Pharmaceutical compositions for the treatment or prevention of inflammatory diseases
A composition and drug technology, applied in the direction of drug combination, allergic diseases, bone diseases, etc., can solve problems such as the unexplained anti-inflammatory mechanism of breviscapella
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Embodiment 1
[0089] Example 1: Preparation of Sopubia trifida Buch.-Ham.ex D.Don Extract
[0090] 1-1. Preparation of Grass brevis extract
[0091] Grass brevis is collected from the medicinal botanical garden of Shandong University of Traditional Chinese Medicine. Take dried and ground 10kg leaves of breves brevis, extract them with 95% ethanol for 3 times, each time for 5 hours, then filter and take the supernatant . The obtained supernatant was concentrated in vacuo to obtain 0.5 kg of ethanol extract of Brachyphyllum brevifolia.
[0092] 1-2. Preparation of Grass brevis fraction
[0093] The ethanol extract of Grass brevifolia prepared in Example 1-1 was suspended in 1 L of water, further mixed with 1 volume of 100% petroleum ether, and left to stand while stirring. This process was performed twice while removing the aqueous layer. As a result, 1.8 g of petroleum ether extract was obtained. After removing the petroleum ether fraction, an equal volume of chloroform was added and st...
Embodiment 2
[0094] Example 2: Inhibitory Activity Test of Grass brevis Extracts and Fractions on Cell Growth and NO Production
[0095] 2-1. Cell viability test
[0096] Mouse macrophage J774.1 was selected as the test cell, and the effect of different concentrations of Brachycarpa extract on cell viability was detected. More specifically, mouse macrophage J774.1 was treated with 1x10 6 Cells / ml were suspended in phenol red-free DMEM supplemented with 5% fetal bovine serum, and then the suspension was seeded in a 96-well cell culture plate at 80 μL / well. Four hours later, the adherent cells were incubated with each fraction for 24 hours and then incubated with 10 μL per well of 5 mg / ml MTT solution for an additional 4 hours. After removing the medium, add 100 μL of DMSO to each well and read the absorbance at 550 nm. Cell viability was expressed as a percentage of the absorbance at 550 nm relative to the negative control treated with 0.1% DMSO, calculated according to a mathematical for...
Embodiment 3
[0110] Example 3: Anti-inflammatory activity of Grass brevis extracts in J774.1 cells
[0111] 1-1. Inhibitory activity against NF-κB
[0112] The translocation of NF-κB into the nucleus of J774.1 cells under LPS-induced inflammatory conditions was detected by immunofluorescence. For this, J774.1 cells were grown at 1x10 6 Cells / ml were suspended in DMEM containing phenol red and 10% fetal bovine serum, seeded onto plastic chamber slides, and allowed to attach for 8 hours to the slide. Subsequently, the cells were incubated with 50 μg / mL of each Brachycarpa extract for 2 hours, and then further incubated with 1 μg / mL lipopolysaccharide (LPS, Sigma) for 1.5 hours. After removing the medium, the cells were fixed with ethanol for 30 min at 4°C, washed with phosphate-buffered saline, and blocked with 5% bovine serum albumin for 40 min at room temperature. Then, cells were reacted with primary antibody [anti-NF-κB antibody (1:100)] for 5 hours at room temperature, washed thoroug...
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