Mechanism of naringin on retinal microvascular endothelial cells injury based on adenosine-monophosphate-activated protein kinase/mammalian target of rapamycin pathway
- VernacularTitle:基于腺苷酸活化蛋白激酶/哺乳动物雷帕霉素靶蛋白通路探讨柚皮素对视网膜微血管内皮细胞的损伤机制研究
- Author:
Haitong WANG
1
;
Jianliang LIU
Author Information
- Keywords: naringin; retinal microvascular endothelial cells; adenosine-monophosphate-ac-tivated protein kinase/mammalian target of rapamycin pathway; autophagy
- From: Journal of Clinical Medicine in Practice 2024;28(3):23-28
- CountryChina
- Language:Chinese
- Abstract: 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.
