1、Hotline: 400-820-3792Inhibitors Agonists Screening Librarieswww.MedChemEOuabain OctahydrateCat. No.: HY-B0542CAS No.: 11018-89-6Synonyms: Acocantherine; G-Strophanthin分式: CHO分量: 728.77作靶點(diǎn): Na+/K+ ATPase; Autophagy作通路: Membrane Transporter/Ion Channel; Autophagy儲(chǔ)存式: Powder -20C 3 years4C 2 yearsIn so

2、lvent -80C 6 months-20C 1 month溶解性數(shù)據(jù)體外實(shí)驗(yàn) DMSO : 35 mg/mL (48.03 mM)* means soluble, but saturation unknown.Mass Solvent1 mg 5 mg 10 mg Concentration制備儲(chǔ)備液1 mM 1.3722 mL 6.8609 mL 13.7218 mL5 mM 0.2744 mL 1.3722 mL 2.7443 mL10 mM 0.1372 mL 0.6861 mL 1.3722 mL請(qǐng)根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲(chǔ)備液，并請(qǐng)注意儲(chǔ)備液的保存式和期限

3、。體內(nèi)實(shí)驗(yàn)請(qǐng)根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥式選擇適當(dāng)?shù)娜芙獍?，配制前?qǐng)先配制澄清的儲(chǔ)備液，再依次添加助溶劑(為保證實(shí)驗(yàn)結(jié)果的可靠性，體內(nèi)實(shí)驗(yàn)的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使；澄清的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)保存；以下溶劑前的百分 指該溶劑在您配制終溶液中的體積占)：1. 請(qǐng)依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (3.43 mM); Clear solution2. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/m

4、L (3.43 mM); Clear solution1/3 Master of Small Molecules 您邊的抑制劑師www.MedChemE3. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (3.43 mM); Clear solutionBIOLOGICAL ACTIVITY物活性 Ouabain OctahydrateNa+/K+-ATPase 抑制劑，常于治療充性衰竭。體外研究 Ouabain (100 M) induces NLRP3 inflammasome activation and IL-1 relea

5、se in macrophages. Ouabain-induced NLRP3 inflammasome activation is mediated through K+ efflux 1. Ouabain (3 nM) alters theexpression of EMT markers in NHK and ADPKD cells, and modifies cell-cell adhesion properties in ADPKD.Moreover, ouabain enhances migration of ADPKD cells, selectively modulates

6、tight junctions, and modulatesadherens junctions in ADPKD cells in a selective manner. Ouabain also activates TGF-Smad3 signaling,alters TER in ADPKD cells 2. Ouabain (25, 50 or 100 nM) treatment significantly reduces cell proliferationand viability in Raji cells in a dose-dependent manner, with IC5

7、0 of 76.484.03 nM. Ouabain increases thenumber of apoptotic cells, induces autophagy, and upregulates Beclin-1 in Raji cells 4.體內(nèi)研究 Ouabain (3 mg/kg) significantly decreases cardiac contractile force with an enlarged LVESD when mice areprimed with LPS. IL-1 deficiency attenuates ouabain-induced card

8、iac dysfunction and injury. IL-1 secretedby infiltrated macrophages contributes to ouabain-induced cardiac inflammation. Deficiency of NLRP3 andCasp1 attenuates ouabain-induced cardiac dysfunction and macrophage infiltration 1. Ouabain (30 g/kg,i.p.) modulates ABCB1 activity in thymocytes of Wistar

9、rats and it has the same effect on Swiss mice at 300g/kg. After 14 days of ouabain treatment, the MAP of rats is significantly elevated 3.PROTOCOLCell Assay 4 Cell viability is determined using a Cell Counting Kit-8 assay. Briefly, 100 L Raji cells (5104/mL) areseeded in triplicate in a 96-well plat

10、e and treated with various concentrations of ouabain (400, 200, 100, 50,25, 12.5, 6.25, 3.13, 1.56, 0.78 and 0.39 nM) for 48 h. Following the 48-h treatment, 10 L CCK-8 reagent isadded to each well, and the cells are incubated for an additional 3 h at 37C. Optical density (OD) values at450 nm are su

11、bsequently measured, and each ouabain concentration is assessed in triplicate. Raji cellscultured in medium without drug served as controls. Cell viability is calculated according to the followingformula: Inhibition rate (%)=1 (OD450(sample) OD450(blank)/(OD450(control) OD450(blank) 100.MCE has not

12、independently confirmed the accuracy of these methods. They are for reference only.Animal Mice 3Administration 3 A total of 8 mice are used in the present study, 4 in the control group and 4 in the ouabain-treated group.Animals are maintained under standard laboratory conditions, with room temperatu

13、re controlled (22C), andsubjected to 12 h light-dark cycles with ad libitum access to food and water. At 24 h subsequent to theintraperitoneal injection with 300 g/kg of ouabain or PBS, the Swiss mice are sacrificed by barbiturateoverdose (86 mg/kg intraperitoneal injection of pentobarbital). The me

14、senteric lymph nodes and thymi areimmediately removed and softly dissociated. The remaining cells are washed in PBS and centrifuged at 200 g. The pellet is suspended in ice-cold RPMI-1640 culture medium supplemented with 10% FBS until2/3 Master of Small Molecules 您邊的抑制劑師www.MedChemErequired for the

15、activity assays.Rat 3Male Wistar rats are treated with daily intraperitoneal injections of 30 g/kg of ouabain or its vehicle,phosphate-buffered saline (PBS). A total of 20 rats are used, 12 for acute treatment (n=6 rats/group inouabain and control groups) and 8 for chronic treatment (n=4 rats/group

16、in ouabain and control groups).Animals are maintained under standard laboratory conditions, with room temperature controlled (22C), andsubjected to 12 h light-dark cycles with ad libitum access to food and water. Prior to the first injection at 24 hand 7 and 14 days subsequent to the injection, the

17、rats have their blood pressure measured by acomputerized tail-cuff method. The animals are sacrificed by barbiturate overdose (86 mg/kg intraperitonealinjection of pentobarbital) after 24 h (acute treatment) or 14 days (chronic treatment) of ouabain injections,and the mesenteric lymph nodes, thymi a

18、nd blood are collected. Full excisions of thymi and partial excisionsof mesentheric lymph nodes are performed, while blood samples are collected by caudal venous punctureprior to animals sacrifice.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Kobayashi M, et al. The cardiac glycoside ouabain activates NLRP3 inflammasomes and promotes cardiac inflammation anddysfunction. PLoS One. 2017 May 11;12(5):e0176676.2. Venugopal J, et al. Ouabain promotes partial epithelial to mesenchymal tran