毒毛旋花苷G作用机制 - Medchemexpress - MCE中国

1、Product Data SheetOuabain OctahydrateCat. No.: HY-B0542CAS No.: 11018-89-6分式: CHO分量: 728.77作靶点: Na+/K+ ATPase; Autophagy作通路: Membrane Transporter/Ion Channel; Autophagy储存式: Powder -20C 3 years4C 2 yearsIn solvent -80C 6 months-20C 1 month溶解性数据体外实验 DMSO : 35 mg/mL (48.03 mM)* means soluble, but satur

2、ation unknown.SolventMass1 mg 5 mg 10 mgConcentration制备储备液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请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；旦配成溶液，请分装保存，避免反复冻融造成的产品失效。储备液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 储存时，请在 6 个内使，-20C 储存时，请在 1 个内使。体内实验请根据

3、您的实验动物和给药式选择适当的溶解案。以下溶解案都请先按照 In Vitro 式配制澄清的储备液，再依次添加助溶剂：为保证实验结果的可靠性，澄 的储备液可以根据储存条件，适当保存；体内实验的作液，建议您现现配，当天使； 以下溶剂前显的百分 指该溶剂在您配制终溶液中的体积占；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的式助溶1. 请依序添加每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (3.43 mM); Clear solution此案可获得 2.5 mg/mL (3.43 mM，饱和度

4、未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 400 L PEG300 中，混合均匀；向上述体系中加50 L Tween-80，混合均匀；然后继续加 450 L 理盐定容 1 mL。2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (3.43 mM); Clear solutionPage 1 of 2 www.MedChemE此案可获得 2.5 mg/mL (3.43 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L

5、 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 20% 的 SBE-CD 理盐溶液中，混合均匀。3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (3.43 mM); Clear solution此案可获得 2.5 mg/mL (3.43 mM，饱和度未知) 的澄 溶液，此案不适于实验周 期在半个以上的实验。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 油中，混合均匀。BIOLOGICAL ACTIVITY物活性 Ouabain OctahydrateNa+/

6、K+-ATPase 抑制剂，常于治疗充性衰竭。体外研究 Ouabain (100 M) induces NLRP3 inflammasome activation and IL-1 release in macrophages. Ouabain-inducedNLRP3 inflammasome activation is mediated through K+ efflux1. Ouabain (3 nM) alters the expression of EMTmarkers in NHK and ADPKD cells, and modifies cell-cell adhesion p

7、roperties in ADPKD. Moreover, ouabain enhancesmigration of ADPKD cells, selectively modulates tight junctions, and modulates adherens junctions in ADPKD cells in aselective manner. Ouabain also activates TGF-Smad3 signaling, alters TER in ADPKD cells2. Ouabain (25, 50 or 100nM) treatment significant

8、ly reduces cell proliferation and viability in Raji cells in a dose-dependent manner, with IC50of 76.484.03 nM. Ouabain increases the number of apoptotic cells, induces autophagy, and upregulates Beclin-1 inRaji cells4.体内研究 Ouabain (3 mg/kg) significantly decreases cardiac contractile force with an

9、enlarged LVESD when mice are primed with LPS. IL-1 deficiency attenuates ouabain-induced cardiac dysfunction and injury. IL-1 secreted by infiltrated macrophages contributes to ouabain-induced cardiac inflammation. Deficiency of NLRP3 and Casp1 attenuatesouabain-induced cardiac dysfunction and macro

10、phage infiltration1. Ouabain (30 g/kg, i.p.) modulates ABCB1activity in thymocytes of Wistar rats and it has the same effect on Swiss mice at 300 g/kg. After 14 days of ouabaintreatment, the MAP of rats is significantly elevated3.PROTOCOLCell Assay 4 Cell viability is determined using a Cell Countin

11、g Kit-8 assay. Briefly, 100 L Raji cells (5104/mL) are seeded intriplicate in a 96-well plate 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 is added to each well, and thec

12、ells are incubated for an additional 3 h at 37C. Optical density (OD) values at 450 nm are subsequently measured,and each ouabain concentration is assessed in triplicate. Raji cells cultured in medium without drug served ascontrols. Cell viability is calculated according to the following formula: In

13、hibition rate (%)=1 (OD450(sample) OD450(blank)/(OD450(control) OD450(blank) 100.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice3Administration 3 A total of 8 mice are used in the present study, 4 in the control group and 4 in the ouabain-tr

14、eated group. Animalsare maintained under standard laboratory conditions, with room temperature controlled (22C), and subjected to 12h light-dark cycles with ad libitum access to food and water. At 24 h subsequent to the intraperitoneal injection with300 g/kg of ouabain or PBS, the Swiss mice are sac

15、rificed by barbiturate overdose (86 mg/kg intraperitonealinjection of pentobarbital). The mesenteric lymph nodes and thymi are immediately removed and softly dissociated.The remaining cells are washed in PBS and centrifuged at 200 g. The pellet is suspended in ice-cold RPMI-1640culture medium supple

16、mented with 10% FBS until required for the activity assays.Page 2 of 3 www.MedChemERat3Male 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 in ouab

17、ain and controlgroups) and 8 for chronic treatment (n=4 rats/group in ouabain and control groups). Animals are maintained understandard laboratory conditions, with room temperature controlled (22C), and subjected to 12 h light-dark cycles withad libitum access to food and water. Prior to the first i

18、njection at 24 h and 7 and 14 days subsequent to the injection,the rats have their blood pressure measured by a computerized tail-cuff method. The animals are sacrificed bybarbiturate overdose (86 mg/kg intraperitoneal injection of pentobarbital) after 24 h (acute treatment) or 14 days(chronic treat

19、ment) of ouabain injections, and the mesenteric lymph nodes, thymi and blood are collected. Fullexcisions of thymi and partial excisions of mesentheric lymph nodes are performed, while blood samples arecollected by caudal venous puncture prior to animals sacrifice.MCE has not independently confirmed

20、 the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Med Sci Monit. 2019 Dec 11;25:9426-9434.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Kobayashi M, et al. The cardiac glycoside ouabain activates NLRP3 inflammasomes and promotes cardiac inflammation and dysfunction. PLoS One. 2017May 11;12(5):e0176676.2. Venugopal J, et al. Ouabain promotes partial e