Slit引导配体2通过调控AMPK/SIRT1-FoxO1信号通路影响糖尿病小鼠视网膜血管损伤的机制研究  

ffStudy on the mechanism of Slit guiding ligand 2 affecting retinal vascular injury in diabetic mice by regulating AMPK/SIRT1-FoxO1 signaling pathway

作　　者：李天航 顾朝辉[1] 张月玲[1] 李洁[1] 杜鹃 付燕[1] 陈娜[1] 陈佳菲 LI Tianhang;GU Zhaohui;ZHANG Yueling;LI Jie;DU Juan;FU Yan;CHEN Na;CHEN Jiafei(The Second Department of Ophthalmology,Baoding No.1 Central Hospital,Baoding 071000,China)

机构地区：[1]河北省保定市第一中心医院眼2科,河北保定071000

出　　处：《长春中医药大学学报》2024年第11期1214-1219,共6页Journal of Changchun University of Chinese Medicine

基　　金：河北省2022年度医学科技研究课题(20220293)。

摘　　要：目的 探讨Slit引导配体2(SLIT2)是否通过调控腺苷单磷酸活化蛋白激酶(AMPK)/转录沉默信息调节因子1(SIRT1)-叉头盒蛋白O1(FoxO1)信号通路通过对糖尿病小鼠视网膜血管损伤的影响。方法 30只db/db小鼠随机分为DR组(db/db小鼠)、DR+阴性对照载体组(DR+sh-NC组)和DR+sh-SLIT2组,每组10只。另选10只db/m小鼠为对照组。DR+sh-NC组和DR+sh-SLIT2组麻醉后分别在双眼玻璃体腔内注射sh-SLIT2的腺相关病毒(AAV)载体。眼底荧光血管造影(FFA)和苏木精伊红染色观察视网膜血管病变;酶联免疫吸附测定检测血清白细胞介素-6(IL-6),肿瘤坏死因子α(TNF-α)和血管内皮生长因子(VEGF)的水平,荧光定量PCR检测SLIT2 mRNA表达;Western blot检测视网膜组织SLIT2、AMPK、SIRT1、FoxO1蛋白水平。结果 与对照组相比,DR组、DR+sh-NC组、DR+sh-SLIT2组血糖、每日饮水量、每日排尿量、食物摄入量及体质量均明显升高(P<0.05);与对照组相比,DR组视网膜存在血管病变及病理损伤,SLIT2 mRNA及蛋白表达、IL-6、TNF-α和VEGF水平,FoxO1蛋白水平均明显升高(P<0.05),AMPK、SIRT1蛋白水平均明显降低(P<0.05);与DR+sh-NC组相比,DR+sh-SLIT2组的视网膜血管病变及病理损伤明显减轻,SLIT2 mRNA及蛋白表达、IL-6、TNF-α和VEGF水平,FoxO1蛋白水平均明显降低(P<0.05),AMPK、SIRT1蛋白水平均明显升高(P<0.05)。结论 沉默SLIT2表达显著改善糖尿病小鼠视网膜血管损伤及炎症水平,这可能是通过调控AMPK/SIRT1-FoxO1信号通路发挥作用的。Objective To investigate the effect of Slit guiding ligand 2(SLIT2)on retinal vascular injury in diabetic mice by regulating the adenosine monophosphate activated protein kinase(AMPK)/silent information regulator of transcription 1(SIRT1)-forkhead box protein O1(FoxO1)signaling pathway.Methods 30 db/db mice were randomly divided into diabetic retinopathy(DR)group(db/db mice),DR+negative control carrier group(DR+sh-NC)and DR+sh-SLIT2 group,with 10 mice in each group.Another 10 db/m mice were selected as control group.DR+sh-NC group and DR+sh-SLIT2 group were injected with adeno-associated virus(AAV)vector containing sh-SLIT2 into the vitreous cavities of both eyes after anesthesia.Retinal vascular lesions were observed by fundus fluorescence angiography(FFA)and hematoxylin eosin staining.Serum interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and vascular endothelial growth factor(VEGF)levels were detected by enzyme linked immunosorbent assay(ELISA),and SLIT2 mRNA expression was detected by fluorescence quantitative real-time PCR.The protein levels of SLIT2,AMPK,SIRT1 and FoxO1 in retinal tissue were detected by Western blot.Results Compared with the control group,blood glucose level,daily water intake,daily urine output,food intake and body weight in DR group,DR+sh-NC group and DR+sh-SLIT2 group were significantly increased(P<0.05).Compared with the control group,the DR group showed vascular lesions and pathological damage in the retina,with significant increases in SLIT2 mRNA and protein expression,IL-6,TNF-αand VEGF levels,and FoxO1 protein levels(P<0.05),and significant decreases in AMPK and SIRT1 protein levels(P<0.05).Compared with the DR+sh-NC group,the retinal vascular lesions and pathological damage in DR+sh-SLIT2 group were significantly reduced,SLIT2 mRNA and protein expression,IL-6,TNF-αand VEGF levels,and FoxO1 protein levels were significantly decreased(P<0.05),while the protein levels of AMPK and SIRT1 were significantly increased(P<0.05).Conclusion Silencing SLIT2 expression significantly impro

关 键 词：Slit引导配体2 腺苷单磷酸活化蛋白激酶 转录沉默信息调节因子1 叉头盒蛋白O1 糖尿病视网膜病变 

分 类 号：R774.1[医药卫生—眼科]