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Puromycin aminonucleoside

A glomerular epithelial cell toxin

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  • 货号: ajci4584
  • CAS: 58-60-6
  • 别名: 氨基核苷嘌呤霉素; NSC 3056
  • 分子式: C12H18N6O3
  • 分子量: 294.31
  • 纯度: >98%
  • 溶解度: ≥ 14.45 mg/mL in DMSO, ≥ 29.4 mg/mL in EtOH with gentle warming, ≥ 29.5 mg/mL in Water with gentle warming
  • 储存: Store at -20°C
  • 库存: 现货

Background

IC50: N/A


Puromycin aminonucleoside, 3'-Amino-3'-deoxy-N6,N6-dimethyladenosine, is the aminonucleoside portion of the antibiotic puromycin. Puromycin aminonucleoside (PAN)-induced nephrosis in rats can provide a model for investigating the pathogenesis of severe proteinuric conditions.


In vitro: A pervious study used scanning (SEM) and transmission (TEM) electron microscopy to test the in vitro effects of PAN on rat glomerular podocytes. Slices of rat kidney were incubated with PAN. SEM analysis of glomeruli on kidney slices indicated incubation with PAN decreased the number of microvilli on podocyte cell bodies and increased the number of glomeruli. TEM morphometry showed PAN incubation significantly retarded the loss of podocyte foot processes that was observed in control groups [1].


In vivo: In Wistar rats, multiple injections of PAN resulted in sustained severe proteinuria and FSGHS lesions of their glomeruli. In PVG/c rats, a higher PAN dose was needed to induce chronic proteinuria. In acute PAN nephrosis induced by a single intravenous injection of PAN the mesangium of Wistar rats showed large amounts of lipid in contrast to a few small mesangial lipid droplets in nephrotic PVG/c rats. Moreover, after injection of colloidal carbon in nephrotic PVG/c rats no enhanced carbon accumulation was found in the mesangium when compared to nonproteinuric controls [2].


Clinical trial: N/A


IC50:N/A


普鲁霉素氨核苷(Puromycin aminonucleoside, PAN),即普鲁霉素抗生素的氨核苷部分,能引起大鼠肾小球上皮细胞的病变,提供一种研究重度蛋白尿病理机制的模型。


体外研究:一项早期的研究使用扫描(SEM)和透射(TEM)电子显微镜测试了PAN对大鼠肾小球上皮细胞的体外影响。大鼠肾脏切片被PAN孵育。对肾切片上的肾小球的SEM分析表明,PAN孵育减少了上皮细胞体的微绒毛数量,增加了肾小球的数量。TEM形态学显示,与对照组相比,PAN孵育明显减缓了上皮细胞足突丢失的速度[1]。


体内研究:在Wistar大鼠中,多次注射PAN导致持续的严重蛋白尿和肾小球FSGHS病变。在PVG/c大鼠中,需要更高的PAN剂量才能诱导慢性蛋白尿。在通过单次静脉注射PAN诱导急性PAN肾病的Wistar大鼠中,与蛋白尿PVG/c大鼠相比,Wistar大鼠的系膜中显示出大量的脂质,而PVG/c大鼠的系膜中只有少量的小的脂质滴。此外,在蛋白尿PVG/c大鼠中注射胶体炭后,与非蛋白尿对照相比,系膜中没有发现增强的碳积累[2]。


临床试验:N/A

参考文献:
[1] Grond J,Muller EW,van Goor H,Weening JJ,Elema JD.? Differences in puromycin aminonucleoside nephrosis in two rat strains. Kidney Int.1988 Feb;33(2):524-9.
[2] Bertram JF,Messina A,Ryan GB.? In vitro effects of puromycin aminonucleoside on the ultrastructure of rat glomerular podocytes. Cell Tissue Res.1990 May;260(3):555-63.

Protocol

Cell experiment [1]:

Cell lines

Madin-Darby canine kidney (MDCK) cells

Preparation method

The solubility of this compound in DMSO is >14.5mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

48 h

Applications

In vector- and PMAT-transfected MDCK cells, Puromycin aminonucleoside (PAN) exhibited cell cytotoxicity with the IC50 values of 48.9 ± 2.8 and 122.1 ± 14.5 μM, respectively. PAN (250 μM) was toxic to both PMAT-expressing and vector-transfected cells. Puromycin aminonucleoside uptake in PMAT-expressing cells was four fold higher at pH 6.6 than that at pH 7.4.

Animal experiment [2,3]:

Animal models

Nephrosis rats

Dosage form

Intravenous injection, 60 mg/kg, 150 mg/kg

Application

In nephrosis rats, the number of podocytes per glomerulus was 90.7 on Day 4 in PAN (8 mg/100 g, i.v.) treated group. The amount of nephrin per glomerulus in PAN-treated nephrosis rats reduced to 0.46 ± 0.06 fmol and 0.35±0.04 fmol on Day 4 and Day 7. The nephrin amount per podocyte was significantly decreased association with the development of proteinuria in Puromycin aminonucleoside nephrosis rats. Rats given PAN (100 mg/kg, s.c.) gained less weight and their serum creatinine levels were higher than the control rats, indicating Puromycin aminonucleoside impaired renal function.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

参考文献:

[1]. Xia L, Zhou M, Kalhorn T F, et al. Podocyte-specific expression of organic cation transporter PMAT: implication in puromycin aminonucleoside nephrotoxicity. American Journal of Physiology-Renal Physiology, 2009, 296(6): F1307-F1313.


[2]. Kawakami, Hirotaka, et al. Dynamics of absolute amount of nephrin in a single podocyte in puromycin aminonucleoside nephrosis rats calculated by quantitative glomerular proteomics approach with selected reaction monitoring mode. Nephrology Dialysis Transplantation 27.4 (2011): 1324-1330.


[3]. Nosaka, Kazuo, et al. An adenosine deaminase inhibitor prevents puromycin aminonucleoside nephrotoxicity. Free Radical Biology and Medicine 22.4 (1997): 597-605.

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