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目的观察晚期糖基化终产物(advanced glycation end-products,AGEs)对足细胞synaptopodin的影响和在足细胞迁移性中的作用,探讨其作用机制。方法不同浓度AGEs(0、20、40、80μg/mL)干预足细胞24 h,免疫印迹检测足细胞synaptopodin的表达,激光共聚焦显微镜观察足细胞肌动蛋白F-actin和synaptopodin的分布,Transwell细胞迁移实验检测足细胞的迁移性,氯沙坦(100μmol/L)预处理足细胞后,观察synaptopodin和F-actin的改变。结果 AGEs 80μg/mL可降低足细胞synaptopodin的表达[(50±11)%vs 100%,P<0.05],并使肌动蛋白骨架F-actin发生重构,足细胞的迁移性增高[(47±6)vs(13±3),P<0.05],氯沙坦可减轻AGEs介导的synaptopodin的下调[(80±12)%vs(50±11)%,P<0.05]和F-actin的重构,降低足细胞的迁移性[(25±4)vs(47±6),P<0.05]。结论 AGEs可能通过激活足细胞内的肾素-血管紧张素系统的机制,下调synaptopodin表达,介导骨架蛋白的重构,增加迁移性。
Objective To investigate the effects of advanced glycation end-products (AGEs) on podocyte synaptopodin and its role in podocyte migration and to explore its mechanism. Methods Different concentrations of AGEs (0, 20, 40 and 80 μg / mL) were used to treat podocytes for 24 h. The expression of synaptopodin in podocytes was detected by immunoblotting. The distribution of actin F-actin and synaptopodin in podocytes was observed by laser confocal microscopy. Migration assay was used to detect the migration of podocytes. After pretreatment of podocytes with Losartan (100μmol / L), changes of synaptopodin and F-actin were observed. Results AGEs 80μg / mL decreased the expression of synaptopodin in podocytes [(50 ± 11)% vs 100%, P <0.05], and remodeled actin cytoskeleton F-actin Losartan reduced AGEs-mediated down-regulation of synaptopodin [(80 ± 12)% vs (50 ± 11)%, P <0.05] and F-actin The remodeling of podocytes reduced the migration of podocytes [(25 ± 4) vs (47 ± 6), P <0.05]. Conclusion AGEs may down-regulate the expression of synaptopodin by activating the renin-angiotensin system in podocytes, mediate the remodeling of skeletal proteins and increase the mobility of AGEs.