三七总皂苷对顺铂肾损害大鼠肾组织差异表达蛋白质的影响

目的 探讨三七总皂苷（panax notoginsenosides,PNS）对顺铂肾损害大鼠肾组织差异表达蛋白质的影响。方法 实验大鼠随机分为正常对照组、顺铂模型组和PNS治疗组,对大鼠给药处理10 d后,检测大鼠血清尿素氮（BUN）、肌酐（Scr）和尿N-乙酰-β-氨基葡萄糖苷酶（NAG）的水平,并做肾脏病理检查;采用SELDI-TOF-MS技术筛选大鼠肾组织的差异表达蛋白质,并通过MALDI-TOF-MS/MS、Western blot实验予以鉴定。结果 顺铂模型组大鼠血清BUN、Scr和尿NAG的水平均显著高于正常对照组（P均〈0.05）。电镜下可见肾小管上皮细胞的线粒体明显损伤,说明顺铂肾损害大鼠模型制作成功。PNS干预可使大鼠血清BUN、Scr和尿NAG的水平显著低于顺铂模型组（P〈0.05）,肾小管上皮细胞的线粒体损害程度较顺铂模型组明显改善,提示PNS对其有保护作用。筛选出顺铂模型组与正常对照组肾组织差异表达的蛋白质20个,其中7个蛋白质在顺铂模型组中的表达下调2倍以上;顺铂模型组与PNS治疗组肾组织差异表达的蛋白质18个,其中11个蛋白质在PNS治疗组中的表达下调;有6个共同的差异表达蛋白质在顺铂模型组较正常对照组的表达上调或下调,在PNS治疗组可回调到接近正常对照组水平。差异表达蛋白质m/z 10815.42被鉴定为线粒体热休克蛋白,m/z 16021.67被鉴定为血红蛋白β1亚基、血红蛋白β2亚基。结论 顺铂肾损害可伴随多种蛋白质的表达变化,这些差异表达蛋白质可能与顺铂损害肾脏以及PNS的保护作用有关。通过对其分离和鉴定,进一步了解其性质,将有助于全面、系统地探讨顺铂肾损害以及PNS保护作用的机制。

Effect of Panax Notoginsenosides on renal proteins differentially expressed in rats experiencing cisplatin-induced nephrotoxicity

Objective To understand the effects of Panax Notoginsenosides （PNS） on renal proteins differentially expressed in rats experiencing cisplatin-induced nephrotoxicity. Methods Rats were randomly divided into three groups and treated with saline,cisplatin alone or cisplatin with PNS. At 10 days after treatment,renal tissue was examined for pathology,and levels of serum BUN,Scr,and urine NAG were determined. Renal proteins for which expression levels changed in response to cisplatin treatment were screened using SELDI-TOF-MS and identified using MALDI-TOF-MS/MS and Western blotting. Results We successfully created a rat model of cisplatin-induced nephrotoxicity, and PNS reduced the drug-induced damage. Mass spectrometry identified 20 renal proteins differentially expressed between the saline and cisplatin groups, as well as 18 proteins differentially expressed between the cisplatin and cisplatin＋ PNS groups. Of these proteins, 6 were up-or down-regulated by cisplatin treatment, and PNS returned their expression close to the levels in the saline group. One of these proteins（rrdz 10815.42） was identified as mitochondrial heat shock protein, while another （m/z 16021.67） was identified as hemoglobin subunit beta-1 and beta-2. Conclusions These differentially expressed renal proteins may mediate cisplatin-induced nephrotoxicity as well as the protective effects of PNS. Further studies should verify these leads and examine the cellular pathways involved.