氯化血根碱作用机制 - Medchemexpress - MCE中国

1、Product Data SheetSanguinarine chlorideCat. No.: HY-N0052ACAS No.: 5578-73-4分式: CHClNO分量: 367.78作靶点: Apoptosis; Autophagy; Bacterial; Parasite作通路: Apoptosis; Autophagy; Anti-infection储存式: 4C, protect from light* In solvent : -80C, 6 months; -20C, 1 month (protect from light)溶解性数据体外实验 H2O : 0.1 mg/mL

2、 (insoluble)DMSO : 5 mg/mL (13.60 mM; Need ultrasonic)SolventMass1 mg 5 mg 10 mgConcentration制备储备液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 (p

3、rotect from light)。-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: 0

4、.5 mg/mL (1.36 mM); Clear solution此案可获得 0.5 mg/mL (1.36 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 5.0 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: 0.5 mg/mL (1.36 mM); Clear solution此案可获得 0.5 mg/mL (1.3

5、6 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 5.0 mg/mL 的澄 DMSO 储备液加到 900 L 20% 的 SBE-CD 理盐溶液中，混合Page 1 of 2 www.MedChemE均匀。BIOLOGICAL ACTIVITY物活性 Sanguinarine chloride种来 于 Sanguinaria Canadensis 的物碱，可通过激活活性氧 (ROS) 的产来刺激细胞凋亡。Sanguinarine 诱导的细胞凋亡与 JNK 和 NF-B 的活化有关。IC & Target Apoptosis1体外研究 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.体内研究 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