山西医科大学学报2025，Vol.56Issue(2)：165-174,10.DOI:10.13753/j.issn.1007-6611.2025.02.008
FLCN对糖尿病视网膜病变发生发展的影响及其机制
Effect and mechanism of FLCN on the occurrence and development of diabetic retinopathy
摘要
Abstract
Objective To investigate the effects of folliculin(FLCN)on the occurrence and development of diabetic retinopathy(DR)and AMP-activated protein kinase(AMPK)signaling pathway.Methods The human retinal pigment epithelial cells(ARPE-19)were divided into control group,high glucose group,shRNA control(NC-sh)group,FLCN-shRNA(FLCN-sh)group,overexpression control(NC-oe)group,and FLCN overexpression(FLCN-oe)group.After transfection,the cells in control group were cultured in me-dium containing normal glucose(5 mmol/L)for 48 h,and the cells in the other groups were cultured in medium containing high glucose(25 mmol/L)for 48 h.Cell viability was detected by CCK-8,and cell proliferation was detected by EdU methods.Cell apoptosis was detected by TUNEL method.The diabetic rat model was established by one-time intraperitoneal injection of 1%streptozotocin(STZ)solution(35 mg/kg),and the diabetic rats were fed for one month to establish the DR model.After modeling,DR model rats were di-vided into DR group,DR+NC-sh group and DR+FLCN-sh group,with 12 rats in each group.Another 12 normal rats were chosen as control group.The rats in control group and DR group were injected with 20 μL of normal saline into the vitreous cavity,and the rats in DR+NC-sh group and DR+FLCN-sh group were injected with 20 μL of NC shRNA(1×109 TU/mL)and FLCN shRNA(1×109 TU/mL)into the vitreous cavity,respectively.All rats were injected once every seven days for four times.The retinal morphology was observed by HE staining,and the levels of oxidative stress indexes[superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-Px),and malondialdehyde(MDA)]and inflammation indicators(TNF-α,IL-1β,and IL-6)in ARPE-19 cells and rat retinal tissues were detected according to the instructions of the kit.The levels of FLCN mRNA in ARPE-19 cells and rat retinal tissues were detected by qRT-PCR.The protein levels of FLCN,p-AMPKα,AMPKα,mitochondrial transcription factor A(TFAM),mitochondrial fusion protein 1(MFN1)and mitochondrial fusion protein 2(MFN2)in ARPE-19 cells and rat retinal tissues were detected by Western blot.Results ①Compared with high glucose group and NC-sh group,FLCN mRNA and protein levels in ARPE-19 cells were decreased in FLCN-sh group(P<0.05),the relative cell viability and EdU positive rate were increased(P<0.05),the cell apoptosis was decreased(P<0.05),SOD,CAT and GSH-Px levels were increased(P<0.05),MDA,TNF-α,IL-1β and IL-6 levels were decreased(P<0.05),and the protein levels of P-AMPKα,TFAM,MFN1 and MFN2 were increased(P<0.05).Compared with high glucose group and NC-oe group,FLCN mRNA and protein levels were increased in FLCN-oe group(P<0.05),the cell viability and EdU positive rate were decreased(P<0.05),the cell apoptosis was increased(P<0.05),SOD,CAT and GSH-Px levels were decreased(P<0.05),MDA,TNF-α,IL-1β and IL-6 levels were increased(P<0.05),and the protein levels of p-AMPKα,TFAM,MFN1 and MFN2 were decreased(P<0.05).②Compared with DR group and DR+NC-sh group,FLCN mRNA and protein levels in retinal tissues were decreased in DR+FLCN-sh group(P<0.05),the fundus neovascularization was significantly reduced,the retinal morphology was significantly improved,SOD,CAT and GSH-Px levels in retinal tissues were increased(P<0.05),the levels of MDA,TNF-α,IL-1β and IL-6 were decreased(P<0.05),and the protein levels of p-AMPKα,TFAM,MFN1 and MFN2 in retinal tissue were increased(P<0.05).Conclusion Downregulation of FLCN can alleviate high glucose-induced ARPE-19 cell damage and retinal damage in DR rats by activating the AMPK signaling pathway.关键词
糖尿病视网膜病变/卵泡蛋白/腺苷酸活化蛋白激酶/视网膜色素上皮/氧化应激/炎症Key words
diabetic retinopathy/folliculin/adenylate activated protein kinase/retinal pigment epithelium/oxidative stress/inflammation分类
临床医学引用本文复制引用
赵吉飞,张婕,刘甜,陈宁宁,王莉,秦扬,杨永明..FLCN对糖尿病视网膜病变发生发展的影响及其机制[J].山西医科大学学报,2025,56(2):165-174,10.基金项目
陕西省重点研发计划-国际科技合作计划项目(2022kw-43) （2022kw-43）
咸阳市重点研发计划项目(L2022ZDYFSF038) （L2022ZDYFSF038）
