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肾胺酶减轻高糖诱导的足细胞损伤

Remission effects of renalase on high glucose-induced injury in podocytes

  • 摘要:
    目的 探讨肾胺酶对高糖诱导的足细胞损伤的减轻作用及可能机制。
    方法 利用体外培养的小鼠足细胞,分为对照组(5 mmol/L D-葡萄糖)、甘露醇组(5 mmol/L D-葡萄糖+25 mmol/L甘露醇)、高糖组(30 mmol/L D-葡萄糖)与肾胺酶处理组(重组肾胺酶蛋白60 μg/mL预处理6 h+30 mmol/L D-葡萄糖)。48 h后收集细胞,实时定量PCR法测定Ⅳ型胶原α、转化生长因子β1(transforming growth factor β1,TGF-β1)及单核细胞趋化因子1(monocyte chemoattractant protein 1,MCP-1)的mRNA表达水平;酶联免疫吸附试验测定Ⅳ型胶原α、TGF-β1、MCP-1蛋白表达水平、凋亡相关蛋白caspase-3活性和细胞周期蛋白依赖性激酶抑制剂1(cyclin-dependent kinase inhibitor 1,p21)活性。
    结果 与对照组相比,高糖组足细胞Ⅳ型胶原α、TGF-β1、及MCP-1表达水平显著增高,caspase-3活性增加并伴有p21活性增加(P < 0.05);与高糖组相比,肾胺酶处理组足细胞Ⅳ型胶原α、TGF-β1、和MCP-1表达水平下调,caspase-3活性和p21活性下调(P < 0.05)。
    结论 肾胺酶可减轻高糖诱导的足细胞损伤,可能与其抗炎、抗纤维化、抗凋亡和降低p21的活性有关。

     

    Abstract:
    Objective To explore the protective effects and underlying mechanism of renalase on high glucose-induced injury in podocytes.
    Methods Mouse podocytes were cultured in vitro and divided into control group (5 mmol/L glucose), mannitol group (5 mmol/L glucose + 25 mmol/L D-mannitol), high glucose group (30 mmol/L glucose), and study group (60 μg/mL renalase for 6 hours + 30 mmol/L glucose). After cultured for 48 hours, cells were collected. The mRNA levels of collagen Ⅳα, transforming growth factor β1 (TGF-β1), and monocyte chemoattractant protein 1 (MCP-1) were detected by real-time fluorescence quantitative PCR (qRT-PCR). The activity of caspase-3 and cyclin-dependent kinase inhibitor 1 (p21) and levels of collagen Ⅳα, TGF-β1, MCP-1 were detected by enzyme-linked immunosorbent assay.
    Results Compared with the control group, the levels of collagen Ⅳα, TGF-β1, and MCP-1 and the activity of caspase-3 and p21 were significantly increased in podocytes in the high glucose group (P < 0.05). In contrast, the levels of collagen Ⅳα, TGF-β1, and MCP-1, as well as the activity of caspase-3 and p21, were decreased significantly in podocytes in the study group (P < 0.05).
    Conclusions Renalase could attenuate high glucose-induced injury in podocytes, of which the mechanism may be related to anti-inflammatory, anti-fibrotic, anti-apoptotic, and reducing p21 activity.

     

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