生物活性

TEMPOL is a superoxide dismutase mimetic that belongs to a class of non-thiol-containing radiation protectors, and has the ability to permeate the membrane. Superoxide scavenger that displays neuroprotective, anti-inflammatory and analgesic effects. In the presence of TEMPOL, cerebral vessel remodeling is blocked and collagen IV mRNA expression levels are elevated in normotensive and hypertensive rat studies. TEMPOL has been shown to restore the oxidative stress and cardiac dysfunction induced by TNF-α and in PC-12 cells has revealed neuroprotective effects. Tempol is a low molecular weight, watersoluble, membrane-permeable, and metal-independent superoxide dismutase mimeticthat has shown antioxidant effects in a variety of disease models involvingoxidative stress.

Antiproliferativeeffect of Tempol[1]

体外研究

体内研究

Saline

激酶实验

细胞实验

Cellculture[2]

The H9c2 cells were cultured in Dulbecco’smodified Eagle’s medium (DMEM) containing 10% fetal bovine serum, 100units/mlpenicillin, and 100units/ml streptomycin. Cultures were grown at 37 °C in ahumidified atmosphere incubator with 95% air and 5% CO₂. The medium waschanged every 2–3 days.

Cellviability assay

H9c2 cells were seeded in 96-well plates ata density of 1.5×10⁵ cells/well. After incubation for 24 h at 37°Cin a CO₂ incubator, H9c2 cells were incubated with differentconcentrations of Tempol (0.1, 0.33, 1, 3.3, and 10μmol/L) for 1 h except forthe control cells receiving medium instead. Then, H9c2 cells were placed in ahypoxia incubator with 95% N2 and 5% CO₂ for 24 h. Control cellswere cultured for 24 h in normoxic condition. After that, the cells were washedwith cold PBS, and then incubated in 100μL with MTT solution (0.5mg/ml inmedium) for 4 h. Finally, the violet crystals were dissolved with 100μl DMSO.The absorbance was measured at 570nm with a microplate reader. Cell viabilityis expressed as the percentage of control.

动物实验

Animals[3]

Forty female Wistar albino rats aged 12weeks, weighing 250-300g, were caged individually under controlledenvironmental conditions with 20-22 °C temperature and humidity and with a 12-hlight/dark cycle and were fed ad libitum. The rats were separated randomly intofive groups with eight rats each: Group I: sham, Group II: ischemia (I), GroupIII: ischemia/reperfusion (I/R), Group IV: I/R+ Tempol 30 mg/kg, Group V: I/R +Tempol 50 mg/kg.

Experimentalprotocol

Each rat was weighed and anesthetized withintramuscular ketamine hydrochloride (50 mg/kg) and xylazine hydrochloride (10mg/kg). The rats were placed in the dorsal recumbent position. A 1.5 cm midlinelongitudinal laparotomy incision was made under sterile conditions. The abdomenwas opened and the large intestines were separated attentively and removed outof abdomen. The uterine horns and adnexa were observed for 1 minute in the Shamgroup and then closed with 3-0 surgical suture. In the experimental groups(Group I-V), vascular clamps were placed just beneath the ovaries and over theuterine horns to provide total ischemia. Incisions were closed with 4/0 silksuture.

In group II (Ischemia group), ischemia wasinduced for 3 hours using a torsion model stimulated by the application of atraumatic vascular clamp to the vascular pedicle 1 cm above and below theovary. The incision was closed with 4/0 silk suture. In group III (I/R group), ischemiawas performed for 3 hours, followed by reperfusion for 3 hours. In group IV(I/R and Tempol 30 mg/kg i.p. group), ischemia was induced for 3 hours as inGroup II, followed by the administration of Tempol 30 mg/kg i.p. 30 min priorto 3 hours of reperfusion. In group V (I/R and Tempol 50 mg/kg i.p. group),Tempol 50 mg/kg i.p. was administered 30min prior to 3 hours of reperfusion.

After surgery, all the rats weresacrificed. Blood samples were obtained through cardiac puncture andcentrifuged at 1,500 x g for 10 minutes. Supernatants were portioned and storedat -80 °C. Both ovaries were extirpated. One of the ovaries from each rat werestored in a freezer at -80 °C for biochemical analysis and the other was fixedin formaldehyde solution 10% for histological examination.

Biochemicalanalysis

The ovarian tissues were weighed andhomogenized in ice-cold phosphate-buffered saline at pH 7.4 (10% w/v) andcentrifuged at 1,500 x g for 15 min at 4°C. Supernatants were removed foranalysis. Protein content of supernatants was determined using bovine serumalbumin as standard.

SerumTOS, TAS and OSI levels

Serum total oxidant status (TOS) and totalantioxidant status (TAS) were measured by using a novel automated colorimetricmethod. The TOS results were expressed as micromolar hydrogen peroxideequivalent per liter (μmol H₂O₂ Eq/l) and the TAS levelswere expressed as mmol Trolox Eq/l. The ratio of TOS to TAS was accepted as theoxidative stress index (OSI) and was calculated as follows: OSI (arbitraryunit) =TOS (μmol H₂O₂ Eq/l) / TAS (μmol Trolox Eq/l) x100.

不同实验动物依据体表面积的等效剂量转换表（数据来源于FDA指南）

例如，已知某工具药用于小鼠的剂量为88 mg/kg , 则用于大鼠的剂量换算方法：将88 mg/kg 乘以小鼠的Km系数（3），再除以大鼠的Km系数（6），得到该药物用于大鼠的等效剂量44 mg/kg。

参考文献

[1] Gariboldi MB RV, Supino R, Favini E, Monti E. The nitroxide tempol induces oxidative stress, p21(WAF1/CIP1), and cell death in HL60 cells. Free Radic Biol Med. 2000;29(7):633-641.
[more]

体内溶解度
约30 mg/mL

Clinical Trial Information ( data from http://clinicaltrials.gov )

注：以上所有数据均来自公开文献，并不保证对所有实验均有效，数据仅供参考。

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