氯化血根堿作用機(jī)制 - Medchemexpress - MCE中國

1、Product Data SheetSanguinarine chlorideCat. No.: HY-N0052ACAS No.: 5578-73-4分式: CHClNO分量: 367.78作靶點(diǎn): Apoptosis; Autophagy; Bacterial; Parasite作通路: Apoptosis; Autophagy; Anti-infection儲(chǔ)存式: 4C, protect from light* In solvent : -80C, 6 months; -20C, 1 month (protect from light)溶解性數(shù)據(jù)體外實(shí)驗(yàn) H2O : 0.1 mg/mL

2、 (insoluble)DMSO : 5 mg/mL (13.60 mM; Need ultrasonic)SolventMass1 mg 5 mg 10 mgConcentration制備儲(chǔ)備液1 mM 2.7190 mL 13.5951 mL 27.1902 mL5 mM 0.5438 mL 2.7190 mL 5.4380 mL10 mM 0.2719 mL 1.3595 mL 2.7190 mL請根據(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 (p

3、rotect from light)。-80C 儲(chǔ)存時(shí)，請?jiān)?6 個(gè)內(nèi)使，-20C 儲(chǔ)存時(shí)，請?jiān)?1 個(gè)內(nèi)使。體內(nèi)實(shí)驗(yàn)請根據(jù)您的實(shí)驗(yàn)動(dòng)物和給藥式選擇適當(dāng)?shù)娜芙獍?。以下溶解案都請先按?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；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的式助溶1. 請依序添加每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 0

4、.5 mg/mL (1.36 mM); Clear solution此案可獲得 0.5 mg/mL (1.36 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 5.0 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. 請依序添加每種溶劑： 10% DMSO 90% (20% SBE-CD in saline)Solubility: 0.5 mg/mL (1.36 mM); Clear solution此案可獲得 0.5 mg/mL (1.3

5、6 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 5.0 mg/mL 的澄 DMSO 儲(chǔ)備液加到 900 L 20% 的 SBE-CD 理鹽溶液中，混合Page 1 of 2 www.MedChemE均勻。BIOLOGICAL ACTIVITY物活性 Sanguinarine chloride種來 于 Sanguinaria Canadensis 的物堿，可通過激活活性氧 (ROS) 的產(chǎn)來刺激細(xì)胞凋亡。Sanguinarine 誘導(dǎo)的細(xì)胞凋亡與 JNK 和 NF-B 的活化有關(guān)。IC & Target Apoptosis1體外研究 Sanguinarine (SAN

6、G)-induced apoptosis is associated with the activation of JNK and NF-B signal pathways.Todetermine the effects of Sanguinarine on cell viability, 22B-cFluc cells are stimulated with different concentrations ofSanguinarine for 24 h, and then a CKK-8 assay is performed. The treatment with Sanguinarine

7、 decreases theproliferation of 22B cells in a dose-dependent manner. Meanwhile, the cytosolic extracts of 22B-cFluc cells treatedwith different dose of Sanguinarine are measured to detect cellular caspase-3 activity using Ac-DEVD-pNA, which is avalidated caspase-3 substrate. The absorbance at 450 nm

8、 increases in a dose-dependent manner, indicatingincreased caspase-3 activity stimulated by Sanguinarine1.體內(nèi)研究 To evaluate the apoptosis induced by Sanguinarine (SANG) in vivo, 22B-cFluc cells are inoculated subcutaneously intoone flank of nude mice and xenograft models are allowed to establish. Mic

9、e are treated intravenously with 10 mg/kg of Sanguinarine. At 24, 48 and 72 h after treatment, bioluminescent imaging is performed after i.p. injection of micewith 150 mg/kg of D-luciferin substrate. Sanguinarine treatment induces an obvious increase of luminescent signal asearly as 48 h after initi

10、al treatment. A sustained bioluminescent imaging (BLI) intensity increased is observedthroughout the course of experiment. At 72 h after treatment, the tumors are collected and subjected to TUNELstaining for evaluating apoptosis. Compared with the control tumors, the group treated with Sanguinarine

11、exhibitssignificantly more cell apoptosis, indicated by the increased green signals from the sporadic apoptotic cells1.PROTOCOLKinase Assay 1 The caspase-3 activity is measured using a caspase-3 activity assay kit. Briefly, the cells treated by differentconcentrations of Sanguinarine (0.5 M, 1 M, 2

12、M, 4 M) or control DMSO are collected, washed and lysed in alysis buffer for 30 min on ice. The supernatants are then collected by centrifuging at 1,2000 rpm for 10 min. The Ac-DEVD-pNA (2 mM) is added to each sample and incubated at 37C for 1 h. The optical density (OD) of each sample isfinally qua

13、ntified at a wavelength of 405 nm using a spectrophotometer. The p-NA standard is used to calibrate thecaspase-3 activity of each sample1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 The cell viability of Sanguinarine is determined by CC

14、K-8 assay using a cell counting kit-8. Briefly, 22B-cFluc cells areseeded in a 96-well plate (5103 cells/well) and treated with different concentrations of Sanguinarine (0.5 M, 1 M,2 M, 4 M) for 24 h. Then 10 mL CKK-8 is added to each well for 4 h and the absorbance at 450 nm is measuredwith a micro

15、plate reader. The optical density (OD) values are determined to reflect the viable cell populations fromeach well1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice1Administration 1 Xenografted tumor models are prepared by injection of 1107 22

16、B-cFluc cells suspended in PBS into nude mouse(n=6). After tumors reach a volume of approximately 100 mm3, Sanguinarine (10 mg/kg) is i.v. injected into mice.After injection for 24 h, 48 h and 72 h, mice are given a single i.p. dose of 150 mg/kg D-luciferin and bioluminescencePage 2 of 3 www.MedChem

17、Eimaging are performed using a Xenogen Lumina II system. The signal intensity in the region of interest is expressedusing the Living Image software 4.1. For the anti-tumor therapy studies, one group of tumor-bearing mice (n=6)receive intravenously 10 mg/kg of Sanguinarine every other day throughout

18、the experimental period, while thecontrol group of mice (n=6) receive DMSO only. Tumor growth measurement is calculated1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.REFERENCES1. Wang Y, Noninvasive bioluminescence imaging of the dynamics of sanguinarine induced apoptosis via activation of reactive o