黄芪多糖对小鼠肾小球足细胞Nephrin、Desmin表达的影响

Effect of Astragalus Polysaccharide on Expression of Nephrin and Desmin in Mouse Glomerular Podocytes

目的:观察黄芪多糖(astragalus polysaccharide,APS)对阿霉素(adriacin doxorubicin,ADR)诱导的肾小球足细胞损伤中足细胞裂孔隔膜蛋白Nephrin及骨架蛋白Desmin表达的影响。方法:通过体外培养小鼠肾小球足细胞,以ADR诱导制作足细胞损伤模型,分为对照组、ADR损伤组(0.5 mg·L^-1 ADR)、APS干预组(0.5 mg·L^-1 ADR+0.8 g·L^-1APS)。MTT比色法检测3组细胞活力;免疫荧光染色观察3组Nephrin、Desmin表达;Western blot检测Nephrin、Desmin蛋白表达;RT-PCR检测Nephrin、Desmin mRNA水平。结果:APS干预组、对照组细胞活力[(87.25±11.14)%、(97.05±12.67)%]高于ADR损伤组[(58.62±7.93)%](P<0.05)。免疫荧光显示:APS干预组足细胞Nephrin表达少于对照组,但明显强于ADR损伤组(P<0.05);而足细胞Desmin少量表达,且明显少于ADR损伤组(P<0.05)。对照组、APS干预组足细胞Nephrin蛋白含量及Nephrin mRNA水平高于ADR损伤组(P<0.05),Desmin蛋白含量及Nephrin mRNA水平低于ADR损伤组,差异有统计学意义(P<0.05);而APS干预组与对照组足细胞Nephrin、Desmin蛋白及Nephrin、Desmin mRNA表达比较,差异无统计学意义(P>0.05)。结论:黄芪多糖对ADR诱导的肾小球足细胞损伤具有保护作用,其机制可能与上调Nephrin表达、下调Desmin表达有关。

Objectives:To observe the effect of astragalus polysaccharide(APS) on the expression of nephrin and skeletal protein Desmin in the podocyte injury of glomerular podocytes induced by adriacin doxorubicin(ADR).Methods:The rat glomerular podocytes were cultured in vitro,and the podocyte injury model was induced by ADR.The model was divided into control group,ADR injury group(0.5 mg·L^-1 ADR) and APS intervention group(0.5 mg·L^-1 ADR +0.8 g·L^-1 APS).The viability of three groups was detected by MTT colorimetric assay.The expressions of nephrin and Desmin were detected by immunofluorescence staining.The expressions of nephrin and Desmin were detected by Western blot.The levels of nephrin and Desmin mRNA were detected by RT-PCR.Results:The cell viability of the APS intervention group and the control group [(87.25±11.14)%,(97.05±12.67)%] was higher than that of the ADR injury group [(58.62±7.93)%](P<0.05).Immunofluorescence showed that the expression of nephrin in the APS intervention group was less than that in the control group,but it was significantly higher than that in the ADR injury group(P<0.05).The expression of Desmin in the podocytes was significantly lower than that in the ADR injury group(P<0.05).The levels of Nephrin protein and Nephrin mRNA in the control group and APS intervention group were higher than those in ADR injury group(P<0.05),Desmin protein content and Nephrin mRNA level were lower than those in ADR injury group(P<0.05).There was no significant difference in the expression of nephrin,Desmin protein and Nephrin and Desmin mRNA between APS intervention group and control group(P>0.05).Conclusion:Astragalus polysaccharide has protective effect on ADR-induced glomerular podocyte injury,and its mechanism may be related to up-regulation of nephrin expression and down-regulation of Desmin expression.