毒毛旋花苷G作用機(jī)制 - Medchemexpress - MCE中國(guó)

1、Product Data SheetOuabain OctahydrateCat. No.: HY-B0542CAS No.: 11018-89-6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 solvent -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 satur

2、ation unknown.SolventMass1 mg 5 mg 10 mgConcentration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；旦配成溶液，請(qǐng)分裝保存，避免反復(fù)凍融造成的產(chǎn)品失效。儲(chǔ)備液的保存式和期限：-80C, 6 months; -20C, 1 month。-80C 儲(chǔ)存時(shí)，請(qǐng)?jiān)?6 個(gè)內(nèi)使，-20C 儲(chǔ)存時(shí)，請(qǐng)?jiān)?1 個(gè)內(nèi)使。體內(nèi)實(shí)驗(yàn)請(qǐng)根據(jù)

3、您的實(shí)驗(yàn)動(dòng)物和給藥式選擇適當(dāng)?shù)娜芙獍?。以下溶解案都?qǐng)先按照 In Vitro 式配制澄清的儲(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，澄 的儲(chǔ)備液可以根據(jù)儲(chǔ)存條件，適當(dāng)保存；體內(nèi)實(shí)驗(yàn)的作液，建議您現(xiàn)現(xiàn)配，當(dāng)天使； 以下溶劑前顯的百分 指該溶劑在您配制終溶液中的體積占；如在配制過(guò)程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過(guò)加熱和/或超聲的式助溶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 solution此案可獲得 2.5 mg/mL (3.43 mM，飽和度

4、未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲(chǔ)備液加到 400 L PEG300 中，混合均勻；向上述體系中加50 L Tween-80，混合均勻；然后繼續(xù)加 450 L 理鹽定容 1 mL。2. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (3.43 mM); Clear solutionPage 1 of 2 www.MedChemE此案可獲得 2.5 mg/mL (3.43 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L

5、 25.0 mg/mL 的澄 DMSO 儲(chǔ)備液加到 900 L 20% 的 SBE-CD 理鹽溶液中，混合均勻。3. 請(qǐng)依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (3.43 mM); Clear solution此案可獲得 2.5 mg/mL (3.43 mM，飽和度未知) 的澄 溶液，此案不適于實(shí)驗(yàn)周 期在半個(gè)以上的實(shí)驗(yàn)。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲(chǔ)備液加到 900 L 油中，混合均勻。BIOLOGICAL ACTIVITY物活性 Ouabain OctahydrateNa+/

6、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 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.體內(nèi)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.戶使本產(chǎn)品發(fā)表的科研獻(xiàn) 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