阿霉素致足细胞损伤模型的建立

Establishment of the Adriamycin-induced Injury Model of Podocyte Cells

目的建立一种稳定的小鼠足细胞损伤模型，为进一步研究足细胞的生物特性，以及其相关蛋白与肾性蛋白尿间的关系提供可靠保证。方法应用CCK8检测不同浓度的阿霉素（0．5μg／ml，0．25μg/ml，0．125μg/ml）分别作用24h，48h后，足细胞的抑制率；应用流式细胞术检测测定足细胞凋亡情况;应用R—TPCR检测裂孔膜关键因子nephrin，podocin的表达。结果0．25μg／ml的阿霉素作用24h，抑制率接近50％；流式细胞术及R—TPCR均表明浓度为0．25μ/ml的ADR作用24h后与正常组存在显著性差异。结论0．25μg／ml的阿霉素作用24h建立的足细胞模型稳定可靠，可应用于足细胞研究。

Objective To establish a stable podocyte injury model to further study biological features of podocytes, and the relationship between podocyte-related proteins and renal proteinuria. Methods Podocytes were treated for 24 and 48 h with 0.5, 0.25, 0. 125 μg/mL adriamycin （ADR）, respectively. The inhibition rate of podocytes was detected by using CCK8. Flow cytometry was used to detect the apoptosis of podocyte cells, and R-T PCR to measure the expression of nephrin and podocin. Results The inhibition rate was nearly 50 % in podocytes treated with 0. 25 μg/ mL ADR for 24 h. Flow cytometry and R-T PCR showed that there were significant differences in the apoptosis of podocytes and expression levels of nephrin and podocin between 0. 25μg/mL ADR group and normal group （ P 〈 0. 05 ） . Conclusion The podoeyte injury model induced by 0.25μg/mL ADR is stable and reliable, which can be used for further studies on podocyte injury.