天津医药
天津醫藥
천진의약
TIANJIN MEDICAL JOURNAL
2014年
6期
526-529
,共4页
艾辰%谭小月%张勉之%张大宁
艾辰%譚小月%張勉之%張大寧
애신%담소월%장면지%장대저
五味子科%足细胞%膜蛋白质类%阿霉素%nephrin%desmin
五味子科%足細胞%膜蛋白質類%阿黴素%nephrin%desmin
오미자과%족세포%막단백질류%아매소%nephrin%desmin
schisandraceae%podocytes%membrane proteins%adriamycin%nephrin%desmin
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/L ADR）、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。
目的：觀察五味子醇提液（SE）對阿黴素（ADR）誘導的體外足細胞損傷中足細胞裂孔膜蛋白nephrin和骨架蛋白desmin錶達的影響，併探討其作用機製。方法體外ADR作用于足細胞24 h，造成足細胞損傷後，加入含SE的培養基，繼培養24 h。設對照組（0 mg/L ADR）、模型組（0.25 mg/L ADR）、SE榦預組（250 mg/L SE+0.25 mg/L ADR）、SE組（250 mg/L SE）。免疫熒光法測定各組nephrin的錶達；Western Blot檢測各組nephrin和desmin蛋白錶達；RT-PCR檢測各組nephrin和desmin mRNA錶達。結果免疫熒光結果顯示對照組和SE組nephrin呈顆粒狀或線性分佈于細胞膜，模型組較對照組、SE組和SE榦預組錶達彊度明顯減少。模型組nephrin蛋白和mRNA錶達較對照組顯著減少，SE榦預組nephrin蛋白和mRNA錶達水平較模型組增加。模型組desmin蛋白和mRNA錶達較對照組顯著增加，SE榦預組desmin蛋白和mRNA錶達水平較模型組顯著減少（均P<0.05）。SE和SE榦預組的nephrin和desmin蛋白和mRNA錶達水平與對照組差異無統計學意義。結論250 mg/L SE對體外培養足細胞無損傷作用，且SE能拮抗ADR對足細胞的損傷，這可能與SE可以上調nephrin和下調desmin的錶達有關。
목적：관찰오미자순제액（SE）대아매소（ADR）유도적체외족세포손상중족세포렬공막단백nephrin화골가단백desmin표체적영향，병탐토기작용궤제。방법체외ADR작용우족세포24 h，조성족세포손상후，가입함SE적배양기，계배양24 h。설대조조（0 mg/L ADR）、모형조（0.25 mg/L ADR）、SE간예조（250 mg/L SE+0.25 mg/L ADR）、SE조（250 mg/L SE）。면역형광법측정각조nephrin적표체；Western Blot검측각조nephrin화desmin단백표체；RT-PCR검측각조nephrin화desmin mRNA표체。결과면역형광결과현시대조조화SE조nephrin정과립상혹선성분포우세포막，모형조교대조조、SE조화SE간예조표체강도명현감소。모형조nephrin단백화mRNA표체교대조조현저감소，SE간예조nephrin단백화mRNA표체수평교모형조증가。모형조desmin단백화mRNA표체교대조조현저증가，SE간예조desmin단백화mRNA표체수평교모형조현저감소（균P<0.05）。SE화SE간예조적nephrin화desmin단백화mRNA표체수평여대조조차이무통계학의의。결론250 mg/L SE대체외배양족세포무손상작용，차SE능길항ADR대족세포적손상，저가능여SE가이상조nephrin화하조desmin적표체유관。
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.
