穿心莲内酯作用机制 - Medchemexpress - MCE中国

1、Product Data SheetAndrographolideCat. No.: HY-N0191CAS No.: 5508-58-7分式: CHO分量: 350.45作靶点: NF-B; SARS-CoV; Influenza Virus; Autophagy作通路: NF-B; Anti-infection; 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 : 50 mg/mL (142.67 mM; Need ultrasonic)H2O : 0

2、.1 mg/mL (insoluble)SolventMass1 mg 5 mg 10 mgConcentration制备储备液1 mM 2.8535 mL 14.2674 mL 28.5347 mL5 mM 0.5707 mL 2.8535 mL 5.7069 mL10 mM 0.2853 mL 1.4267 mL 2.8535 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。

3、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: 2.5 mg/mL (7.13 mM); Clear solution此案可获得 2.5 mg/mL (7.13

4、 mM，饱和度未知) 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 (7.13 mM); Clear solutionPage 1 of 2 www.MedChemE此案可获得 2.5 mg/mL (7.13 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，

5、取 100 L 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 (7.13 mM); Clear solution此案可获得 2.5 mg/mL (7.13 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 Andrographolide种

6、 NF-B 抑制剂，通过共价修饰内细胞中 p50 的半胱氨酸残 抑制 NF-B 活化，不影 响IB 降解或 p50/p65 核易位。Andrographolide 具有抗病毒作。IC & Target p50体外研究 Andrographolide (AP) concentration-dependently suppresses receptor activator of nuclear factor kappa B ligand(RANKL)-mediated osteoclast differentiation and bone resorption in vitro and reduc

7、es the expression of osteoclast-specific markers. Andrographolide attenuates inflammation by inhibition of TNF-induced NF-B activation throughcovalent modification of reduced Cys62 of p50, without affecting IB degradation or p50/p65 nuclear translocation.Andrographolide also inhibits the ERK/MAPK si

8、gnalling pathway without affecting p38 or JNK signalling.Andrographolide inhibits osteoclast differentiation of RAW 264.7 cells in a concentration-dependent manner.Andrographolide suppresses osteoclast formation in a concentration-dependent manner without any obviouscytotoxic effects, in both BMMs a

9、nd RAW 264.7 cells. Andrographolide treatment substantially reduces the area ofbone resorption. Only approximately 30% of the bone resorption observed in the control group is achieved aftertreatment with 2.5 M Andrographolide. Osteoclastic bone resorption is almost completely inhibited after treatme

10、ntwith 10 M Andrographolide1.体内研究 Treatment with Andrographolide (5 or 30 mg/kg) reduces the extent of bone loss induced by LPS. Moreover,Andrographolide slightly increases the BMD and cortex thickness compared to LPS treatment. Histologicalexamination confirms the protective effects of Andrographol

11、ide on LPS-induced bone loss. LPS injection leads toinflammatory bone erosion and increased numbers of TRAP-positive osteoclasts1.PROTOCOLKinase Assay 1 In vitro osteoclastogenesis assays are preformed to examine the effects of Andrographolide on osteoclastdifferentiation. Bone marrow macrophages (B

12、MM) cells are prepared. Briefly, cells extracted from the femur andtibiae of a 6-week-old C57/BL6 mouse are incubated in complete cell culture media and 30 ng/mL M-CSF in a T-75cm2 flask for proliferation. When changing the medium, the cells are washed in order to deplete residual stromalcells. Afte

13、r reaching 90% confluence, cells are washed with PBS three times and trypsinized for 30 min to harvestBMMs. Cells adhering to the bottom of the dish are classified as BMMs; these BMMs are plated in 96-well plates at adensity of 8103 cells per well in triplicate and incubated in a humidified incubato

14、r containing 5% CO2 at 37C for 24h. The cells are then treated with various concentrations of Andrographolide (0, 2.5, 5, or 10 M) plus M-CSF (30ng/mL) and RANKL (50 ng/mL). After 5 days, cells are fixed and stained for tartrate-resistant acid phosphatase (TRAP)activity. TRAP-positive multinucleated

15、 cells with more than five nuclei are counted as osteoclasts1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Cell Assay 1 Effects of Andrographolide on cell proliferation are determined with a CCK-8. BMMs are plated in 96-well plates at aPage 2 of 3 ww

16、w.MedChemEdensity of 3103 cells per well in triplicate. Twenty-four hours later, the cells are treated with increasingconcentrations of Andrographolide (0, 2.5, 5, 10 or 20 M) for 2 days. Next, 10 L CCK-8 is added to each well, andthe plates are then incubated at 37C for an additional 2 h. The optic

17、al density (OD) is then measured with an ELX800absorbance microplate reader at a wavelength of 450 nm (650 nm reference). The cell viability is calculated1.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Mice1Administration 1 C57BL/6 mice (8 week

18、s old) are divided into four groups of seven mice each. Mice are injected i.p. withAndrographolide (5 or 30 mg/kg body weight) or PBS as a control 1 day before injection of LPS (5 g/g bodyweight). Andrographolide or PBS is injected intraperitoneally every other day for 8 days. LPS is injectedintrape

19、ritoneally on days one and four. All mice are killed 8 days after the initial LPS injection, and the left femurs ofall animals are scanned with a high-resolution micro-CT at a resolution of 9 m.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 J Cell Mol Med. 2019 J