG induced injury to HRMECs,and its mechanism may be related to the inhibition of AMPK/mTOR pathway mediated autophagy.
naringinretinal microvascular endothelial cellsadenosine-monophosphate-ac-tivated protein kinase/mammalian target of rapamycin pathwayautophagy
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