Chinese traditional extraction and the function containing dammarane type four-ring triterpene sapogenin
A tetracyclic triterpenoid and dammarane-type technology, applied in the field of dammarane-type tetracyclic triterpene saponin extract, can solve problems such as protein function destruction, calcium ion overload, necrosis, etc., and achieve the goal of reducing cardiotoxicity Effect
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Embodiment 1
[0016] Take 500g of Panax notoginseng medicinal material granules (3mm-8mm), dry at 50°C for 6 hours, add 10 times the amount of 70% ethanol and heat and reflux in a slightly boiling state to extract twice, each time for 2 hours, filter the extract, combine, and concentrate under reduced pressure to 0.5 g / mL (crude drug amount), above D 101 Macroporous resin. Rinse with 4 times the column volume of deionized water, discard, then rinse with 4BV of 0.05% ammonia water, discard, then rinse with deionized water until the effluent pH=7, continue to rinse with 4BV of 30% ethanol solution, discard, Finally, it was eluted with 5BV 70% ethanol solution, and the eluate was collected, concentrated under reduced pressure to no alcohol, and freeze-dried to obtain the total saponins of Panax notoginseng. Determined by HPLC external standard method, notoginseng saponin R in the refined product 1 8%, Ginsenoside Rg 1 44%, Ginsenoside Rb 1 28%, ginsenoside Rd10%, ginsenoside Re5%, the tota...
Embodiment 2
[0018] The notoginseng medicinal material is crushed to the size of soybeans to coarse powder, extracted 4 times with 40% water-containing alcohol under reflux, recovered the solvent, extracted 3 times with an equal volume of n-butanol, combined the extracts, concentrated to dryness, crushed, and washed with petroleum ether for 2 times, desolvation and drying. Adopt high performance liquid phase method (HPLC) to detect it, analysis condition is Agilent Extend-C18, mobile phase H 2 O-CH3CN, the detection wavelength is 203nm, and its content is determined by the external standard method. The contents of the five components are respectively notoginseng saponin R 1 3%, Ginsenoside Rg 1 25%, Ginsenoside Rb 1 14%, ginsenoside Rd4%, ginsenoside Re1.5%. The total content is 47.5%.
Embodiment 3
[0020] Cardiomyocytes from neonatal rats (Wistar rats) were isolated and seeded into 96-well plates. Cell density 5×10 5 pcs / hole. After culturing in DMEM+20% FBS medium for 48 hours, the cells adhered to the wall normally, and some cells beat regularly. Set up control group, model group (containing 0.1, 1 μ M doxorubicin hydrochloride), and different concentrations (12.5-100 μ g / ml) of notoginseng saponin group (added 1 hour before adding doxorubicin). After culturing for 24 hours, it was measured by MTT method, and the viability of cardiomyocytes was reflected by the 490nm colorimetric value. See results figure 1 (**P<0.01; *P<0.05).
[0021] Experiments have shown that 0.1 and 1 μM doxorubicin hydrochloride can cause significant myocardial cell damage, and the MTT value is significantly reduced; while the pre-administration of notoginseng saponin at a concentration of 12.5-100 μg / ml shows a certain protective effect. It shows that notoginseng saponins can prevent and a...
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