CN115025141B - White dorsum Shemu total saponins and preparation method and application thereof - Google Patents
White dorsum Shemu total saponins and preparation method and application thereof Download PDFInfo
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
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- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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Abstract
The invention discloses a white dorsum Shemu total saponin and a preparation method and application thereof. The total saponins of the white back Shemu are total saponins of the young leaf buds of the white back Shemu. The preparation method of the total saponins of the white back Shemu comprises the steps of crushing the dried white back Shemu tender leaf buds, extracting with alcohol, concentrating to obtain an alcohol extract, suspending with water, and extracting sequentially with petroleum ether, ethyl acetate and n-butanol; concentrating the n-butanol extraction part, performing silica gel column chromatography, eluting with chloroform-methanol=95:5, and sequentially eluting with chloroform: methanol: water = 7:3:0.5 elution, methanol: water = 9:1 elution, chloroform was collected: methanol: concentrating the water=7:3:0.5 part under reduced pressure to obtain the total saponins of the white back Shemu. The dorsum albuminosus Shemu total saponins can obviously improve colon inflammation of mice induced by dextran sodium sulfate, have obvious activity of resisting ulcerative colitis, and have good application prospects in preparing medicines for treating inflammatory bowel diseases.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a white dorsum Shemu total saponin and a preparation method and application thereof.
Background
Shemu of white backAralia chinensis L. var. nudaNakai) is a plant of aralia genus of Araliaceae family, and has wide distribution in south and north of China and more distribution in Yunnan. The Chinese toon head, the root of Chinese thorn and the old bag are also called as the Chinese toon head, the root of Chinese thorn head and the old bag in the folk of Yunnan, and the tender stem and leaf is taken as the wild vegetable for eating, and has the effects of clearing heat and detoxicating. It is also commonly used for treating jaundice due to damp-heat, dysentery, enteritis, mastitis, rheumatalgia, etc. by taking root or root bark, stem bark, leaf, tender leaf bud, etc.
Currently, there are few reports about the extraction of the main active ingredient of white back Shemu. The invention aims to provide total saponins extracted from the buds of white back Shemu and application thereof.
Disclosure of Invention
The first object of the invention is to provide a total saponin of white dorsum Shemu, the second object of the invention is to provide a preparation method of the total saponin of white dorsum Shemu, and the third object of the invention is to provide an application of the total saponin of white dorsum Shemu.
The first object of the present invention is achieved in that the total saponins of the white back Shemu are total saponins extracts of young leaf buds of the white back Shemu.
The second purpose of the invention is realized by the preparation method of the total saponins of the white back Shemu, which comprises the following steps:
1) Pulverizing dried leaf bud of white back Shemu, extracting with alcohol, and concentrating to obtain alcohol extract;
2) Suspending the alcohol extract with water, sequentially extracting with petroleum ether, ethyl acetate and n-butanol for 2-5 times, and mixing the extracts;
3) Concentrating the n-butanol extraction part, performing silica gel column chromatography, eluting with chloroform-methanol=95:5, and sequentially eluting with chloroform: methanol: water = 7:3:0.5 elution, methanol: water = 9:1 elution, chloroform was collected: methanol: concentrating the water=7:3:0.5 part under reduced pressure to obtain the total saponins of the white back Shemu.
The third aspect of the present invention is realized in that the application of the total saponins of the white back Shemu is the application in preparing medicines for treating inflammatory bowel diseases.
The beneficial effects of the invention are as follows:
the white dorsum Shemu total saponins can obviously improve colon inflammation of mice induced by dextran sodium sulfate, and obviously reduce the activity index of the mice diseases and obviously improve the colon length shortening of the mice caused by modeling; significantly reducing MPO viability in colon tissue; the method has the advantages of remarkably reducing the content of inflammatory factors IL-1 beta and TNF-alpha in colon tissues, relieving inflammatory response of colon parts, remarkably resisting the activity of ulcerative colitis, and having good application prospect in preparing medicines for treating inflammatory bowel diseases.
FIG. 1 is a graph (200X) showing the effect of total saponins of white back Shemu of the present invention on DSS-induced colon histopathological changes in mice with UC model.
Detailed Description
The invention is further described below without limiting it in any way, and any modifications based on the invention fall within the scope of protection of the invention.
The invention provides a total saponin of white back Shemu, wherein the total saponin of white back Shemu is a total saponin extract of young leaf buds of white back Shemu.
The invention further provides a preparation method of the white dorsum Shemu total saponins, which is realized by the following steps:
1) Pulverizing dried leaf bud of white back Shemu, extracting with alcohol, and concentrating to obtain alcohol extract;
2) Suspending the alcohol extract with water, sequentially extracting with petroleum ether, ethyl acetate and n-butanol for 2-5 times, and mixing the extracts;
3) Concentrating the n-butanol extraction part, performing silica gel column chromatography, eluting with chloroform-methanol=95:5, and sequentially eluting with chloroform: methanol: water = 7:3:0.5 elution, methanol: water = 9:1 elution, chloroform was collected: methanol: concentrating the water=7:3:0.5 part under reduced pressure to obtain the total saponins of the white back Shemu.
In the step 1, the granularity of crushing the buds of the white back Shemu is 20-60 meshes;
the alcohol is methanol or ethanol, the concentration range of the ethanol is 60-85%, the concentration range of the methanol is 60-85%, and the dosage ratio of the medicinal material amount to the methanol or ethanol is as follows: 1kg/2L and 1kg/4L by reflux extraction, decoction or leaching.
The reflux extraction temperature is 85-100 ℃, the extraction time is 30-60 min each time, and the extraction times are 2-4 times.
In the step 2, when the alcohol extract is suspended by adding water, the dosage ratio of the alcohol extract to the water is as follows: 1g/4 mL-1 g/10mL.
In the step 2, the medium used for column chromatography of the n-butanol extraction part is 200-300 mesh silica gel.
The invention further provides application of the white dorsum Shemu total saponins in preparation of medicines for treating inflammatory bowel diseases.
The inflammatory bowel disease is ulcerative colitis.
The invention further provides a pharmaceutical composition comprising a therapeutically effective amount of the total saponins of dorsum total Shemu of the present invention as an active agent together with a pharmaceutically effective carrier.
The medicine is pill, capsule, tablet, powder, granule, injection, compound preparation or other oral dosage forms.
The total saponins of the white back Shemu are dosed 1-2 times per day, 150-300 mg each time.
The total saponins of the white back Shemu are dosed 2 times daily, 200 mg each time.
EXAMPLE 1 preparation of white dorsum Shemu Total saponins
Weighing the white back Shemu, drying the tender leaf bud 5kg, crushing into coarse powder of 20 meshes, heating and reflux-extracting with 70% ethanol at 90 ℃ for 30 min (extracting with 10L of solvent for 4 times each time), and concentrating under reduced pressure on a rotary evaporator to obtain 1.2 kg ethanol total extract. Suspending the ethanol total extract with 5L water, and sequentially extracting with equal volume of petroleum ether (3 times of extraction), ethyl acetate (3 times of extraction), and n-butanol (3 times of extraction) to obtain extractive solution. Concentrating the extractive solution under reduced pressure to obtain n-butanol extract which is total saponin enriched. Loading the n-butanol extraction part onto 300 mesh silica gel column, eluting with chloroform, chloroform-methanol (95:5), chloroform-methanol-water (7:3:0.5), methanol-water (9:1), and concentrating the chloroform-methanol-water (7:3:0.5) eluate under reduced pressure to obtain 456 g white back Shemu total saponins.
EXAMPLE 2 preparation of white dorsum Shemu Total saponins
Weighing the white back Shemu, drying the tender leaf bud 5kg, crushing into 40-mesh coarse powder, heating and reflux-extracting with 80% ethanol at 85deg.C for 40 min (each time with 12L solvent for 4 times), and concentrating under reduced pressure on a rotary evaporator to obtain 1.3kg ethanol total extract. Suspending the ethanol total extract with 6.5L water, and sequentially extracting with petroleum ether (2 times of extraction), ethyl acetate (2 times of extraction), and n-butanol (2 times of extraction) to obtain extractive solution. Concentrating the extractive solution under reduced pressure to obtain n-butanol extract which is total saponin enriched. Loading the n-butanol extraction part onto 200 mesh silica gel column, eluting with chloroform, chloroform-methanol (95:5), chloroform-methanol-water (7:3:0.5), methanol-water (9:1), and concentrating the chloroform-methanol-water (7:3:0.5) eluate under reduced pressure to obtain 403g of total saponins of radix Cynanchi auriculati Shemu.
EXAMPLE 3 preparation of white dorsum Shemu Total saponins
Weighing the white back Shemu, drying the tender leaf bud 5kg, crushing into 60-mesh coarse powder, heating and reflux-extracting with 65% ethanol at 100deg.C for 60min (each time with 14L solvent for 2 times), and concentrating under reduced pressure on a rotary evaporator to obtain 1.5kg ethanol total extract. Suspending the ethanol total extract with 7.5L water, and sequentially extracting with petroleum ether (4 times of extraction), ethyl acetate (4 times of extraction), and n-butanol (4 times of extraction) to obtain extractive solution. Concentrating the extractive solution under reduced pressure to obtain n-butanol extract which is total saponin enriched. Loading the n-butanol extraction part onto 300 mesh silica gel column, eluting with chloroform, chloroform-methanol (95:5), chloroform-methanol-water (7:3:0.5), methanol-water (9:1), and concentrating the chloroform-methanol-water (7:3:0.5) eluate under reduced pressure to obtain 424 g white back Shemu total saponins.
EXAMPLE 4 preparation of white dorsum Shemu Total saponins
Weighing white back Shemu, drying tender leaf bud 5kg, pulverizing into 40 mesh coarse powder, reflux-extracting with 80% methanol at 85deg.C for 60min (20L solvent for 2 times each time), concentrating under reduced pressure on rotary evaporator to obtain 1.1kg methanol extract, suspending with 10L water, and sequentially extracting with equal volume of petroleum ether (3 times of extraction), ethyl acetate (2 times of extraction), and n-butanol (4 times of extraction) to obtain extractive solution. Concentrating the extractive solution under reduced pressure to obtain n-butanol extract which is total saponin enriched. Loading the n-butanol extraction part onto 300 mesh silica gel column, eluting with chloroform, chloroform-methanol (95:5), chloroform-methanol-water (7:3:0.5), methanol-water (9:1), and concentrating the chloroform-methanol-water (7:3:0.5) eluate under reduced pressure to obtain 394g of total saponins of radix Cynanchi auriculati Shemu.
EXAMPLE 5 preparation of white dorsum Shemu Total saponins
Weighing white back Shemu, drying tender leaf bud 5kg, pulverizing into 40 mesh coarse powder, reflux-extracting with 65% methanol at 90deg.C for 45 min (each time with 16L solvent for 3 times), concentrating under reduced pressure on rotary evaporator to obtain 1.0kg methanol extract, suspending with 6L water, and sequentially extracting with equal volume of petroleum ether (3 times of extraction), ethyl acetate (2 times of extraction), and n-butanol (4 times of extraction) to obtain extractive solution. Concentrating the extractive solution under reduced pressure to obtain n-butanol extract which is total saponin enriched. Loading the n-butanol extraction part onto 300 mesh silica gel column, eluting with chloroform, chloroform-methanol (95:5), chloroform-methanol-water (7:3:0.5), methanol-water (9:1), and concentrating the chloroform-methanol-water (7:3:0.5) eluate under reduced pressure to obtain 388g of total saponins of radix Cynanchi auriculati Shemu.
EXAMPLE 6 preparation of white dorsum Shemu Total saponins
Weighing white back Shemu, drying tender leaf bud 5kg, pulverizing into 40 mesh coarse powder, extracting with 75% methanol under reflux at 98deg.C for 30 min (each time with 18L solvent for 4 times), concentrating under reduced pressure on rotary evaporator to obtain 0.9kg methanol extract, suspending with 7.2L water, and sequentially extracting with equal volume petroleum ether (3 times of extraction), ethyl acetate (2 times of extraction), and n-butanol (4 times of extraction) to obtain extractive solution. Concentrating the extractive solution under reduced pressure to obtain n-butanol extract which is total saponin enriched. Loading the n-butanol extraction part onto 300 mesh silica gel column, eluting with chloroform, chloroform-methanol (95:5), chloroform-methanol-water (7:3:0.5), methanol-water (9:1), and concentrating the chloroform-methanol-water (7:3:0.5) eluate under reduced pressure to obtain 375g of total saponins of radix Cynanchi auriculati Shemu.
Example 7
Concentrating the eluate obtained in example 1, evaporating under reduced pressure, pulverizing, sieving to obtain dry extract powder, and making into capsule.
Example 8
Concentrating the eluent obtained in example 1, evaporating to dryness under reduced pressure, pulverizing, and sieving to obtain dry extract powder. Mixing the above dry extract powder with starch and dextrin, granulating with ethanol as wetting agent, adding magnesium stearate, and making into tablet.
Experimental example 1 evaluation of Activity of Total saponins of Alternaria sinica Diels Shemu for treating ulcerative colitis caused by sodium dextran sulfate
1. Experimental materials
1.1 Main instrument
HHS-11-1 constant temperature water bath, medical equipment factory of Shanghai Boqing Utility company; 5424R small high-speed refrigerated centrifuge, ai Bende, china Co., ltd; one ten thousandth of electronic balance, BT 124S beijing certolis instruments systems limited; vortex oscillator, LAB-BLOGEN double-speed technology VTX-E; a water purifier (Milli-Q Academic A10 ultra-pure water system), millipore company, U.S.A.; ultralow temperature (-80 ℃) refrigerator, zemoer feishier technology (china) limited; an enzyme-labeled analyzer (Molecular Devices), molecular instruments, U.S.A.; ultrasonic cleaner, ningbo Xinzhi biotechnology Co., ltd.
1.2 Main medicine and kit
Sulfasalazine enteric coated tablet (0.25. 0.25 g, shanghai Yi balance pharmaceutical Co., ltd., lot: 09210601); mouse Interleukin (IL) -6ELISA kit (Hangzhou Union Biotechnology Co., ltd., lot number: A10600934), IL-1 beta ELISA kit (Hangzhou Union Biotechnology Co., ltd., lot number: A201B 11115), tumor Necrosis Factor (TNF) -alpha ELISA kit (Hangzhou Union Biotechnology Co., ltd., lot number: A28200551), myeloperoxidase (MPO) activity detection kit (Hangzhou Union Biotechnology Co., ltd., lot number: A213300713). Dextran sodium sulfate (Dextran sulfate sodium salt, DSS; MW:36000-50000;MP Biomedicals company, lot number: Q8378). Other chemical reagents were purchased from Kunming Miao technologies Inc.
1.3 Experimental animal
C57BL/6 mice, male, 20±2 g, purchased from henna schrader laboratory animals inc., license number: SCXK (Hunan) 2021-0002. Mouse feed and pads for experiments were purchased from the biological technology limited of lakeside, license number: 2016-0002. After the experimental animals entered the animal feeding house, the experimental animals were fed adaptively under laboratory conditions for 1 week. The experimental method and the conditions are inspected to be qualified by the ethical committee of animal experiment centers of the university of Chinese medicine in Yunnan, and the number of animal ethical inspection files is as follows: r-062021004.
1.4 Preparation of main reagents
3% DSS solution, weighing DSS 30 g, adding distilled water 1000 mL, and mixing well to dissolve completely. And (5) storing at 4 ℃ for standby.
Sulfasalazine suspension: taking 3 pieces of sulfasalazine enteric coated tablets (0.25 g), carefully scraping off enteric coatings by a surgical knife, fully grinding, adding 0.5% CMC-Na solution to uniformly suspend until the final volume is 25 mL, and obtaining sulfasalazine suspension (0.03 g/mL) for later use. The stomach filling volume is 0.1 mL/10 g, and the administration dosage is 0.3 g/kg.
Preparation of white dorsum Shemu total saponin sample: as shown in example 1.
White dorsum Shemu total saponin high dose group liquor: a proper amount of total saponins of white back Shemu prepared in example 1 is prepared into a solution of 10 mg/mL by distilled water. The stomach filling volume is 0.1 mL/10 g, and the administration dosage is 100 mg/kg.
White back Shemu total saponins low dose group liquor: a proper amount of total saponins of white back Shemu prepared in example 1 is prepared into a solution of 5 mg/mL by distilled water. The stomach filling volume is 0.1 mL/10 g, and the administration dosage is 50 mg/kg.
Experimental method
2.1 Animals were grouped, model and dosed randomly into normal groups, model groups, positive drug groups (sulfasalazine, 0.3 g/kg), white back Shemu total saponin high dose group (100 mg/kg), white back Shemu total saponin low dose group (50 mg/kg), 8 each. The experiment started when diary was day 0. Except for the normal group, 3% DSS solution was freely drunk from day 0 of the start of the experiment, and when the mice began to develop significant hematochezia (about 4-6 days), the DSS solution was replaced with pure water until day 10 of the experiment. The 3% DSS solution was changed every 2 days. Each group was dosed 1 time a day for 10 consecutive days starting on day 0 of the experiment. Both the normal and model groups were given equal volumes of 1% CMC-Na solution.
2.2 Mouse Disease Activity Index (DAI) score
From the day of modeling until the end of the experiment, the mice were observed daily for 10 days for mental state, feeding, hair regularity and gloss, body weight, stool characteristics and occult blood conditions, and scored according to literature standards for weight loss, stool characteristics and occult blood conditions. The weight of the animals was reduced by a percentage of the previous day (not 0, 1% -5% 1 min, 5% -10% 2 min, 10% -20% 3 min, more than 20% 4 min), stool characteristics (normal 0, diluted stool 2 min, watery stool 4 min) and stool bleeding (normal 0min, occult blood positive 2 min, dominant bleeding 4 min). The above scores are added to obtain the disease activity index (Disease activity index, DAI) of the mice.
2.3 Sample collection
On day 9 of the experiment, mice were fasted without water for 12 hours, and on day 10, 2 hours after the end of dosing, cervical vertebrae were removed and sacrificed. The mice were dissected and the entire colon was trimmed, carefully cleaned mesentery, placed neatly on filter paper, and colon length was measured and recorded. The contents of the intestinal lumen were removed by rinsing with normal saline, the middle part of the colon was carefully cut to about 0.5. 0.5 cm, and the solution was quickly rinsed with normal saline and then fixed in a 4% paraformaldehyde solution. The remaining colon was collected and immediately placed in liquid nitrogen, after the experiment was completed, transferred to a-80 ℃ refrigerator for storage.
2.4 Colonography examination
After tissue fixation is complete, it is routinely dehydrated, embedded, sectioned, and stained. The colon tissue inflammation degree, crypt injury, inflammation depth, inflammation infiltration area and lesion depth were observed under a microscope, and were scored at random double blindness.
2.5 MPO activity detection
Appropriate amounts of colon tissue were taken from each group for storage, colon tissue homogenates were prepared and the procedure was completed according to the kit instructions, and absorbance was measured for each tube at 460 nm.
MPO livingForce =
2.6 Detection of inflammatory factor content in colon tissue
2.6.1 Preparation of tissue homogenates
Shearing frozen colon tissue sample 50-mg, placing into a glass homogenizer, adding 500 μl of physiological saline, homogenizing thoroughly, and collecting tissue homogenate to obtain 10% tissue homogenate, and freezing at-20deg.C for use.
2.6.2 ELISA detection of inflammatory factors IL-1 beta, TNF-alpha and IL-6
All reagents were removed to room temperature and equilibrated for 30 minutes. Working solution, standard solution and sample diluent are respectively prepared according to the instruction of the kit, and the determination is carried out. According to the standard curve, the content of the corresponding cytokine in each sample is calculated.
2.7 Statistical analysis of data
The experimental data are analyzed and processed by adopting SPSS 21.0, and the mean value is +/-standard deviation ± the mean valueSD) represents analysis using one-way analysis of variance (ANOVA). Data were analyzed for variance alignment, with Homogeneity of Variances, LSD and Tamhane's T2, with variance alignment.
Experimental results
3.1 Effects on the Disease Activity Index (DAI) of mice
From day 0 of the experiment, each group of mice was free to drink 3% DSS solution, except for the normal group. Mice gradually develop physical signs of listlessness, hair erection, reduced activity, reduced appetite, weight loss, and the like. Beginning on day 5 of the experiment, some mice developed diarrhea, a small amount of macroscopic hematochezia, and most mice developed diarrhea and hematochezia on day 6 of the experiment. On day 6 3% DSS solution was changed to tap water for each modeling group. Model group mice developed the most severe degree of hematochezia and diarrhea by day 6; the mice in the model group have the effect of relieving the hematochezia and diarrhea from the 8 th day to the 10 th day of the experiment.
Table 1 influence of white back Shemu total saponins on DSS-induced UC model mouse DAI (n=8)
Note that: in comparison with the normal group, ## ：p<0.01; in comparison with the set of models, * ：p<0.05， ** ：p<0.01
as can be seen from Table 1, the high and low dosage groups of the total saponins of the white back Shemu and the positive medicine sulfasalazine can obviously reduce DAI scores of UC model mice. The high and low doses of the total saponins of the white back Shemu have the same effect on reducing DAI as the positive medicine sulfasalazine, but have no obvious dose dependency.
3.2 Effects on changes in colon length in mice
Table 2 influence of white dorsum Shemu total saponins on DSS-induced changes in colon length in mice with UC model (n=8)
Note that: in comparison with the normal group, ## ：p<0.01; in comparison with the set of models, * ：p<0.05
as can be seen from table 2, the colon length of the model mice is significantly shortened compared with that of the normal mice, and the high dosage of the total saponins of dorsum albuminosus Shemu can significantly improve the phenomenon.
3.3 Effects on colon histopathological changes in mice
From fig. 1 and table 3, the normal group has complete tissue structure, and the intestinal gland is abundant, and no obvious abnormality is seen. The model group has large amount of intestinal glands disappeared, replaced by the proliferated connective tissue, and is accompanied by more inflammatory cell infiltration, mainly comprising lymphocytes and neutrophils, and the inflammation infiltrates to submucosa. Compared with the model group, the aralia elata high-dose group can obviously reduce the score of colon pathological tissue slices, the slices show that the tissue structure is complete, the intestinal gland number is increased, and the aralia elata high-dose group has an improvement effect on UC colon pathological tissue lesions.
Table 3 effect of white back Shemu total saponins on DSS-induced UC model mice colon histopathological score (n=8)
3.4 Influence on the MPO Activity of colon tissue of mice
TABLE 4 influence of white dorsum Shemu Total saponins on the MPO viability of colon tissue of mice (n=8)
Note that: in comparison with the normal group, ## ：p<0.01; in comparison with the set of models, * ：p<0.05， ** ：p<0.01
as can be seen from Table 4, the model mice have higher activity of MPO than normal mice, and the high and low dosage groups of total saponins of white dorsum Shemu and the positive drug sulfasalazine can significantly improve the phenomenon.
3.5 Effect on the inflammatory factor content of the colon tissue of mice
As can be seen from tables 5,6 and 7, the amounts of inflammatory factors IL-1. Beta., TNF-. Alpha.and IL-6 in colon tissue of model mice were significantly higher than those of normal mice. The high and low doses of the total saponins of the white dorsum Shemu and the positive medicine sulfasalazine can obviously reduce the content of IL-1 beta in colon tissues of mice. The high dosage of the total saponins of the white back Shemu and the positive medicine sulfasalazine can obviously reduce the content of TNF-alpha in colon tissues of mice, wherein the high dosage of the total saponins of the white back Shemu has better effect than that of the low dosage and the positive medicine sulfasalazine. The total saponins of the white dorsum Shemu have no obvious influence on the content of IL-6 in colon tissues of mice. It is shown that the total saponins of the back Shemu can obviously reduce the inflammatory response of the colon part of the UC model mouse.
TABLE 5 influence of white dorsum Shemu Total saponins on the IL-1 beta content of inflammatory factors in colon tissue of mice (n=8)
Note that: in comparison with the normal group, ## ：p<0.01; in comparison with the set of models, * ：p<0.05， ** ：p<0.01
TABLE 6 Effect of white dorsum Shemu Total saponins on the content of inflammatory factor TNF- α in colon tissue of mice (n=8)
Note that: in comparison with the normal group, ## ：p<0.01; in comparison with the set of models, * ：p<0.05， ** ：p<0.01
TABLE 7 Effect of white dorsum Shemu Total saponins on the IL-6 content of inflammatory factors in colon tissue of mice (n=8)
Note that: in comparison with the normal group, ## ：p<0.01; comparison with model group
In conclusion, the white dorsum Shemu total saponins have remarkable activity of treating inflammatory bowel diseases, and have good application prospect in preparing medicines for treating inflammatory bowel diseases, in particular ulcerative colitis.
In the mouse test according to the invention, the total saponins of the white dorsum Shemu have a dosage of 50 mg/kg and 100 mg/kg. The doses were 300 mg and 600 mg in terms of human (calculated as adult body weight 60 kg). Considering the differences in weight of humans, and various other pathological conditions, the recommended dosage is 1-2 times daily, 150-300 mg each time. The most common dose is 2 times daily, 200 mg a time.
Claims (8)
1. The total saponins of the white back Shemu with the anti-ulcerative colitis activity are characterized in that the total saponins of the white back Shemu are total saponins of the young leaf buds of the white back Shemu, and the preparation method is realized by the following steps:
1) Pulverizing dried leaf bud of white back Shemu, extracting with methanol/ethanol, and concentrating to obtain ethanol extract;
2) Suspending the alcohol extract with water, sequentially extracting with petroleum ether, ethyl acetate and n-butanol for 2-5 times, and mixing the extracts;
3) Concentrating the n-butanol extraction part, performing silica gel column chromatography, eluting with chloroform-methanol=95:5, and sequentially eluting with chloroform: methanol: water = 7:3:0.5 elution, methanol: water = 9:1 elution, chloroform was collected: methanol: concentrating the water=7:3:0.5 part under reduced pressure to obtain the total saponins of the white back Shemu.
2. The total saponins of white back Shemu according to claim 1, wherein in step 1, the crushed leaves and buds of white back Shemu have a particle size of 20-60 mesh;
the concentration range of the ethanol is 60-85%, the concentration range of the methanol is 60-85%, and the ratio of the medicinal material amount to the methanol or ethanol is as follows: 1kg/2L to 1kg/4L, and the extraction mode is reflux extraction, decoction or leaching.
3. The total saponins of white back Shemu according to claim 1, wherein the reflux extraction temperature is 85-100 ℃, the extraction time is 30-60 min each time, and the extraction times are 2-4 times.
4. The total saponins of white back Shemu according to claim 1, wherein in step 2), when the alcohol extract is suspended in water, the ratio of the alcohol extract to water is: 1g/4 mL-1 g/10mL.
5. The total saponins of white back Shemu according to claim 1, wherein in step 2), the medium for column chromatography of n-butanol extraction site is 200-300 mesh silica gel.
6. Use of the total saponins of white back Shemu in the manufacture of a medicament for treating ulcerative colitis according to claim 1.
7. A pharmaceutical composition prepared from a therapeutically effective amount of the total saponins of quebracho Shemu of claim 1 as an active agent and a pharmaceutically effective carrier.
8. The pharmaceutical composition of claim 7, wherein the total saponins of dorsum albuminosus Shemu are dosed 1-2 times daily, 150-300 mg each time.
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