间充质干细胞外泌体lncRNA ZEB1-AS1通过靶向miR-142-5p/PTEN调控糖尿病性肾病

Exosomal lncRNA ZEB1-AS1 secreted from mesenchymal stem cells attenuates diabetic nephropathy by regulating miR-142-5p/PTEN axis

目的探讨脂肪源间充质干细胞(adMSC)的外泌体长链非编码RNA锌指E盒结合同源框1反义链1(Zinc finger E-box binding homeobox 1 antisense 1,ZEB1-AS1)在糖尿病肾病(DN)中的作用机制。方法采用大鼠DN模型和高糖(HG)诱导的肾小球系膜细胞(GMCs)模型进行生化分析和炎症/氧化应激检测;双荧光素酶基因报告实验验证ZEB1-AS1、第10号染色体同源丢失性磷酸酶-张力蛋白基因(PTEN)与microRNA(miR)-142-5p的结合关系;Western blotting检测PTEN蛋白表达水平。结果adMSC分泌的外泌体ZEB1-AS1可降低DN大鼠血糖、血清肌酐、24 h尿蛋白、肾脏重量、肾组织纤维化以及炎症细胞浸润水平。同时,在DN大鼠和HG诱导的GMCs中,肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和白细胞介素-1β(IL-1β)的表达下降,而谷胱甘肽过氧化物酶(GPx)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽(GSH)的表达上升。此外,miR-142-5p能与ZEB1-AS1、PTEN结合;miR-142-5p过表达或PTEN沉默逆转了外泌体ZEB1-AS1对炎症细胞因子水平的抑制作用和对抗氧化酶浓度的促进作用。结论来自adMSC的外泌体ZEB1-AS1通过miR-142-5p/PTEN通路,控制炎症和氧化应激来抑制DN的进展。这表明ZEB1-AS1可能是治疗DN潜在、有效的靶点。

Objective To explore the role of an exosomal long non-coding RNA zinc finger E-box-bingding homeobox 1 antisense 1(ZEB1-AS1)from adipose-derived MSC(adMSC)and its action mechanism in DN.Methods DN rat model and high glucose(HG)-induced glomerular mesangial cell(GMCs)model treated with exosomal ZEB1-AS1 from adMSC were used for biochemical analysis and inflammation/oxidative stress assessment.The binding relationships among ZEB1-AS1,microRNA(miR)-142-5p,and phosphatase and tensin homolog deleted on chromosome 10(PTEN)were confirmed by dual luciferase reporter assay.Western blotting was used for the measurement of PTEN protein level.Results adMSC-secreted exosomal ZEB1-AS1 reduced the levels of blood glucose,serum creatinine,24 h urinary protein,kidney weight,fibrosis,and inflammatory cell infiltrations in DN rats.Meanwhile,both in DN rats and HG-induced GMCs,the levels of tumor necrosis factor(TNF)-α,interleukin(IL)-6,and IL-1βwere inhibited,but glutathione peroxidase(GPx),superoxide dismutase(SOD),catalase(CAT),and glutathione(GSH)were promoted by exosomal ZEB1-AS1 treatment.Additionally,miR-142-5p was identified to bind to ZEB1-AS1,while miR-142-5p further targeted PTEN.miR-142-5p overexpression or PTEN silencing reversed the inhibiting effects of exosomal ZEB1-AS1 on inflammatory cytokine levels and the promoting effects on the concentrations of antioxidant enzymes.Conclusion Exosomal ZEB1-AS1 from adMSC prevents DN progression by controlling inflammation and oxidative stress via the miR-142-5p/PTEN pathway,suggesting that ZEB1-AS1 may serve as a potential and effective therapeutic target for treating DN.