CN117942349A - Andrographolide-isatis root polysaccharide composition and preparation method and application thereof - Google Patents
Andrographolide-isatis root polysaccharide composition and preparation method and application thereof Download PDFInfo
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- andrographolide
- isatis root
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention relates to an andrographolide-isatis root polysaccharide composition and a preparation method and application thereof, which can effectively solve the problem of low bioavailability of andrographolide in treating inflammatory bowel disease.
Description
1. Technical field
The invention relates to the technical field of medicines, in particular to an andrographolide-isatis root polysaccharide composition, a preparation method and application thereof.
2. Background art
Inflammatory bowel disease is a common chronic gastrointestinal disease, which repeatedly attacks in the long-term course of the disease, is frequently found in young people, especially children, and has become a public health problem worldwide, and the pathogenesis of inflammatory bowel disease is complex, the exact mechanism is unclear, and effective therapeutic drugs are lacked. At present, the clinic is mostly treated by adopting corticosteroids, immunosuppressants or antibiotics, and the traditional Chinese medicine has poor curative effect and obvious side effect. Therefore, the development of new safe and effective therapeutic strategies has become urgent in the field of inflammatory bowel disease research.
Natural products, especially natural products derived from Chinese medicinal materials, have the advantages of multi-target synergistic effect, high safety coefficient and the like, and become the most promising sources of medicines for treating inflammatory bowel diseases. Andrographolide is a natural diterpenoid compound extracted from herba Andrographis, and is also a key pharmacodynamic material foundation of herba Andrographis for exerting heat-clearing and detoxicating, and regulating immunity. A plurality of researches prove that andrographolide can play an anti-inflammatory role through a plurality of signal paths such as NF-kappa B, JAK/STAT and the like, can effectively reduce the expression of iNOS and COX-2 and promote Th2 anti-inflammatory response. More importantly, andrographolide has excellent antibacterial activity, and can effectively control infectious inflammation by inhibiting bacterial pathogenic factors and regulating immunity. Therefore, andrographolide, also known as a "natural antibiotic", is considered as one of the most promising drugs for the treatment of inflammatory bowel disease. However, its characteristics of strong hydrophobicity and low bioavailability severely limit its clinical application.
Polysaccharide is an important natural macromolecule in Chinese medicinal materials, and has good activities of immunoregulation, antioxidation, emergency and the like. Recent studies have shown that polysaccharides can effectively alleviate intestinal inflammation by ameliorating intestinal epithelial lesions and remodelling the intestinal flora. Furthermore, the good biocompatibility and polyhydroxy structural characteristics of polysaccharides make them of great application potential in the field of drug delivery. The isatis root is a traditional Chinese medicine which is frequently matched with common andrographis herb for use, and the isatis root polysaccharide shows excellent characteristics in improving intestinal absorption of hydrophobic drugs, weakening expression of proinflammatory factors in vivo and the like, and based on network pharmacology analysis results, the isatis root polysaccharide and andrographolide have synergistic effect in anti-inflammatory aspect. However, no report on the complex use of andrographolide and isatis root polysaccharide is known at present.
3. Summary of the invention
Aiming at the situation, the invention aims to solve the defects of the prior art and provide an andrographolide-isatis root polysaccharide composition, a preparation method and application thereof, which can effectively solve the problem of low bioavailability of andrographolide in treating inflammatory bowel diseases.
The technical scheme is that the andrographolide-isatis root polysaccharide composition comprises 0.4-4 parts by weight of andrographolide and 0.5-5 parts by weight of isatis root polysaccharide, wherein the purity of the andrographolide is more than or equal to 99%, and the purity of the isatis root polysaccharide is more than or equal to 90%.
The andrographolide-isatis root polysaccharide composition is applied to the preparation of medicines for treating or improving inflammatory bowel diseases.
Further, the preparation of the medicine for treating or improving inflammatory bowel disease also comprises pharmaceutically acceptable auxiliary materials.
The andrographolide-isatis root polysaccharide composition prepared by the invention has the advantages of abundant raw material sources, simple preparation method, low preparation cost, easy mass production, good anti-inflammatory activity and high biological safety, can be used for preparing medicines for treating or improving inflammatory bowel diseases, and has practical clinical significance and popularization and application value.
4. Detailed description of the preferred embodiments
The following describes the embodiments of the present invention in further detail with reference to examples.
Example 1
In the specific implementation of the invention, the andrographolide-isatis root polysaccharide composition can also be prepared from 2-4 parts of andrographolide and 1-5 parts of isatis root polysaccharide by weight, wherein the purity of the andrographolide is more than or equal to 99%, and the purity of the isatis root polysaccharide is more than or equal to 90%.
Example 2
In the specific implementation of the invention, the andrographolide-isatis root polysaccharide composition can also be prepared from 3 parts of andrographolide and 3 parts of isatis root polysaccharide by weight, wherein the purity of the andrographolide is more than or equal to 99%, and the purity of the isatis root polysaccharide is more than or equal to 90%.
Example 3
The preparation method of the andrographolide-isatis root polysaccharide composition comprises the following steps:
1) Preparation of isatis root polysaccharide
Pulverizing radix Isatidis decoction pieces, soaking in 5 times volume of ultrapure water for 30min, decocting to boiling, decocting with slow fire for 1 hr, filtering to obtain decoction, adding 5 times volume of ultrapure water into the residue, decocting for 2 hr, filtering, mixing the two decoctions, concentrating to 500ml, adding Sevag reagent (chloroform: n-butanol=4:1, v/v), standing overnight for deproteinizing treatment, removing precipitate, repeating the steps until no precipitate is generated, concentrating the rest decoction to 500ml again, adding 4 times volume of absolute ethanol for alcohol precipitation, repeating the steps until the supernatant is colorless, filtering to remove filtrate, dissolving precipitate in hot water, adding activated carbon (feed liquid ratio 1:10, v/v) at 70deg.C for decolorizing, filtering while hot, adding 4 times of absolute ethanol into the filtrate for alcohol precipitation, filtering, washing the precipitate with absolute ethanol, acetone and diethyl ether respectively, and drying at 60deg.C to obtain radix Isatidis polysaccharide with purity not less than 90%;
2) Preparation of andrographolide-isatis root polysaccharide composition
40Ml of 10mg/ml andrographolide DMSO-water solution is dropwise added into 50ml of 2mg/ml isatis root polysaccharide solution, and stirring is carried out for 30min at room temperature, thus obtaining the product.
Example 4
Or prepared by the following steps:
1) Isatis root polysaccharide
Mixing 50ml of 10% radix Isatidis crude polysaccharide (crude polysaccharide purchased directly) solution with 50ml of 10% CTAB solution, standing at 4deg.C for 12 hr, centrifuging at 4deg.C for 20min at 3 rpm, dissolving precipitate with 30ml of 2mol/l NaCl, adding 3 times volume of 95% ethanol, mixing, standing at 4deg.C for 12 hr, centrifuging at 4deg.C for 20min at 4.C at 3 rpm at 4.0×10min, discarding supernatant, dissolving precipitate with ultrapure water, adding 1% active carbon particles, adjusting pH to 3-5, shaking at 20deg.C for 60min, centrifuging at 6.0× 10 3 rpm for 10min, removing precipitate, suction filtering supernatant, dialyzing the filtrate in dialysis bag with molecular weight cutoff of 8000-14000Da for 2 days, and lyophilizing to obtain radix Isatidis polysaccharide with purity not less than 90%;
2) Preparation of andrographolide-isatis root polysaccharide composition
40Ml of andrographolide DMSO-water solution with a concentration of 5mg/ml is dropwise added into 50ml of purified isatis root polysaccharide solution with a concentration of 2mg/ml, and the mixture is stirred for 30min at room temperature.
Example 5
Or prepared by the following steps:
1) Pulverizing radix Isatidis, adding 25 times volume of ultrapure water, decocting with slow fire for 50min, filtering, collecting supernatant, adding 1/5 times volume of chloroform and 1/25 times volume of n-butanol, shaking for 20min, centrifuging at 4.0X10 3 rpm for 20min, and lyophilizing to obtain radix Isatidis polysaccharide with purity of more than or equal to 90%;
2) Dissolving 50mg of radix Isatidis polysaccharide in ultrapure water, dropwise adding 10ml of 2mg/ml andrographolide DMSO-water solution under stirring, oscillating for 30min, centrifuging at 1.0X10 4 rpm for 30min, collecting supernatant, and centrifuging at 2.0X10 4 rpm for 60min to obtain andrographolide-radix Isatidis polysaccharide with particle diameter of 200-300 nm.
The in vitro cytological test shows that the andrographolide-isatis root polysaccharide composition not only can remarkably improve the proliferation rate of macrophages, but also can promote the polarization of M1 type macrophages with a pro-inflammatory effect into M2 type macrophages with an anti-inflammatory effect.
The colitis test of mice shows that the andrographolide-isatis root polysaccharide composition can slow down the great reduction of the weight of the mice after being orally fed to the colitis mice, effectively improve the condition of colon shortening of the colitis mice, reduce the expression of pro-inflammatory factors IL-6 and TNF-alpha cytokines in serum, and reduce the expression content of P38 alpha and COX-2 proteins in colon tissues. In addition, the andrographolide-isatis root polysaccharide composition has no obvious toxic or side effect on main organs of mice in treatment, and has good safety, and related test data are as follows:
1. evaluation of the Effect of Andrographolide-Isatis root polysaccharide composition on macrophage RAW264.7 polarization based on in vitro cell level
(1) Macrophage polarization to M1: RAW264.7 macrophages in the logarithmic growth phase are inoculated into 12-well plates, when the cell confluency reaches 70-80%, fresh culture medium is replaced, lipopolysaccharide with the final concentration of 100ng/ml is added, and the culture is continued for 24 hours to induce the macrophages to be polarized into M1 subtype.
(2) Macrophages are polarized from M1 to M2: the culture medium was discarded, replaced with fresh medium containing andrographolide-radix Isatidis polysaccharide (concentration of andrographolide and radix Isatidis polysaccharide are 20 μg/ml and 50 μg/ml respectively), and cell supernatant was collected after culturing for 24 hr, and expression levels of cytokines IL-10 and IL-12 were measured. Experimental results show that compared with a blank group, the IL-10 content of the andrographolide-isatis root polysaccharide composition treatment group is obviously increased, and the IL-12 content is obviously reduced, which indicates that the andrographolide-isatis root polysaccharide composition promotes the reversion of M1 type macrophages to M2 type macrophages.
2. Evaluation of the Effect of Andrographolide-Isatis root polysaccharide composition on proliferation of mouse Abdominal macrophages based on in vitro cell levels
(1) Isolation of mouse peritoneal macrophages: the mice were sacrificed by cervical vertebra removal after anesthesia, 4ml of sterile phosphate buffer was injected into the abdomen of the mice, gently rubbed for 2-3min, the abdominal skin was sterilized with alcohol, and then the abdominal cavity was aspirated with a syringe, centrifuged at 1.0X10 3 rpm for 5min, and the cell pellet was washed 3 times with sterile phosphate buffer. Re-suspending the cell precipitate with RPMI 1640 complete culture medium, inoculating to a 6-hole plate, culturing for 6-8h, washing off non-adherent cells with pre-warmed sterile physiological saline, and obtaining the remaining adherent cells as mouse peritoneal macrophages.
(2) Determination of proliferation rate of macrophages in abdominal cavity of mice: inoculating the mouse peritoneal macrophages into a 96-well plate, placing the mouse peritoneal macrophages into a 5% CO 2 cell incubator at 37 ℃ for incubation overnight, discarding the culture medium, and replacing the culture medium with a fresh culture medium respectively containing isatis root polysaccharide, andrographolide or andrographolide-isatis root polysaccharide composition, wherein the content of the andrographolide-isatis root polysaccharide composition is 6.25-100 mug/ml, and the mass ratio of andrographolide to isatis root polysaccharide is 2:5. After further culturing for 24 hours, the proliferation rate of the cells was measured by MTT method. The experimental results show that the proliferation rate of the macrophages in the abdominal cavity of the mice is still higher than 90 percent along with the continuous increase of the concentration of different medicines, and the cell proliferation rate of the andrographolide-isatis root polysaccharide composition group is obviously higher than that of the andrographolide-treated group.
3. Evaluation of Andrographolide-Isatis root polysaccharide effect on treatment of inflammatory bowel disease based on mouse colitis model
SPF grade 6-8 week old male C57BL/6J mice are randomly divided into 7 groups after 1 week adaptation, namely a normal control group NC, a colitis model group DSS, an andrographolide group AD, a low-dose group RIP-L/H of isatis root polysaccharide, and a low-dose group and a high-dose group AD-RIP-L/H of andrographolide-isatis root polysaccharide composition. NC groups were free to drink sterile water, the remaining groups were free to drink 3% DSS. The NC group and the DSS group are subjected to intragastric administration and are subjected to oral administration treatment by equal volumes of sterile physiological saline, and the administration doses are respectively as follows: andrographolide 100mg/kg, and radix Isatidis polysaccharide with low and high dosage of 25 and 50mg/kg respectively. The administration was carried out 1 time per day for 15 days.
The results of the treatment were evaluated at the end of the experiment using four indices, disease Activity Index (DAI), colon length in mice, TNF-alpha and IL-6 content, and are shown in Table 1. Experimental results show that compared with the NC group, the DSS group mice have different degrees of emaciation, diarrhea and hematochezia, and the DAI score is obviously increased; compared with the DSS group, the mice of each administration group have improved symptoms such as emaciation, diarrhea, hematochezia and the like, and the andrographolide-isatis root polysaccharide composition group has the best effect. After the end of the experiment, each group of mice was sacrificed from the neck, the colon was dissected and the length was measured. Experimental results showed that AD-RIP high dose mice had significantly higher colon than low dose mice and all significantly higher colon than model and other treatment groups. In addition, the expression of inflammatory factors TNF-alpha and IL-6 are also important indicators for measuring the therapeutic effect of colitis. The results show that the expression level of the cell inflammatory factors in serum of the AD-RIP composition group is obviously lower than that of the DSS group and each of the AD group and the RIP group. Experimental results show that AD-RIP treatment shows remarkable treatment effect on DSS-induced mouse colonitis, and the AD-RIP high-dose group is better than the low-dose group.
TABLE 1 Effect of different doses of Andrographolide-Isatis root polysaccharide compositions on the mice with Crohn's disease
| Index (I) | DAI relative index | Colon relative length | TNF-alpha relative expression level | IL-6 relative expression level |
| DSS | 11.80±0.45 | 5.26±0.27 | 33.52±0.04 | 45.43±0.18 |
| AD | 4.40±0.55 | 7.76±0.15 | 25.75±0.30 | 33.57±0.33 |
| RIP-L | 6.20±0.45 | 6.58±0.19 | 29.68±0.70 | 38.52±0.11 |
| RIP-H | 4.80±0.45 | 7.02±0.19 | 27.45±0.09 | 38.89±0.81 |
| AD-RIP-L | 3.40±0.55 | 8.28±0.15 | 21.53±0.51 | 19.80±0.57 |
| AD-RIP-H | 2.20±0.84 | 8.72±0.13 | 18.64±0.40 | 18.52±0.16 |
4. Evaluation of safety of Andrographolide-Isatis root polysaccharide composition Using in vivo mouse experiments
After the experiment is finished, the mice are killed by neck removal, main organs such as treat others for earnest, liver, spleen, lung, kidney and the like are solved, the mice are rinsed with normal saline, and the filter paper is weighed after being sucked dry, and the organ indexes are calculated. The calculation formula is as follows: organ index = organ weight (mg)/body weight (g).
The results are shown in Table 2, and the index of each organ of each group of mouse samples has no significant difference, which indicates that the andrographolide-isatis root polysaccharide composition does not have obvious influence on the health of mice, and the composition has good safety.
TABLE 2 Effect of different doses of Andrographolide-Isatis root polysaccharide compositions on organ index in mice with enteritis
5. Analysis of mechanism of Andrographolide-Isatis root polysaccharide composition for treating inflammatory bowel disease based on in vivo experiments in mice
After the experiment, colon tissues of each group of mice are dissected, and expression of P38 alpha and COX-2 proteins is detected by using Western Blot. Compared with NC group, DSS model group mice showed significantly increased expression level of P38 a protein in colon tissue (P < 0.0001). After treatment with the andrographolide-isatis root polysaccharide composition, the P38 alpha protein expression levels were reduced to (0.42±0.01) and (0.54±0.04) in the high-dose group and the low-dose group, respectively, as compared with the DSS group (1.02±0.02). The COX-2 protein expression level of colon tissue of each group of mice showed similar results, and the COX-2 protein expression level of andrographolide-isatis root polysaccharide composition group (high dose group 0.51.+ -. 0.02, low dose group 0.73.+ -. 0.03) was significantly lower than that of AD group (0.90.+ -. 0.06), RIP-H (1.14.+ -. 0.03), RIP-L (1.25.+ -. 0.04) and DSS group (1.42.+ -. 0.04).
6. Conclusion(s)
The invention has the advantages that the andrographolide-isatis root polysaccharide composition has satisfactory beneficial technical effects through repeated experiments, and the invention fully proves that the combination of the isatis root polysaccharide and the andrographolide overcomes the defects of andrographolide.
It is noted that the above-mentioned embodiments are merely preferred embodiments of the present invention, and the present invention is not limited thereto, and that any person skilled in the art can make modifications and equivalents within the scope of the present invention without departing from the scope of the present invention.
Claims (6)
1. An andrographolide-isatis root polysaccharide composition is characterized by comprising 0.4-4 parts by weight of andrographolide and 0.5-5 parts by weight of isatis root polysaccharide, wherein the purity of the andrographolide is more than or equal to 99%, and the purity of the isatis root polysaccharide is more than or equal to 90%.
2. The andrographolide-isatis root polysaccharide composition according to claim 1, characterized by being prepared from 2-4 parts by weight of andrographolide and 1-5 parts by weight of isatis root polysaccharide.
3. Use of an andrographolide-isatis root polysaccharide composition according to claim 1 or 2 in the preparation of a medicament for treating or ameliorating inflammatory bowel disease.
4. A method for preparing an andrographolide-isatis root polysaccharide composition according to any one of claims 1 to 3, comprising the steps of:
1) Preparation of isatis root polysaccharide
Pulverizing radix Isatidis decoction pieces, adding 5 times volume of ultrapure water, soaking for 30min, decocting to boiling, decocting with slow fire for 1 hr, filtering to obtain decoction, adding 5 times volume of ultrapure water into the residue, decocting for 2 hr, filtering, mixing the two decoctions, concentrating to 500ml, adding Sevag reagent with volume ratio of chloroform: n-butanol=4:1, standing overnight for deproteinizing treatment, removing precipitate, repeating this step until no precipitate is generated, concentrating the rest decoction to 500ml again, adding 4 times volume of absolute ethanol for alcohol precipitation, repeating this step until the supernatant is colorless, filtering to remove filtrate, dissolving precipitate in hot water, adding active carbon with feed liquid ratio of 1:10 at 70deg.C for decolorizing, filtering, adding 4 times of absolute ethanol into the filtrate for alcohol precipitation, filtering, washing precipitate with absolute ethanol, acetone and diethyl ether, and drying at 60deg.C to obtain radix Isatidis polysaccharide;
2) Preparation of andrographolide-isatis root polysaccharide composition
40Ml of 10mg/ml andrographolide DMSO-water solution is dropwise added into 50ml of 2mg/ml isatis root polysaccharide solution, and stirring is carried out for 30min at room temperature, thus obtaining the product.
5. A method for preparing an andrographolide-isatis root polysaccharide composition according to any one of claims 1 to 3, comprising the steps of:
1) Preparation of isatis root polysaccharide
Uniformly mixing 50ml of 10% radix Isatidis crude polysaccharide solution and 50ml of 10% CTAB solution, standing at 4deg.C for 12h, centrifuging at 4.0X10 3 rpm for 20min, dissolving precipitate with 30ml of 2mol/l NaCl, adding 3 times volume of 95% ethanol, mixing, standing at 4deg.C for 12h, centrifuging at 4.0X10 3 rpm at 4deg.C for 20min, discarding supernatant, dissolving precipitate with ultrapure water, adding 1% active carbon particles, regulating pH to 3-5, shaking at 20deg.C for 60min, centrifuging at 6.0X10 3 rpm for 10min, removing precipitate, vacuum filtering, dialyzing the filtrate in dialysis bag with molecular weight cutoff of 8000-14000Da for 2 days, and freeze drying to obtain radix Isatidis polysaccharide;
2) Preparation of andrographolide-isatis root polysaccharide composition
40Ml of andrographolide DMSO-water solution with a concentration of 5mg/ml is dropwise added into 50ml of purified isatis root polysaccharide solution with a concentration of 2mg/ml, and the mixture is stirred for 30min at room temperature.
6. A method for preparing an andrographolide-isatis root polysaccharide composition according to any one of claims 1 to 3, comprising the steps of:
1) Pulverizing radix Isatidis, adding 25 times volume of ultrapure water, decocting with slow fire for 50min, filtering, collecting supernatant, adding 1/5 times volume of chloroform and 1/25 times volume of n-butanol, shaking for 20min, centrifuging at 4.0X10 3 rpm for 20min, and lyophilizing to obtain radix Isatidis polysaccharide with purity of more than or equal to 90%;
2) Dissolving 50mg of radix Isatidis polysaccharide in ultrapure water, dropwise adding 10ml of 2mg/ml andrographolide DMSO-water solution under stirring, oscillating for 30min, centrifuging at 1.0X10 4 rpm for 30min, collecting supernatant, and centrifuging at 2.0X10 4 rpm for 60min to obtain andrographolide-radix Isatidis polysaccharide with particle diameter of 200-300 nm.
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