摘要
目的探讨衰老相关肾间质纤维化的肾缺氧特点以及与微小RNA(microRNA,miRNA)表达谱特征的联系,并验证尿液外泌体miRNA作为非侵入性生物标志物的潜力。方法选择自然衰老大鼠(24月龄)与年轻大鼠(1月龄)模型,通过免疫组化、Masson染色检测两组大鼠肾组织肾间质纤维化、胶原沉积以及缺氧情况,同时通过western blot检测两组肾组织缺氧标志物缺氧诱导因子1α(hypoxia inducible factor-1α,HIF-1α)、间质纤维化指标[E-钙黏蛋白(epithelial cadherin,E-cadherin)、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)]表达情况。通过GO分析和KEGG pathway分析,筛选出受HIF-1α调控,且两组表达差异较大的miRNA,使用实时定量PCR在两组肾组织以及成人尿液外泌体中进行验证。结果衰老大鼠肾组织呈现显著缺氧(HIF-1α表达升高)和间质纤维化(胶原沉积增加、α-SMA表达增加,E-cadherin表达下降),且伴随miR-21(2.49倍)、miR-34a(2.35倍)显著升高及miR-23a(0.46倍)、miR-210(0.38倍)、miR-205(0.51倍)显著降低,差异有统计学意义(P均<0.01)。进一步通过成人尿液外泌体验证,发现相比较年轻组,衰老组成人尿液外泌体中miR-21(5.73倍)、miR-34a(2.05倍)水平显著升高,miR-23a(0.42倍)水平降低,差异有统计学意义(P<0.05)。结论miR-21、miR-34a上调与miR-23a下调可能通过缺氧途径驱动参与衰老相关肾间质纤维化,可作为非侵入性诊断标志物及潜在的治疗靶点。
Objective To examine the characteristics of renal hypoxia and the microRNA(miRNA)expression profile associated with aging-related renal interstitial fibrosis,as well as to evaluate the potential of urinary exosomal miRNA as a non-invasive biomarker.Methods In this study,naturally aged rats(24 months old)and young rats(1 month old)were utilized as experimental models.Renal interstitial fibrosis,collagen deposition,and hypoxia within the renal tissues of both groups were assessed using immunohistochemistry and Masson staining techniques.The expression levels of hypoxia markers[hypoxia inducible factor-1α(HIF-1α)]and interstitial fibrosis indicators[epithelial cadherin(E-cadherin),α-smooth muscle actin(α-SMA)] in the renal tissues were quantified via western blot analysis.GO and KEGG pathway analyses were conducted to identify miRNAs regulated by HIF-1α that exhibited significant differential expression between the two groups.The findings were further validated through real-time quantitative PCR in both the renal tissues and the exosomes derived from adult urine.ResultsAging rat renal tissue exhibited notable hypoxia(elevated HIF-1α expression)and interstitial fibrosis(higher collagen deposition,increased α-SMA expression,and reduced E-cadherin expression).These changes were accompanied by a marked upregulation of miR-21(2.49-fold increase)and miR-34a(2.35-fold increase),and a significant downregulation of miR-23a(0.46-fold decrease),miR-210(0.38-fold decrease),and miR-205(0.51-fold decrease),and the differences were statistically significant(P<0.01).Further validation using adult urine exosomes revealed that,in comparison to the young group,the aged group demonstrated significantly elevated levels of miR-21(5.73-fold increase)and miR-34a(2.05-fold increase),along with a reduced level of miR-23a(0.42-fold decrease),and the differences were statistically significant(P<0.05).Conclusion The upregulation of miR-21 and miR-34a,along with the downregulation of miR-23a,may contribute to aging-related renal interstitial fibrosis via the hypoxia pathway.These miRNAs hold promise as non-invasive diagnostic markers and potential therapeutic targets.
作者
刘金瑞
张靖华
杨青彦
武相
王长安
LIU Jin-rui;ZHANG Jing-hua;YANG Qing-yan;WU Xiang;WANG Chang-an(Department of Kideny,Transplantion and Nephrology,the 7th Peoples Hospital of Zhengzhou,Zhengzhou,Henan 450016,China)
出处
《医药论坛杂志》
2025年第18期1926-1931,共6页
Journal of Medical Forum
基金
河南省科技攻关计划项目(232102310490,242102311081)。
