五味子醇提液对阿霉素损伤足细胞中nephrin和desmin表达的影响

Effects of Schisandra Chinensis Fruit Ethanol Extract on Nephrin and Desmin Expression in Adriamycin Induced Podocyte Injury

目的观察五味子醇提液(SE)对阿霉素(ADR)诱导的体外足细胞损伤中足细胞裂孔膜蛋白nephrin和骨架蛋白desmin表达的影响,并探讨其作用机制。方法体外ADR作用于足细胞24 h,造成足细胞损伤后,加入含SE的培养基,继培养24 h。设对照组(0 mg/L ADR)、模型组(0.25 mg/L ADR)、SE干预组(250 mg/L SE+0.25 mg/LADR)、SE组(250 mg/L SE)。免疫荧光法测定各组nephrin的表达;Western Blot检测各组nephrin和desmin蛋白表达;RT-PCR检测各组nephrin和desmin mRNA表达。结果免疫荧光结果显示对照组和SE组nephrin呈颗粒状或线性分布于细胞膜,模型组较对照组、SE组和SE干预组表达强度明显减少。模型组nephrin蛋白和mRNA表达较对照组显著减少,SE干预组nephrin蛋白和mRNA表达水平较模型组增加。模型组desmin蛋白和mRNA表达较对照组显著增加,SE干预组desmin蛋白和mRNA表达水平较模型组显著减少(均P<0.05)。SE和SE干预组的nephrin和desmin蛋白和mRNA表达水平与对照组差异无统计学意义。结论 250 mg/L SE对体外培养足细胞无损伤作用,且SE能拮抗ADR对足细胞的损伤,这可能与SE可以上调nephrin和下调desmin的表达有关。

Objective To explore the effect of schisandra chinensis fruit ethanol extract on nephrin and desmin ex-pression in adriamycin（ADR） induced podocyte injury in vitro. Methods Conditionally immortalized mouse podocytes were treated with ADR for 24 h in vitro, then the medium was changed to medium with SE（250 mg/L）for 24 h. Podocytes were di-vided into four groups：control group,model group, SE intervention group and SE group. The expression of nephrin in podo-cytes was detected by immunofluorescence. Western Blot was employed to assess nephrin and desmin expression. Transcrip-tion level of nephrin and desmin were determined by qRT-PCR. Results Nephrin expression was distributed along the cell membrane in linear or granular pattern in control group and SE group. Fluorescence intensity in model group was lower than that of control group SE group and SE intervention group. There was no significant difference of nephrin and desmin protein and mRNA level between control group and SE group. Compared with the model group, protein and mRNA level of nephrin was lower than that of control group and SE intervention group. The protein expression and mRNA transcription of desmin in model group was higher than those in control group and SE intervention group （P〈0.05）. Conclusion SE（250 mg/L）has no harmful effect on the podocytes in vitro. SE can protect the podocytes from damage by adriamycin in vitro. SE not only up-regulate the expression of nephrin, but also down-regulate of desmin expression.