CN107296840B - Cape jasmine and fermented soybean soup effective part and application thereof in preparation of neuroprotective drugs - Google Patents
Cape jasmine and fermented soybean soup effective part and application thereof in preparation of neuroprotective drugs Download PDFInfo
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Abstract
The invention relates to the technical field of medicines, in particular to a method for separating and purifying an effective part with neuroprotective effect activity from gardenia and fermented soybean decoction, which comprises the following steps: soaking in water or ethanol water solution or refluxing to obtain crude product, eluting with small amount of water and ethanol water solution respectively, purifying with macroporous resin, collecting eluate, concentrating, and vacuum drying. The invention also relates to application of the gardenia and fermented soybean decoction effective part extract in preparing a neuroprotective medicament. The prepared gardenia fermented soybean decoction effective part can obviously inhibit the damage of PC12 cells induced by glutamic acid, prevent the release of LDH (layered double hydroxide) of the cells, improve the survival rate of the cells, reduce the oxygen free radical level in the cells and improve the activities of glutathione reductase and superoxide dismutase. The invention has simple preparation process, high yield, no environmental pollution and obvious drug effect, and is suitable for industrial production.
Description
Technical Field
The invention relates to the technical field of medicines, in particular to a part with neuroprotective activity prepared from a traditional Chinese medicine gardenia and fermented soybean soup and application of the part in preparing a neuroprotective medicine.
Background
With the continuous acceleration of the modern urban life rhythm, the pressure of people is higher and higher, the incidence rate of depression is increased year by year, and the depression becomes a mental disease seriously harming the physical and mental health of human beings at present. The world health organization predicts that depression will be the second most dangerous disease after heart disease in humans by 2020.
The Zhizi Chirong Tang is from Shang Han Lun (treatise on Cold-induced diseases) and comprises Zhi Zi and fermented soybean. In the formula, Gardenia is dried mature fruit of a madder plant Gardenia jasminoides ellis, and the main active component of the Gardenia is iridoid glycoside; in the formula, the semen Sojae Preparatum is fermented product of mature seed of Glycine max (L.) Merr of Leguminosae, and its main ingredient is soybean isoflavone compound. From the perspective of the theory of traditional Chinese medicine, domestic scholars perform a large number of syndrome differentiation treatment and formula syndrome analysis on the gardenia fermented soya beans and similar formulas thereof, and think that the gardenia is bitter and cold and has red color, the shape is similar to that of the gardenia, the red color is similar to that of the gardenia, the cold can clear heat, the bitter can clear and purge, the essence can clear heart and relieve restlessness, and the three-jiao fire can be purged; the blanched bugbane herb is pungent, sweet and slightly cold, floats and ascends slightly, transforms turbid into clear, nourishes yin, passes through heat, relieves exterior syndrome and eliminates restlessness. The two medicines are combined to promote the kidney water to reach upwards, dissolve and nourish yin, purge fire to nourish kidney yin, coordinate water and fire, coordinate yin and yang, and cause pathogenic heat to be self-contained, so that monarch, minister and self-contained medicine can be self-contained. Although the prescription is simple, the compatibility is precise and appropriate, and the meaning is infinite. Meanwhile, the clinical application of the gardenia fermented soybean decoction in the traditional Chinese medicine also shows that the gardenia fermented soybean decoction has good regulating and improving effects on a plurality of main symptoms of sub-health states such as anxiety, depression, insomnia, neurasthenia and the like.
Disclosure of Invention
The invention aims to provide a method for separating and purifying effective part groups with neuroprotective activity application potential from gardenia fermented soybean soup, which has the advantages of simple and convenient operation, low cost, high yield and purity and no environmental pollution.
In view of the fact that the gardenia fermented soybean decoction is a traditional Chinese medicine compound, but the extraction of antioxidants and corresponding active functional ingredients of the gardenia fermented soybean decoction are not very clear, most pharmacological experiments only stay on the basis of original medicinal materials or crude substances, and the gardenia fermented soybean decoction is not refined to an active part and is used for neuroprotection reports.
In order to test and evaluate the application potential of the active site of the gardenia fermented soybean decoction in the aspect of neuroprotection, the biological activity of each site of the medicinal material is tested, and the test method comprises the following steps: extracting and purifying active parts of the gardenia fermented soybean decoction, damaging PC12 cells by glutamic acid, and carrying out an antioxidant model and the like. The method comprises the following steps:
(1) preparing crude extracts of the gardenia and fermented soybean soup: mixing dried fructus Gardeniae and semen Sojae Preparatum coarse powder according to formula ratio, pulverizing, sieving with pharmacopeia No. 2 sieve, and preparing fructus Gardeniae and semen Sojae Preparatum decoction extract, mixing the powders (1:1, 2:1, 3:1 or 4:1), heating in an electric heating jacket at 100 deg.C, refluxing twice, each for two hours, mixing the extractive solutions, vacuum filtering, centrifuging at 3000r/min for 10min, collecting supernatant, concentrating the filtrate under reduced pressure to obtain extract, and vacuum drying to obtain fructus Gardeniae and semen Sojae Preparatum decoction crude extract;
(2) separation and purification: and D, adding a proper amount of water into the crude extract of the gardenia fermented soybean decoction prepared in the step A for suspension, and preparing 10g of raw medicinal materials into 100mL of water to prepare a solution equivalent to 1g/mL of the raw medicinal materials. Passing through macroporous adsorbent resin, eluting with a small amount of water until the eluent is not turbid, discarding, eluting with 10% ethanol, 20% ethanol, 30% ethanol, 40% ethanol, 50% ethanol, 60% ethanol, 70% ethanol, and 75% ethanol at a ratio of 1:1, collecting eluates, concentrating under reduced pressure, drying to obtain eluates with different concentrations, and vacuum drying to obtain powder.
(3) The MTT method screens the activity of each component medicament and screens out effective parts with obvious neuroprotective effect.
(4) The MTT and LDH method is used for measuring the cell activity, the intracellular oxygen free Radical (ROS) level, and the activity of glutathione reductase (GSH) and superoxide dismutase (SOD).
The invention inspects the optimal compatibility of the gardenia and fermented soybean soup, inspects the influence of different compatibility ratios of gardenia and fermented soybean on the decocting rate of chemical components, takes iridoid glycoside and soybean isoflavone as main index components, and adopts an ultraviolet spectrophotometry to determine the decocting amount of the two components in different compatibility ratios of a compound; the optimal compatibility ratio of the gardenia and fermented soybean soup is determined as follows: fermented soybean (weight ratio) is 2: 1.
Test results show that the gardenia and fermented soybean decoction macroporous resin 40% ethanol elution part has good neuroprotective activity with the administration concentration of 2 mg/mL.
Through repeated nerve protection activity screening of each part of the gardenia fermented soybean decoction extract, the extract eluted by the gardenia fermented soybean decoction macroporous resin with 40% ethanol is confirmed to have the effects of obviously inhibiting the damage of PC12 cells induced by glutamic acid, preventing LDH (layered double hydroxide) release of the cells, improving the survival rate of the cells, reducing the level of intracellular oxygen free Radicals (ROS), and improving the activities of glutathione reductase (GSH) and superoxide dismutase (SOD).
The invention provides a preparation method of an extract of an effective part of gardenia and fermented soybean soup, which comprises the following steps:
A) preparing crude extracts of the gardenia and fermented soybean soup: soaking dried gardenia fruits and fermented soybean coarse powder in water or extracting with ethanol under reflux for 1-3 times according to the weight ratio of 1: 5-5: 1, filtering, concentrating the filtrate under reduced pressure to obtain a paste, and then drying in vacuum to obtain a crude extract of gardenia fermented soybean soup;
B) separation and purification: and D, adding water into the crude extract of the gardenia fermented soybean soup prepared in the step A for suspension, passing through macroporous adsorption resin, eluting with a small amount of water until the eluent is not turbid, discarding the eluent, eluting with 10-75% ethanol, collecting the eluent, and concentrating and drying under reduced pressure to obtain the extract of the effective parts of the gardenia fermented soybean soup.
Preferably, in the step A, the weight ratio of the gardenia to the fermented soybean is 1:1, 1:2 or 2: 1.
Preferably, in the step a, when the crude extract of the gardenia and fermented soybean decoction is prepared, the concentration of ethanol used for reflux extraction is 1-100%. More preferably, the ethanol concentration used in the reflux extraction is 50%.
Preferably, in the step B, the volume of water in the elution of a small amount of water is not more than 1 time of the resin amount.
Preferably, in the step B, the elution is carried out by using 30-60% ethanol. More preferably, the elution is carried out with 40% strength ethanol.
Preferably, in the step B, the macroporous adsorption resin is 1300 or D101 or AB-8 type macroporous adsorption resin.
In a second aspect of the invention, the gardenia and fermented soybean decoction effective part extract prepared by the method is provided.
The third aspect of the invention provides an application of the extract of the effective parts of the cape jasmine and fermented soybean soup in preparing a neuroprotective medicament.
Preferably, the concentration of the cape jasmine and fermented soybean decoction effective part extract in the neuroprotective medicament is 1-3 mg/mL. More preferably 2 mg/mL.
The invention has the beneficial effects that:
1. a small amount of water is used for eluting firstly, so that part of water-soluble impurities can be washed away, and the loss of active ingredients with strong polarity can be reduced, thereby improving the yield;
2. the invention simultaneously compares the drug effects of the active sites of the gardenia fermented soybean decoction macroporous resin ethanol extract in different proportions, preferably selects the gardenia fruit and the fermented soybean coarse powder in the gardenia fermented soybean decoction to be mixed and extracted according to the weight ratio of 2:1, and the active sites have obvious neuroprotective effect after being eluted by macroporous resin 40% ethanol;
3. the invention adopts macroporous adsorption resin to separate and purify the effective part group with the neuroprotective effect in the gardenia fermented soybean soup, has simple preparation process, high yield, no environmental pollution and very obvious drug effect, and is suitable for industrial production.
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FIG. 1 is a process flow diagram of the present invention.
Detailed Description
The following examples are provided to illustrate specific embodiments of the present invention.
Example 1
(1) Preparing crude extracts of the gardenia and fermented soybean soup: mixing 40g of dried gardenia fruits and 20g of fermented soybean coarse powder, crushing the mixture, sieving the mixture by a No. 2 pharmacopoeia sieve to prepare a gardenia fermented soybean soup extract, mixing the gardenia fermented soybean soup extract and the fermented soybean soup extract, placing the mixture into a round-bottom flask, carrying out reflux extraction for 2 times by using 50 percent ethanol in an amount which is 8 times that of the mixture, carrying out filtration for 60min each time, combining filtrates, carrying out reduced pressure concentration at 60 ℃ to obtain a thin extract, and carrying out vacuum drying to obtain the gardenia fermented.
(2) And (3) purification: taking 10g of the crude extract of the gardenia and fermented soybean decoction, and adding water to suspend, wherein the total volume is 100 ml. Adding into a chromatographic column filled with 100ml of pretreated D101 macroporous adsorption resin, and adsorbing for 2 hours after the sample loading is finished. Elute with 100ml water (discard). Eluting with 100ml 10% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 1.26% yield, eluting with 100ml 20% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 2.91% yield, eluting with 100ml 30% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 2.62% yield, eluting with 100ml 40% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 2.46% yield, eluting with 100ml 50% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 1.92% yield, eluting with 100ml 60% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 0.86% yield, eluting with 100ml 70% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 0.C yield, the yield is 0.48%, then 100ml of 75% ethanol is used for elution, the ethanol elution fraction is decompressed and concentrated, and then vacuum drying is carried out (60 ℃), and the yield is 0.09%.
(3) Determination of intracellular MTT and LDH activity
About 5X 10 cells per well were collected from PC12 cells in logarithmic growth phase4And inoculating the cells in a 96-well culture plate, and dividing the cells into a normal group, a model group and a dosing group after the cells are attached overnight, wherein each group is provided with 6 multiple wells. Normal cell Control (Control) (complete medium); ② a model group (complete medium +5mmol/L Glu); ③ administration group (complete culture medium +5mmol/L Glu + 40% ethanol extract active site of gardenia fermented soybean decoction, final concentration is 2 mg/mL); placing at 37 ℃ and 5% CO2After 24h of incubation in an incubator, 20 μm was added to each wellMTT solution with the L final concentration of 5mg/mL is incubated in an incubator for 4h, supernatant in the holes is carefully discarded, 150 mu L of dimethyl sulfoxide (DMSO) is added into each hole, a horizontal shaking table is oscillated for 10min, after the purple blue particles in the holes are completely dissolved, the optical density (OD value) of each hole is read by a microplate reader at the wavelength of 570nm, and the cell survival rate is calculated according to the formula: survival rate = experimental group OD value/normal group OD value × 100%; the results are shown in table 1, and the data show that the gardenia and fermented soybean decoction effective part extract can protect the injury effect of PC12 cells.
Cell supernatants were collected at 100. mu.L and assayed for LDH (Dojindo, Inc. purchased in Japan) activity according to the kit instructions. PC12 cells from the blank wells were allowed to release all LDH using cell lysates as a reference total concentration of LDH. The formula is cytotoxicity ═ a-C)/(B-C) × 100, a is the test substance, B is total LDH release, and C is the blank. The results are shown in table 2, and the data show that the extract of the effective parts of the gardenia and fermented soybean soup has the effect of inhibiting Glu induced LDH release of PC12 cells.
Table 1 MTT assay of influence of gardenia fermented soya bean soup on survival rate of Glu-induced impaired PC12 cells
###p<The normal group of 0.001vs,***p<0.001vs model set.
TABLE 2 LDH determination of influence of cape jasmine fermented soya bean decoction on survival rate of Glu-induced impaired PC12 cells
###p<0.001vs normal group<0.001vs model set.
(4) Determination of intracellular ROS levels, GSH and SOD Activity
Taking PC12 cells in logarithmic growth phase, and adding 5 × 104The cells were seeded in 6-well plates at 37 ℃ in 5% CO2After overnight incubation under conditions, the assay was performed according to reactive oxygen species ROSThe operation of the test kit is guided and divided into a normal group, a model group and an administration group, and each group is provided with three multiple holes. After further culturing for 24 hours, collecting cells, gently washing the cells twice by PBS, adding DCFH-DA diluted without serum to enable the final concentration to be 10 mu M, uniformly blowing and beating the cells, after incubating the cells in an incubator at 37 ℃ for 20min, washing the cells three times by a serum-free culture medium, fully removing probes not loaded in the cells, adding 300 mu L PBS to resuspend the cells, and detecting the fluorescence intensity by a flow cytometer. The results are shown in table 3, and the data show that the extract of the effective part of the gardenia and fermented soybean soup can obviously inhibit the ROS (reactive oxygen species) production of PC12 cells induced by glutamic acid.
Taking PC12 cells in logarithmic growth phase, and adding 5 × 104The cells were seeded in Petri dishes at a density of 5% CO at 37 ℃%2After overnight culture under the condition, the culture medium is divided into a normal group, a model group and an administration group, and each group is provided with three multiple holes. And removing the supernatant after 24h, washing twice with PBS, reserving 1mL of PBS, gently scraping the cells with a cell scraper, collecting the cells in a 1.5mL centrifuge tube at 1500rpm for 5min, sucking the supernatant, adding cell lysate to lyse the cells, lysing the cells for 15min at 13000rpm, centrifuging the cells for 15min, taking the supernatant, measuring the protein concentration with a BCA kit, performing reaction of each step according to the operation guidance of the detection kit, and respectively measuring the OD value of each group of cells with an enzyme labeling instrument. The results are shown in table 4, and data show that the extract of the effective part of the gardenia and fermented soybean soup can obviously improve the activity of glutathione reductase (GSH) and superoxide dismutase (SOD).
TABLE 3 flow cytometry for determining the influence of the gardenia fermented soya bean soup on Glu-induced PC12 cell ROS
###p<0.001vs normal group<0.001vs model set.
TABLE 4 enzyme-labeling instrument for determining activity of cape jasmine and fermented soybean soup on Glu-induced PC12 cell GSH and SOD
##p<0.01 vs. Normal group,. p<0.01vs model set.
Example 2
(1) Preparing crude extracts of the gardenia and fermented soybean soup: mixing 40g of dried gardenia fruits and 20g of fermented soybean coarse powder, crushing the mixture, sieving the mixture by a No. 2 pharmacopoeia sieve to prepare a gardenia fermented soybean soup extract, mixing the gardenia fermented soybean soup extract and the fermented soybean soup extract, placing the mixture into a round-bottom flask, carrying out reflux extraction for 2 times by using 50 percent ethanol in an amount which is 8 times that of the mixture, carrying out filtration for 60min each time, combining filtrates, carrying out reduced pressure concentration at 60 ℃ to obtain a thin extract, and carrying out vacuum drying to obtain the gardenia fermented.
(2) And (3) purification: and (3) purification: taking 10g of the crude extract of the gardenia and fermented soybean decoction, and adding water to suspend, wherein the total volume is 100 ml. Adding into a chromatographic column filled with 100ml of pretreated D101 macroporous adsorption resin, and adsorbing for 2 hours after the sample loading is finished. Elute with 100ml water (discard). Eluting with 100ml 10% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 1.26% yield, eluting with 100ml 20% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 2.91% yield, eluting with 100ml 30% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 2.62% yield, eluting with 100ml 40% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 2.46% yield, eluting with 100ml 50% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 1.92% yield, eluting with 100ml 60% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C) to obtain 0.86% yield, eluting with 100ml 70% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C), and eluting with 100ml 75% ethanol, concentrating the ethanol eluate under reduced pressure, and vacuum drying (60 deg.C) to obtain 0.48% yield.
(3) Determination of intracellular MTT and LDH activity
About 5X 10 cells per well were collected from PC12 cells in logarithmic growth phase4And inoculating the cells in a 96-well culture plate, and dividing the cells into a normal group, a model group and a dosing group after the cells are attached overnight, wherein each group is provided with 6 multiple wells. Normal cell Control (Control) (complete medium); ② model group (complete culture medium +5mmol/lGlu); ③ administration group (complete culture medium +5mmol/L Glu + 30% ethanol extract active site of gardenia fermented soybean soup macroporous resin, final concentration is 3 mg/mL); the detection method was the same as in example 1.
TABLE 5 MTT assay of the Effect of cape jasmine and fermented soybean decoction on the survival rate of Glu-induced impaired PC12 cells
###p<The normal group of 0.001vs,***p<0.001vs model set.
TABLE 6 LDH determination of influence of cape jasmine fermented soya bean decoction on survival rate of Glu-induced impaired PC12 cells
###p<0.001vs normal group<0.001vs model set.
(4) Measurement of intracellular ROS levels, GSH and SOD Activity.
The detection method was the same as in example 1.
TABLE 7 flow cytometry for determining influence of gardenia black bean soup on Glu-induced PC12 cell ROS
###p<0.001vs normal group<0.001vs model set.
TABLE 8 enzyme-labeling instrument for determining activity of cape jasmine and fermented soybean soup on Glu-induced PC12 cell GSH and SOD
##p<0.01 vs. Normal group,. p<0.01，***p<0.001vs model set.
Example 3
(1) Preparing crude extracts of the gardenia and fermented soybean soup: mixing 30g of dried gardenia fruits and 30g of fermented soybean coarse powder, crushing the mixture, sieving the mixture by a pharmacopoeia No. 2 sieve to prepare a gardenia fermented soybean decoction extract, mixing the gardenia fermented soybean decoction extract and the fermented soybean decoction extract, placing the mixture into a round-bottom flask, carrying out reflux extraction for 2 times by using 50 percent ethanol in an amount which is 8 times that of the total amount of the mixture, carrying out 60min each time, filtering, combining filtrates, carrying out reduced pressure concentration at 60 ℃ to obtain a thin extract, and carrying out vacuum drying to obtain the.
(2) And (3) purification: taking 10g of the crude extract of the gardenia and fermented soybean decoction, and adding water to suspend, wherein the total volume is 100 ml. Adding into a chromatographic column filled with 100ml of pretreated D101 macroporous adsorption resin, and adsorbing for 2 hours after the sample loading is finished. Elute with 100ml water (discard). Eluting with 100ml 10% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C), the yield is 1.26%, eluting with 100ml 20% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C), the yield is 1.31%, eluting with 100ml 30% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C), the yield is 1.65%, eluting with 100ml 40% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C), the yield is 1.86%, eluting with 100ml 50% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C), the yield is 1.24%, eluting with 100ml 60% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C), the yield is 0.74%, eluting with 100ml 70% ethanol, concentrating the ethanol eluate under reduced pressure, vacuum drying (60 deg.C), the yield is 0.43%, then 100ml of 75% ethanol is used for elution, and the ethanol elution fraction is decompressed, concentrated and dried in vacuum (60 ℃), and the yield is 0.24%.
(3) Determination of intracellular MTT and LDH activity
About 5X 10 cells per well were collected from PC12 cells in logarithmic growth phase4And inoculating the cells in a 96-well culture plate, and dividing the cells into a normal group, a model group and a dosing group after the cells are attached overnight, wherein each group is provided with 6 multiple wells. Normal cell Control (Control) (complete medium); ② a model group (complete medium +5mmol/l Glu); ③ administration setExtracting active parts from the total culture medium, 5mmol/L Glu, 60% ethanol of gardenia and fermented soybean decoction macroporous resin, wherein the final concentration is 1 mg/mL); the detection method was the same as in example 1.
TABLE 9 MTT assay of the Effect of cape jasmine and fermented soybean decoction on the survival rate of Glu-induced impaired PC12 cells
###p<0.001vs normal group.
TABLE 10 LDH determination of Effect of cape jasmine and fermented soya bean decoction on survival Rate of Glu-induced impaired PC12 cells
###p<0.001vs normal group<0.001vs model set.
(4) Measurement of intracellular ROS levels, GSH and SOD Activity.
The detection method was the same as in example 1.
TABLE 11 flow cytometry for determining the influence of cape jasmine and fermented soybean soup on Glu-induced PC12 cell ROS
###p<0.001vs normal group<0.001vs model set.
TABLE 12 enzyme-labeling instrument for determining activity of cape jasmine and fermented soybean soup on Glu-induced PC12 cell GSH and SOD
##p<0.01 vs. Normal group,. p<0.01vs model set.
While the preferred embodiments of the present invention have been described in detail, it will be understood by those skilled in the art that the invention is not limited thereto, and that various changes and modifications may be made without departing from the spirit of the invention, and the scope of the appended claims is to be accorded the full range of equivalents.
Claims (2)
1. Application of an extract of the effective part of cape jasmine and fermented soybean decoction in preparing neuroprotective drugs; the preparation method of the extract of the effective part of the gardenia and fermented soybean decoction comprises the following steps:
A) preparing crude extracts of the gardenia and fermented soybean soup: extracting the dried gardenia fruits and the fermented soybean coarse powder for 1-3 times by ethanol in a weight ratio of 2:1, wherein the concentration of the ethanol used for reflux extraction is 50%; filtering, concentrating the filtrate under reduced pressure to obtain extract, and vacuum drying to obtain crude extract of fructus Gardeniae and fermented soybean decoction;
B) separation and purification: adding water into the crude extract of the gardenia fermented soybean decoction prepared in the step A for suspension, passing through macroporous adsorption resin which is 1300 or D101 or AB-8 type macroporous adsorption resin, eluting with a small amount of water until the eluent is not turbid, and discarding the water with the volume not more than 1 time of the resin amount during the elution of the small amount of water; sequentially eluting with 10%, 20%, 30%, 40%, 50%, 60%, 70% and 75% ethanol, collecting eluate, concentrating under reduced pressure, and drying to obtain the product of the effective component extract of cape jasmine and fermented soybean soup.
2. The application of the gardenia and fermented soybean decoction effective part extract in the preparation of the neuroprotective medicament according to claim 1, wherein the medicament concentration of the gardenia and fermented soybean decoction effective part extract in the neuroprotective medicament is 1-3 mg/mL.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104147183A (en) * | 2014-07-18 | 2014-11-19 | 中国人民解放军第二军医大学 | Method for separating and purifying total iridoid glycosides and total soy isoflavones from capejasmine and fermented soybean decoction |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104147183A (en) * | 2014-07-18 | 2014-11-19 | 中国人民解放军第二军医大学 | Method for separating and purifying total iridoid glycosides and total soy isoflavones from capejasmine and fermented soybean decoction |
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| dFGEN对谷氨酸诱导的PC12细胞损伤的保护作用研究;陶美皊;《医药卫生科技辑》;20130115(第1期);E079-196，尤其是第21页表6、第23页表7和第28页表9 * |
| 大豆异黄酮活性组分对H2O2诱导的PC12细胞损伤的影响;陈伟强等;《中国临床药理学与治疗学》;20050430;第10卷(第4期);第443-446页，尤其是第443页摘要 * |
| 大豆苷元对谷氨酸体外诱导海马神经元损伤大鼠的保护作用;秦传勇;《医学导报》;20100131;第29卷(第1期);第25-27页，尤其是第26页右栏第1段和第27页左栏第3段 * |
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| 栀子豉汤活性成分分离制备的多维色谱技术研究;方高;《医药卫生科技辑》;20140715(第7期);E057-44，尤其是第58页第4段、第59页第2段和第60页第1段 * |
| 胰高血糖素样肽1受体激动的高通量筛选及作用机制研究;殷菲等;《中国药学杂志》;20070131;第42卷(第1期);第24-27页，尤其是第27页左栏第1、3段 * |
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