Application of ganoderma extracts and ganoderma triterpenes monomers for treating autosomal dominant inheritance polycystic kidney diseases
A Ganoderma lucidum extract, dominant genetic technology, applied in the field of biomedicine
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Embodiment 1
[0098] Embodiment 1, preparation of Ganoderma lucidum total triterpenes (Ganoderma lucidum extract)
[0099] 1. Preparation of Ganoderma lucidum total triterpenes (Ganoderma lucidum extract) and Ganoderma lucidum triterpene monomer
[0100]1.1 Preparation of Ganoderma lucidum total triterpenes (GT, Ganoderma lucidum extract)
[0101] After pulverizing 20kg of red lucidum fruiting body, heat and reflux extraction with 95% (volume percentage concentration) ethanol aqueous solution for 3 times. To dryness, recover the solvent under reduced pressure to obtain 1Kg of ethanol extract (paste).
[0102] Suspend the ethanol extract in 5L of water to obtain an ethanol extract suspension; add ethyl acetate that is 1 times the volume of the ethanol extract suspension to the ethanol extract suspension for extraction, and collect the ethyl acetate phase; the remaining liquid Continue to extract with ethyl acetate that is 1 times the volume of the remaining liquid, and extract with ethyl a...
Embodiment 2
[0135] Embodiment 2, Ganoderma lucidum total triterpenoids to the inhibitory action of MDCK vesicle formation and growth
[0136] 1. The total triterpenes of Ganoderma lucidum can inhibit the formation of MDCK vesicles without affecting the total number of cell colonies
[0137] Vesicle formation inhibition assay:
[0138] Canine kidney cells (Madin-Darby canine kidney cells, MDCK) were cultured in three-dimensional Matrigel (Purecol Collagen, Inamed Biomaterials Fremont Company, Cat. No. 5409) in vitro. The culture medium 1 used was to add three-dimensional Matrigel, HEPES, penicillin and streptomycin obtained the three-dimensional matrigel concentration of 2.9mg / ml, HEPES concentration of 10mM, penicillin concentration of 100U / ml, streptomycin concentration of 100μg / ml culture solution, pH 7.4. Culture solution 2 is a culture solution with FBS concentration of 10% and forskolin concentration of 10 μM by adding FBS and forskolin (FSK, forskolin, Sigma company, product number...
Embodiment 3
[0144] Example 3. Determination of the vesicle inhibitory effect of Ganoderma lucidum total triterpenoids in the kidney through an in vitro embryonic kidney model
[0145] In the evening of the first day, C57BL / 6 mice over 6 weeks old (Experimental Animal Center, Peking University Health Science Center) were mated with males and females in the same cage at a ratio of 1:1. The plug indicates that the female mouse has been pregnant for half a day. The mice without the vaginal plug are divided into cages first, and then closed in the cage at night, and then observed on the second day; the pregnant female mice are fed alone for 13 days, and the transwell plate is used for the embryonic kidney on the 13th day (Corning Company, Cat. No. 3401) for cultivation.
[0146] Take the 13.5-day-old mouse embryonic kidney and place it in the upper chamber of the transwell, and add 8-Br-cAMP (Sigma Company, Cat. No. B-5386) with a final concentration of 100 μM and 10% fetal bovine Serum, L-gl...
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