異葒草素作用機(jī)制 - Medchemexpress - MCE中國(guó)

1、Product Data SheetIsoorientinCat. No.: HY-N0767CAS No.: 4261-42-1分式: CHO分量: 448.38作靶點(diǎn): COX作通路: Immunology/Inflammation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 : 100 mg/mL (223.03 mM)* means soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentr

2、ation制備儲(chǔ)備液1 mM 2.2303 mL 11.1513 mL 22.3025 mL5 mM 0.4461 mL 2.2303 mL 4.4605 mL10 mM 0.2230 mL 1.1151 mL 2.2303 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)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

3、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 (5.58 mM); Clear solution此案可獲得 2.5 mg/mL (5.58 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMS

4、O 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 (5.58 mM); Clear solutionPage 1 of 2 www.MedChemE此案可獲得 2.5 mg/mL (5.58 mM，飽和度未知) 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 900 L 20% 的 SBE-CD 理鹽

5、溶液中，混合均勻。BIOLOGICAL ACTIVITY物活性 Isoorientin種有效的 COX-2 抑制劑，IC50 值為 39 M。IC & Target COX-239 M (IC50)體外研究 Isoorientin is a Selective Inhibitor of Cyclooxygenase-2 (COX-2) from the Tubers of Pueraria tuberosa1. PANC-1 andPATU-8988 cells are grown for 24 hours in the presence of Isoorientin (0, 20, 40,

6、80, and 160 M), and a CCK8solution is added. The cell viability decreases significantly at the concentrations of 20, 40, 80, and 160 M. After thecells are cultured with Isoorientin (0, 20, 40, 80, and 160 M for PANC-1; 0, 20, 40, 80, 160, and 320 M for PATU-8988) for 24 hours, the expression of p-AM

7、PK and AMPK is assessed by Western blotting. After the Isoorientintreatment, the p-AMPK expression is increased. Then, in the shRNA group, the concentration of 80 M is used todetect the effects of Isoorientin. The expression levels of AMPK and p-AMPK are much lower in the shRNA group thanin the wild

8、-type PC cells (WT) and the group that is transfected with a negative control lentivirus (NC)2.體內(nèi)研究 Animals treated with Isoorientin at 10 mg/kg and 20 mg/kg body weight have a statistically significant reduction inpaw edema, with a mean peak thickness of 1.190.05 mm and 1.080.04 mm, respectively. T

9、his indicated thatIsoorientin significantly attenuates paw edema compared with the control group3.PROTOCOLCell Assay 2 PANC-1 and PATU-8988 cells are plated onto 96-well plates. Each well contain 5,000 cells and 200 L of themedium with 10% FBS. When the cells of each well reach 70% confluency, the m

10、edium is changed, and FBS-freemedium with different concentrations of Isoorientin is added. After 24 hours, the cells are washed with PBS once, themedium containing Isoorientin is discarded, and 100 L of FBS-free medium with 10 L of the Cell Counting Kit 8(CCK8) reagent are added. The cells are incu

11、bated for another 1-2 hours at 37C, and the absorbance of each well isdetected using an ELISA reader at 490 nm. Cell viability is expressed as the fold change of absorbance2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice3Administration 3 In

12、 the case of paw edema model the Isoorientin or Celecoxib is given intraperitoneally and Carrageenan is injectedinto the paw directly one hour later. In air pouch model all the treatments are given along with Carrageenan directlyinto the pouch cavity. Isoorientin is injected three hours earlier than

13、 injection of Carrageenan into the pouch cavity.Isoorientin and Celecoxib are administered into the mice air pouch. The stock solutions of Isoorientin (100 mg/mL)and Celecoxib (100 mg/mL) are prepared in DMSO and further dilutions are made at the time of treatments. Animalsare divided, into 5 differ

14、ent groups as follows: control (DMSO treated); Carrageenan (0.5 mL of 1.5% (w/v)Carrageenan in saline) treated; Carrageenan+Celecoxib (20 mg/kg body weight) treated; Carrageenan+Isoorientin (10mg/Kg body weight) treated; Carrageenan+Isoorientin (20 mg/Kg body weight) treated.MCE has not independentl

15、y confirmed the accuracy of these methods. They are for reference only.REFERENCESPage 2 of 3 www.MedChemE1. Sumalatha M, et al. Isoorientin, a Selective Inhibitor of Cyclooxygenase-2 (COX-2) from the Tubers of Pueraria tuberosa. Nat Prod Commun. 2015Oct;10(10):1703-4.2. Ye T, et al. Isoorientin indu

16、ces apoptosis, decreases invasiveness, and downregulates VEGF secretion by activating AMPK signaling in pancreatic cancercells. Onco Targets Ther. 2016 Dec 12;9:7481-7492.3. Anilkumar K, et al. Evaluation of Anti-Inflammatory Properties of Isoorientin Isolated from Tubers of Pueraria tuberosa. Oxid Med Cell Longe