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目的:探讨高糖对小鼠足细胞上皮间充质转分化(EMT)与迁移的影响及其分子机制。方法:以永生化小鼠足细胞株(足细胞)为研究对象,设置正常糖组(5 mmol/L葡萄糖)、甘露醇组(5 mmol/L葡萄糖+25mmol/L甘露醇)、高糖组(30 mmol/L葡萄糖)、核因子κB(NF-κB)特异性抑制剂小白菊内酯(PTL)+高糖组,作用时间24h。采用Transwell检测细胞迁移能力,Western Blot检测蛋白表达,q PCR检测mRNA表达情况,免疫荧光检测NF-κB入核情况。结果:与正常糖及甘露醇组比较,高糖刺激足细胞24h后细胞的迁移能力明显增强(P<0.05),足细胞特征性蛋白nephrin明显下调(P<0.05),纤连蛋白(FN)及mRNA表达显著增加(P<0.05),而E钙黏蛋白(E-cadherin)及其mRNA表达明显下降(P<0.05)。同时,与上述对照组比较,高糖明显上调p-IκBα(P<0.05)下调IκBα(P<0.05)促使足细胞胞质中的NF-κB入核同时上调p-p65的表达;干预细胞PTL后,足细胞的EMT和迁移得到明显抑制(P<0.05),nephrin蛋白表达明显上升(P<0.05)。结论:高糖通过激活NF-κB通路诱导足细胞发生EMT及迁移。
Objective: To investigate the effect and mechanism of high glucose on epidermal mesenchymal transition (EMT) and migration in mouse podocytes. Methods: The immortalized mouse podocyte (podocyte) was used as research object. Normal glucose group (5 mmol / L glucose), mannitol group (5 mmol / L glucose + 25 mmol / L mannitol) (30 mmol / L glucose), and NF-κB-specific inhibitor of parthenolide (PTL) + high glucose for 24h. Transwell assay was used to detect cell migration. Western Blot was used to detect the protein expression. Q-PCR was used to detect mRNA expression. Immunofluorescence was used to detect the nuclear translocation of NF-κB. Results: Compared with normal glucose and mannitol groups, the migration ability of podocytes was significantly increased (P <0.05), podocyte nephrin (P <0.05), fibronectin (FN) (P <0.05), while the expression of E-cadherin and its mRNA was significantly decreased (P <0.05). At the same time, compared with the above control group, high-glucose significantly increased p-IκBα (P <0.05) downregulation of IκBα (P <0.05), podocyte nuclear factor- (P <0.05). The nephrin protein expression was significantly increased (P <0.05). Conclusion: High glucose induces EMT and migration of podocytes by activating NF-κB pathway.