Composition for prevention and treatment of bone disease containing mixed Chinese herbal medicine extract
A composition and extract technology, which can be applied in the direction of drug combination, food ingredients containing natural extracts, medical preparations containing active ingredients, etc., can solve the problem of inability to present the therapeutic effect of osteoporosis
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Embodiment 1
[0056] Embodiment 1. Preparation of mixed crude medicinal material extract
[0057] 1-1. Raw medicinal material screening and pretreatment process
[0058] For 150g of Chinese Angelica, 150g of Radix Paeoniae Alba, 150g of Atractylodes Rhizome, 150g of Poria, 100g of Gardenia, 100g of Licorice, 100g of Bellflower, 50g of Dried Ginger and 50g of Peppermint, after removing foreign matter, finely cut them into about 5cm. Extracts easily and rinses cleanly with water. The purpose of finely chopping raw herbs is to facilitate their handling and extraction.
[0059] 1-2. Preparation of hot water extract of mixed raw medicinal materials
[0060] The material obtained in the above-mentioned process is put into the extraction device, and extracted with an extraction solvent. As an extraction solvent, pure water was added 5 times as much as the crude drug, and extracted at a temperature of 95° C. for 6 hours with a cooling extraction device. The extract was soaked naturally for 1 to...
Embodiment 2
[0072] Example 2. Activity evaluation of osteoblasts
[0073] 2-1. Cell culture and differentiation induction
[0074] MC3T3-E1 osteoblast cells (MC3T3-E1 osteoblastic cells) derived from raw mice were purchased from American Type Culture Collection (ATCC, American Type Culture Collection). MC3T3-E1 cells were prepared by adding 10% fetal bovine serum (FBS, fetal bovine serum) and 100 U / mL penicillin ( penicillin), 100U / mL of streptomycin (streptomycin) in cell culture medium at 37°C in humidified CO 2 Incubator (5% CO 2 / 95% air (air)). If the cells fill up about 80% of the culture dish, then utilize phosphate buffered saline (PBS, pH7.4) to wash the monolayer of cells, and process 0.25% trypsin (trypsin)-2.65mM EDTA ( EDTA) for subculture, and the medium was changed every 2 days.
[0075] When MC3T3-E1 cells filled up about 90% of the culture dish, in order to induce cell differentiation, β-glycerophosphate (β-glycerophosphate) (Sigma-Aldrich) (Sigma- Aldrich Co.)) and...
Embodiment 3
[0086] Example 3. Activity evaluation of osteoclasts
[0087] 3-1. Cell culture and differentiation induction
[0088] Raw264.7 cells were purchased from the American Type Culture Collection (ATCC, American Type Culture Collection) as macrophages derived from raw mice. Raw264.7 cells were prepared by adding 10% fetal bovine serum, 100units / mL of penicillin, and 100ug / mL of streptomycin in Dulbecco's modified Eagle's medium (DMEM, dulbecco's modified eagle medium, Welgene Company). primed cell culture medium at 37 °C in a humidified CO 2 Incubator (5% CO 2 / 95% air (air)). If the cells fill up about 80% of the culture dish, wash the monolayer of cells with phosphate buffered saline (PBS, pH 7.4), add cell culture medium to remove the cells for subculture, and replace them every 2 days Medium.
[0089] To induce the differentiation of Raw264.7 cells, 50 ng / mL of receptor activator ligand (RANKL) (Sigma-Aldrich Co.) and 10 μM of PD98059 were added to α-minimum essential medi...
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