CN116492369B - Traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic effects and application thereof - Google Patents

Abstract

The invention belongs to the technical field of traditional Chinese medicines, and particularly discloses a traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic effects and application thereof. The traditional Chinese medicine polysaccharide composition comprises not less than 50% of Jiang Zhijiang elsholtzia polysaccharide by mass percent; the Chinese medicinal polysaccharide composition can remarkably reduce the bleeding time of mice and the swelling degree of feet of rats, and achieves the effects of diminishing inflammation and stopping bleeding. The traditional Chinese medicine polysaccharide composition obtained by the invention has obvious anti-inflammatory and hemostatic effects, can be applied to the preparation of oral care products, and has better anti-inflammatory and hemostatic effects if being compatible with 25% -50% of sarcandra glabra polysaccharide.

Description

Traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic effects and application thereof

Technical Field

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic effects and application thereof in preparation of oral care products.

Background

There are two types of common toothpaste and functional toothpaste on the market at present, wherein the functional toothpaste is divided into chemical toothpaste and natural herbal toothpaste. According to the related studies, it was found that the long-term use of chemical toothpaste resulted in oral problems, such as the long-term use of toothpaste with peroxide-added auxiliary whitening component resulted in easy damage of enamel and tooth sensitivity. The active ingredients of the Chinese herbal medicine added in the Chinese herbal medicine toothpaste in the domestic market are mainly flavonoids, alkaloids, saponins, volatile oils, coumarins, tannins, anthraquinones and organic acids, so that the research on oral care products with polysaccharide as the main ingredient is less, and the report of the related literature of using Jiang Zhijiang elsholtzia polysaccharide for preparing the oral care products is not seen at present. Therefore, it is of great importance to develop an oral care product which contains polysaccharide as a main ingredient and has anti-inflammatory and hemostatic effects.

Disclosure of Invention

The technical problems to be solved by the invention are as follows: aiming at the defects and deficiencies of few researches and applications of oral care products taking polysaccharide as a main component in the prior art, the invention provides a traditional Chinese medicine polysaccharide composition with anti-inflammation and hemostasis functions and application thereof in preparing the oral care products. The Chinese medicinal polysaccharide composition mainly comprises Jiang Zhijiang herba Moslae polysaccharide. In vitro cell tests prove that Jiang Zhijiang elsholtzia polysaccharide can obviously reduce the expression of NO, IL-6 and IL-12 and raise the inflammation level of IL-4 and IL-10; animal experiments also prove that the Jiang Zhijiang elsholtzia polysaccharide can obviously reduce the bleeding time of mice and the swelling degree of feet of rats; when Jiang Zhijiang, the mass ratio of the elsholtzia polysaccharide to the sarcandra glabra polysaccharide is 1: when 0.5-1.0 is combined into the traditional Chinese medicine polysaccharide composition, the traditional Chinese medicine polysaccharide composition has stronger effect on reducing the bleeding time of mice and the swelling degree of feet of rats, thereby achieving the effects of diminishing inflammation and stopping bleeding.

The invention adopts the following technical scheme to realize the aim of the invention.

A Chinese medicinal polysaccharide composition with antiinflammatory and hemostatic effects comprises at least 50% of Jiang Zhijiang herba Moslae polysaccharide by mass; the Chinese medicinal polysaccharide composition can remarkably reduce the bleeding time of mice and the swelling degree of feet of rats, and achieves the effects of diminishing inflammation and stopping bleeding.

The polysaccharide has the characteristics of green nature, small toxic and side effects, difficult drug resistance generation and the like, and is widely researched by students at home and abroad. Jiang Xiangru it is a road medicinal material of Jiangxi, contains various nutrients and active ingredients such as volatile oil, flavone, organic acid, polysaccharide and mineral elements, etc., and thus has various pharmacological effects of relieving fever, tranquilizing, relieving pain, resisting inflammation, resisting oxidation, enhancing immunity, etc., wherein the Jiangzhou Mosla polysaccharide is the active ingredient of Jiangzhou Mosla with main function. As a Chinese medicinal material of herb class, its preparation is mostly clean and cut, and "the preparation of herba Moslae with auxiliary materials" the stem removed ginger juice is stir-baked "is mentioned for the first time in Yi Zhu Ji Yi Zhu in Yi Zhong Meng. Ginger is used as a common medicinal auxiliary material, has the characteristics of relieving exterior syndrome, dispelling cold, warming middle-jiao and relieving vomiting, and can change the drug property in the process of processing different traditional Chinese medicines, thereby achieving the effects of reducing toxicity and enhancing efficiency. The invention provides the ginger processing Jiang Xiangru for the first time to extract polysaccharide and verify the anti-inflammatory and hemostatic effects of the polysaccharide.

Further, the traditional Chinese medicine polysaccharide composition also comprises 25-50% of sarcandra glabra polysaccharide by mass percent; the traditional Chinese medicine polysaccharide composition containing the ginger-processed elsholtzia polysaccharide and sarcandra glabra polysaccharide has stronger effect on reducing the bleeding time of mice and the swelling degree of feet of rats, and has more obvious anti-inflammatory and hemostatic effects.

The sarcandra glabra whole plant can be used as medicine, contains various active ingredients such as coumarin, flavonoid, phenolic acid, terpenoid, polysaccharide and the like, and is mainly used for clearing heat and cooling blood, activating blood and resolving macula, and dispelling wind and dredging collaterals. According to related reports, sarcandra glabra polysaccharide is a water-soluble component in sarcandra glabra, the sarcandra glabra polysaccharide at the isolated part of sarcandra glabra is an effective part for resisting immune thrombocytopenic purpura, and some acidic polysaccharide structures in sarcandra glabra have good anti-inflammatory and antioxidant activities.

The invention discusses the anti-inflammatory and hemostatic effects by extracting the polysaccharide from Jiang Zhijiang elsholtzia and matching with sarcandra glabra polysaccharide, has great practical significance for research and development of the polysaccharide, and also provides theoretical basis for development and utilization of oral care products.

Furthermore, the Chinese medicinal polysaccharide composition also comprises pharmaceutically acceptable auxiliary materials.

The extraction method of Jiang Zhijiang elsholtzia polysaccharide comprises the following steps: s1, preparing ginger juice; s2, processing Jiang Xiangru with ginger juice; s3 Jiang Zhijiang extraction of elsholtzia polysaccharide: the elsholtzia haichowensis which is processed by ginger juice is subjected to pretreatment, hot water extraction, reduced pressure concentration and Sevag method to remove free protein, ethanol precipitation, drying and grinding to obtain Jiang Zhijiang elsholtzia haichowensis polysaccharide.

Further, the preparation of the ginger juice in step S1 is as follows: washing fresh rhizoma Zingiberis recens with clear water, cleaning, slicing, air drying, adding distilled water after the surface of rhizoma Zingiberis recens is dried, decocting for 2-4 times, mixing filtrates, concentrating under reduced pressure to obtain rhizoma Zingiberis recens crude drug with a mass concentration of 0.8-1.2 g/mL.

Preferably, the mass volume ratio of the ginger to the distilled water is 1:3-5.

Further, in step S2, the processing Jiang Xiangru with ginger juice is as follows: putting Jiang Xiangru decoction pieces into a closed container, soaking in equal volume of rhizoma Zingiberis recens juice, and parching to obtain rhizoma Zingiberis recens juice.

Preferably, the moistening time is 4-8h, and the stir-frying time is 6-10min.

Further, the preprocessing in step S3 is as follows: adding distilled water into herba Moslae processed with rhizoma Zingiberis recens juice, and pre-treating under ultrasonic extraction condition.

Preferably, the mass volume ratio of the Jiang Zhijiang elsholtzia to the distilled water is 1:15-25, ultrasonic time is 8-12min, and ultrasonic power is 80w.

Further, the hot water leaching and the reduced pressure concentration in the step S3 are as follows: extracting pretreated herba Moslae Jiang Zhijiang with hot water, vacuum concentrating the filtrate to 0.15-0.25g/mL of crude drug.

Preferably, the hot water leaching temperature is 85-95 ℃ and the hot water leaching time is 1-2h; the reduced pressure concentration temperature is 52-60 ℃, and the reduced pressure concentration rotating speed is 40-60rpm/min.

Further, the Sevag method in step S3 removes free proteins as follows: first, according to Sevag reagent and concentrate 1:4-6, adding a Sevag reagent into the concentrated solution, shaking and centrifuging vigorously, standing and layering, centrifuging, and taking supernatant; secondly, the Sevag reagent was mixed with the supernatant at a ratio of 1:4-6, adding a Sevag reagent into the supernatant, and carrying out severe shaking centrifugation to obtain a purified supernatant; the supernatant was repeatedly treated 0-2 times under the above conditions.

Preferably, the Sevag reagent is prepared from chloroform and n-butanol in a ratio of 4:1, preparing the mixture; the centrifugal speed is 2500-3500rpm/min, and the centrifugal time is 4-8min.

Further, the ethanol precipitation in step S3 is: adding 92% -99% ethanol into the supernatant, and precipitating with ethanol overnight in a refrigerator at 4deg.C.

Preferably, the volume ratio of the supernatant to 92% -99% ethanol is 1:2.5-3.5.

Further, in step S3, the dry grinding is as follows: removing supernatant after alcohol precipitation overnight, centrifuging to collect bottom precipitate, drying, and grinding into powder to obtain rhizoma Zingiberis recens Jiang Xiangru polysaccharide.

Preferably, the centrifugation is centrifugation at 4500-5000rpm/min for 8-12min to collect the bottom precipitate, and the drying is freeze-drying.

The extraction method of sarcandra glabra polysaccharide includes the following steps:

s10, taking sarcandra glabra whole herb decoction pieces, appropriately crushing, adding distilled water, carrying out reflux extraction for a plurality of times, carrying out suction filtration, merging the extracting solutions, and carrying out reduced pressure concentration to obtain concentrated solution;

s20, adding ethanol into the concentrated solution, and placing the mixture in a refrigerator at a temperature of 4 ℃ for alcohol precipitation overnight;

s30, discarding supernatant, centrifuging the lower sediment, collecting the sediment at the bottom, freeze-drying the sediment, and grinding the sediment into powder after drying to obtain the sarcandra glabra polysaccharide.

Further, in step S10, the ratio of the sarcandra glabra whole herb decoction pieces to distilled water is 1:15-25, wherein the reflux extraction is carried out for 2-4 times, each time for 3-5 hours; the reduced pressure concentration is carried out at 52-60deg.C and 40-60 rpm/min until the mass concentration of herba Pileae Scriptae crude drug is 0.08-0.12 g/mL.

Further, in step S20, the volume ratio of ethanol added to the concentrate is: 2-4 times of 92% -99% ethanol is added into 1 volume of concentrated solution.

Further, the centrifugation in step S30 is: centrifuging at 2500-3500 rpm/min for 8-12min.

The traditional Chinese medicine polysaccharide composition has obvious anti-inflammatory and hemostatic effects, and can be applied to preparing oral care products; the oral care products include, but are not limited to, toothpastes. The invention performs experimental discussion and research and development on the anti-inflammatory and hemostatic effects of the polysaccharide before and after processing and the compatibility sarcandra glabra polysaccharide, and provides a theoretical basis for development and utilization of oral care products.

A Chinese medicinal toothpaste with antiinflammatory and hemostatic effects comprises, by mass, 0.1-1% of Chinese medicinal polysaccharide composition, 65-85% of toothpaste raw materials, and the balance of solvent; the traditional Chinese medicine polysaccharide composition comprises Jiang Zhijiang elsholtzia polysaccharide; the solvent is preferably pure water.

Further, the traditional Chinese medicine polysaccharide composition also comprises sarcandra glabra polysaccharide; the ratio of the Jiang Zhijiang elsholtzia polysaccharide to the sarcandra glabra polysaccharide is 1:0.5-1.

Further, the toothpaste raw materials include, but are not limited to, abrasives, thickeners, humectants, and foaming agents.

Furthermore, the toothpaste raw and auxiliary materials also comprise one or more of preservative, flavoring agent, essence and colorant.

Preferably, the abrasive is preferably calcium carbonate and/or silica; the thickener is preferably sodium carboxymethylcellulose (CMC-Na) and/or xanthan gum; the humectant is preferably sorbitol and/or glycerin; the foaming agent is preferably Sodium Dodecyl Sulfate (SDS), and the preservative is preferably nipagin ester; the flavoring agent is preferably saccharin sodium; the essence is preferably plant essential oil such as peppermint oil, spearmint essential oil, etc.

The beneficial effects are that:

(1) The ginger processed Jiang Xiangru polysaccharide is taken as a main component, and the obtained traditional Chinese medicine polysaccharide composition has obvious anti-inflammatory and hemostatic effects and can be applied to preparing oral care products. If the compound is combined with proper sarcandra glabra polysaccharide, the anti-inflammatory and hemostatic effects are better.

(2) According to the invention, the mouse tail-breaking hemostasis test proves that Jiang Xiangru polysaccharide has no significant difference on the bleeding time of mice, jiang Zhijiang elsholtzia polysaccharide and sarcandra glabra polysaccharide can significantly reduce the bleeding time of mice, and the bleeding time of sarcandra glabra polysaccharide in a medium-dose group of mice is significantly reduced, which is similar to the treatment effect of a Yunnan white drug powder group. The sarcandra glabra polysaccharide has better hemostatic effect. When sarcandra glabra polysaccharide and Jiang Xiangru polysaccharide or sarcandra glabra polysaccharide and Jiang Zhijiang elsholtzia polysaccharide are used respectively, the mass ratio is 0.7:1, compared with a blank control group, the sarcandra glabra polysaccharide and Jiang Zhijiang elsholtzia polysaccharide group can obviously reduce the bleeding time of mice, which shows that Jiang Zhijiang elsholtzia polysaccharide is compatible with a proper amount of sarcandra glabra polysaccharide, has a synergistic effect and better anti-inflammatory and hemostatic effects.

(3) Animal experiments prove that the elsholtzia polysaccharide, the Jiang Zhijiang elsholtzia polysaccharide and the sarcandra glabra polysaccharide can obviously reduce the foot swelling of rats, wherein Jiang Zhijiang elsholtzia polysaccharide has obvious difference in a medium-dose group, which indicates that Jiang Zhijiang elsholtzia polysaccharide can reduce the foot swelling of rats.

(4) In vitro cell experiments prove that Jiang Xiangru polysaccharide can obviously reduce pro-inflammatory factors IL-6, IL-12 and NO in a high-dose group and increase the expression of anti-inflammatory factors IL-4 and IL-10; while Jiang Zhijiang herba Moslae polysaccharide can remarkably reduce the expression of IL-6, IL-12 and NO in the medium dose group, and can increase the expression of IL-4 and IL-10 in the high dose group, which shows that the herba Moslae polysaccharide processed with ginger juice has stronger anti-inflammatory effect.

(5) The invention is proved by animal experiments of white rabbits: compared with comparative examples 1-4, the traditional Chinese medicine toothpaste (example 5) prepared from the ginger Jiang Xiangru polysaccharide and the sarcandra glabra polysaccharide can obviously improve the platelet aggregation rate and the content of platelet secretion TXB2 in fresh blood of white rabbits at the same dosage concentration of a toothpaste sample, and is also higher than that prepared from Jiang Xiangru polysaccharide prepared from ginger alone (example 4); experiments with the rat gingivitis model also demonstrated: compared with comparative examples 1-4 under the same conditions, the traditional Chinese medicine toothpaste (example 5) prepared from the ginger processed Jiang Xiangru polysaccharide and the sarcandra glabra polysaccharide can obviously reduce the levels of GI (gum index), PI (plaque index) and BOP (bleeding index), has better effects on improving anti-inflammatory factors IL-4 and IL-10 and reducing inflammatory factors such as IL-1 beta, TNF-alpha and IL-6, and has better effect than the traditional Chinese medicine toothpaste prepared from the ginger processed Jiang Xiangru polysaccharide alone (example 4). The Jiang Zhijiang elsholtzia polysaccharide is compatible with a proper amount of sarcandra glabra polysaccharide, has a synergistic effect and has better anti-inflammatory and hemostatic effects.

(6) The Jiang Zhijiang elsholtzia polysaccharide and sarcandra glabra polysaccharide preparation method provided by the invention has the advantages of unique method, simple process, convenience in operation, low cost, high polysaccharide yield and the like, and the related equipment is conventional equipment.

Drawings

Fig. 1: determining the hemostatic time of the mice;

fig. 2: measurement of rat foot swelling;

fig. 3: effects of Jiang Xiangru polysaccharide on RAW264.7 cell activity;

fig. 4: effects of Jiang Zhijiang Elsholtzia polysaccharide on RAW264.7 cell activity;

fig. 5: effect of different concentrations of LPS on RAW264.7 cell activity;

fig. 6: effect of different concentrations of LPS on the amount of NO released by RAW264.7 cells;

fig. 7: influence of polysaccharide on LPS-induced RAW264.7 cell pro-inflammatory factor NO expression;

fig. 8: effects of polysaccharide on LPS-induced RAW264.7 cell pro-inflammatory factor IL-6 expression;

fig. 9: effects of polysaccharide on LPS-induced RAW264.7 cell pro-inflammatory factor IL-12 expression;

fig. 10: effects of polysaccharide on LPS-induced expression of RAW264.7 cell anti-inflammatory factors IL-4 and IL-10;

fig. 11: influence of each toothpaste sample concentration on platelet aggregation rate in rabbit platelet biological model;

fig. 12: the concentration of each toothpaste sample can secrete TXB to the platelets in the rabbit platelet biological model ₂ Influence of content;

fig. 13: analysis of HGECs cell proliferation and toxicity for each toothpaste sample;

fig. 14: measuring cytokines IL-4 and IL-10 secreted by HGECs after LPS induced incubation of each toothpaste sample;

fig. 15: measuring cell factor NO secreted by HGECs and IL-1 beta after LPS induced incubation of each toothpaste sample;

fig. 16: measuring cell factors IL-6 and TNF-alpha secreted by HGECs after LPS induced incubation of each toothpaste sample;

fig. 17: effects of each toothpaste sample on rat oral GI (gingival index), PI (plaque index), BOP (bleeding index);

fig. 18: effects of each toothpaste sample on the content of anti-inflammatory factors (IL-4, IL-10);

fig. 19: effect of each toothpaste sample on the content of the pro-inflammatory factor IL-1 beta;

fig. 20: effect of each toothpaste sample on the amount of inflammatory factor IL-6;

fig. 21: effect of each toothpaste sample on the amount of the pro-inflammatory factor TNF-alpha.

Detailed Description

Example 1 preparation of Elsholtzia seu Moslae polysaccharide

Taking 40g Jiang Xiangru decoction pieces, performing ultrasonic assisted extraction, adding distilled water 800mL according to the mass volume ratio of 1:20, performing ultrasonic power 80w for 10min, performing reheat water extraction at 90 ℃ for 1h, performing suction filtration to obtain filtrate, performing reduced pressure concentration at 55 ℃ until the total volume is 1/4, wherein the concentration of the elsholtzia crude drug contained in the concentrate is 0.22g/mL, adding 180mL of the concentrate into 36mL of Sevag reagent with the volume of 1/5 of the concentrate, vibrating, standing and layering, centrifuging at the centrifugal speed of 3000rpm/min for 5min, taking supernatant, and repeating for three times; about 150mL of supernatant is added with 450mL of 95% ethanol with 3 times volume, and the mixture is placed in a refrigerator with 4 ℃ for alcohol precipitation overnight; the supernatant was discarded, the lower precipitate was centrifuged at 4800rpm/min for 10min to collect the bottom precipitate, and the precipitate was dried in a freeze dryer and ground to a powder after drying to give Jiang Xiangru polysaccharide 1.38g. The yield was 3.45%.

Example 2: jiang Zhijiang preparation of herba Moslae polysaccharide

Washing silt outside fresh ginger with clear water, washing, slicing, adding distilled water according to the mass-to-volume ratio of 1:4 after the surfaces of the ginger slices are dried, heating, boiling and extracting for 3 times for 30min each time, combining the filtrates, and concentrating under reduced pressure until the mass concentration of the ginger crude drug is 1g/mL to obtain ginger juice; putting Jiang Xiangru decoction pieces 40g into a No. 6 self-sealing bag, adding 40mL of equal volume of ginger juice, standing in shade, moistening for 6 hours, and parching for 8 minutes to obtain Jiang Xiangru processed ginger juice; firstly, carrying out ultrasonic auxiliary extraction, namely adding 800mL of distilled water according to the mass-volume ratio of the feed liquid of 1:20, and carrying out pretreatment Jiang Zhijiang on elsholtzia for 10 minutes under the condition of ultrasonic power of 80 w; extracting herba Moslae Jiang Zhijiang after ultrasonic treatment with hot water at 90 deg.C for 1 hr, filtering, concentrating the extractive solution at 55 deg.C at 50rpm/min until 1/4 of the total volume, and concentrating to obtain herba Moslae crude drug with mass concentration of 0.22g/mL; 180mL of the concentrated solution is added with 36mL of Sevag reagent with the volume of 1/5 of that of the concentrated solution, the concentrated solution is vibrated, is subjected to standing delamination, is centrifuged, the centrifugal speed is 3000rpm/min for 5min, and the supernatant is taken and repeated three times. About 150mL of supernatant is added with 450mL of 95% ethanol with 3 times volume, and the mixture is placed in a refrigerator with 4 ℃ for alcohol precipitation overnight; removing supernatant, centrifuging the lower precipitate for 10min at 4800rpm/min, collecting bottom precipitate, drying the precipitate in a freeze dryer, and grinding into powder to obtain rhizoma Zingiberis recens Jiang Xiangru polysaccharide 1.514g with extraction rate of 3.78%.

Example 3: preparation of sarcandra glabra polysaccharide

Taking 40g of sarcandra glabra whole herb decoction pieces, properly crushing, adding 800mL of distilled water according to a feed liquid ratio of 1:20, carrying out reflux extraction for 3 times, 4 hours each time, combining the extracting solutions, carrying out suction filtration and concentration, setting the concentration temperature to 55 ℃ in a reduced pressure concentration mode, setting the concentration rotating speed to 50rpm/min, and concentrating until the total volume is 1/4 to 1/5, thus obtaining 500mL of concentrated solution, wherein the mass concentration of the sarcandra glabra crude drug contained in the concentrated solution is 0.08g/mL; 500mL of the concentrated solution is added with 1500mL of 95% ethanol with 3 times volume, and the mixture is placed in a refrigerator with the temperature of 4 ℃ for alcohol precipitation overnight; removing supernatant, centrifuging the lower precipitate at 3000rpm/min for 10min, collecting bottom precipitate, drying the precipitate in a freeze dryer, and grinding into powder to obtain sarcandra glabra polysaccharide 2.523g with yield of 6.31%.

Example 4: preparation of toothpaste sample containing rhizoma Zingiberis recens-processed herba Moslae polysaccharide

Proportioning: the mass percentage is as follows: anti-inflammatory hemostatic Jiang Zhijiang herba Moslae polysaccharide 0.21% (corresponding to crude drug amount of 0.21%/3.78% = 0.05g crude drug/g), foaming agent SDS (sodium dodecyl sulfate) 1%, abrasive agent calcium carbonate 40% and silicon dioxide 3The weight percent of the thickener CMC-Na (sodium carboxymethylcellulose) is 0.8 percent, the weight percent of the xanthan gum is 0.4 percent, the weight percent of the flavoring saccharin sodium is 0.1 percent, the weight percent of the humectant glycerin is 3 percent, the weight percent of the sorbitol is 21 percent, the weight percent of the preservative nipagin ester is 0.2 percent, the weight percent of the essence is 1 percent, and the weight percent of the pure water is 29.3 percent.

The preparation method comprises the following steps: according to the formula proportion, firstly, raw materials such as SDS, silicon dioxide, calcium carbonate, CMC-Na and the like are weighed and ground; then, the raw materials of saccharin sodium, glycerin, sorbitol, xanthan gum, nipagin ester, essence and the like are dissolved in quantitative deionized water, particles can be basically absent after stirring for 50 min, ginger Jiang Xiangru polysaccharide can be added according to mass, stirring is continued for 30min to complete the process of preparing the paste, and the paste is formed after vacuum defoaming and then is filled in tubes.

Example 5: preparation of toothpaste sample containing rhizoma Zingiberis recens herba Moslae polysaccharide and herba Pileae Scriptae polysaccharide

Proportioning: the mass percentage is as follows: anti-inflammatory hemostatic agent Jiang Zhijiang herba Moslae polysaccharide 0.15% (corresponding to crude drug amount of 0.15%/3.78% = 0.0396g crude drug/g) +herba Pileae Scriptae polysaccharide 0.1% (corresponding to crude drug amount of 0.1%/6.31% = 0.0158g crude drug/g) (total crude drug amount of 0.05g crude drug/g), foaming agent SDS (sodium dodecyl sulfate) 1%, abrasive agent calcium carbonate 40% and silicon dioxide 3The weight percent of the thickener CMC-Na (sodium carboxymethylcellulose) is 0.8 percent, the weight percent of the xanthan gum is 0.4 percent, the weight percent of the flavoring saccharin sodium is 0.1 percent, the weight percent of the humectant glycerin is 3 percent, the weight percent of the sorbitol is 21 percent, the weight percent of the preservative nipagin ester is 0.2 percent, the weight percent of the essence is 1 percent, and the weight percent of the pure water is 29.3 percent.

The preparation method comprises the following steps: according to the formula proportion, firstly, raw materials such as SDS, silicon dioxide, calcium carbonate, CMC-Na and the like are weighed and ground; then, the raw materials of saccharin sodium, glycerin, sorbitol, xanthan gum, nipagin ester, essence and the like are dissolved in quantitative deionized water, particles can be basically absent after stirring for 50 min, and the elsholtzia polysaccharide and sarcandra glabra polysaccharide prepared from ginger can be added according to mass, and stirring is continued for 30min to complete the paste preparation process, and then the paste is filled in tubes after vacuum defoaming molding.

Comparative example 1: preparation of JiangxiangMosla toothpaste

Preparation of a elsholtzia composition: 500g of elsholtzia, 400g of astragalus, 300g of chrysanthemum and 150g of honeysuckle, and firstly, crushing the traditional Chinese medicine raw materials (dry products); secondly, uniformly mixing the crushed traditional Chinese medicine raw materials and adding water into the mixture to soak the mixture for 1 h; decocting the soaked Chinese medicinal materials with water for 3 times for 1 hr, 0.5 hr and 0.5 hr respectively, and filtering to obtain decoction; then combining the decoction filtrates, concentrating under vacuum of < -0.08MPa, and stopping concentrating until the medicinal liquid contains crude drug 10 g/mL to obtain the additive of Chinese herbal toothpaste.

The toothpaste comprises the following components: the mass percentage is as follows: 0.5% of elsholtzia composition (the concentration of the composition is 0.1mL/g,0.5% corresponds to the crude drug content of 0.05 g crude drug/g), 1% of foaming agent SDS (sodium dodecyl sulfate), 40% of abrasive agent calcium carbonate and 3% of silicon dioxide, 0.8% of thickening agent CMC-Na (sodium carboxymethyl cellulose) and 0.4% of xanthan gum, 0.1% of flavoring agent saccharin sodium, 3% of humectant glycerin and 21% of sorbitol, 0.2% of preservative nipagin ester, 1% of essence and 29% of pure water.

The preparation method comprises the following steps: according to the formula proportion, firstly, raw materials such as SDS, silicon dioxide, calcium carbonate, CMC-Na and the like are weighed and ground; then, the raw materials of saccharin sodium, glycerin, sorbitol, xanthan gum, nipagin ester, essence and the like are dissolved in quantitative deionized water, particles can be basically absent after stirring for 50 min, the elsholtzia composition can be added according to mass, stirring is continued for 30min to complete the process of preparing the ointment, and the ointment is formed after vacuum defogging and then is filled into tubes.

Comparative example 2: yunnan white drug powder toothpaste

Commercially available. Yunnan white drug powder group Co., ltd., 185 g, 20201222.

Comparative example 3: sarcandra glabra toothpaste

Commercially available. Nanchang Chengzhi Co., ltd., 228 g, 20201125.

Comparative example 4: yunnan pseudo-ginseng traditional Chinese medicine toothpaste

Commercially available. Buddha Huang Qijia pure bioengineering Co., ltd., 180 g, 20201015.

Experimental example 1: determination of hemostatic time in mice

130 healthy male Kunming mice, weighing 20+ -20 g. The feed is fed into an SPF-level barrier system, and the ambient temperature is 20-23 ℃ and the humidity is 45-55%. The mice were randomly divided into 13 groups (see FIG. 1 for grouping), each group was 10, respectively, normal control group (NS group), positive group (Yunnan white drug powder group 50mg/kg, YNBY group), low, medium and high dose group (35, 70, 140mg/kg, i.e., JXR-L, JXR-M, JXR-H group), jiang Zhijiang low, medium and high dose group (35, 70, 140mg/kg, i.e., JZJXR-L, JZJXR-M, JZJXR-H group), sarcandra low, medium and high dose group (35, 70, 140mg/kg, i.e., CSH-L, CSH-M, CSH-H group), sarcandra glabra polysaccharide+ Jiang Xiangru polysaccharide compatible group (25 mg/kg+35mg/kg, CSH+JXR group) and sarcandra glabra polysaccharide+ Jiang Zhijiang. Normal control group is filled with normal saline with the same volume, and the administration group is respectively filled with polysaccharide solutions with different concentrations, 1 time per day, and 6 days continuously. After the 6 th administration for 30min, cutting off the part 0.3cm away from the tip of the tail of the mouse, and when the blood overflows automatically, starting timing, sucking blood drops once every 15s by using filter paper until the filter paper is free of blood when sucking, namely the bleeding time. The measurement of hemostasis time for each group of mice is shown in figure 1. In the figure: * Indicating significant differences (P < 0.05) compared to the normal control (NS) group, and indicating significant differences (P < 0.01) compared to the normal control (NS) group, as follows.

As can be seen from an analysis of fig. 1: after different concentrations of elsholtzia polysaccharide and Jiang Zhijiang elsholtzia polysaccharide are respectively administered by stomach infusion, the bleeding time of mice is reduced to different degrees along with the increase of the administration concentration, wherein compared with a normal control group, the Jiang Xiangru polysaccharide high-dose group and Jiang Zhijiang elsholtzia polysaccharide have significant differences (P <0.05 or P < 0.01) between a medium-dose group and a high-dose group, which indicates that the hemostatic effect of Jiang Zhijiang elsholtzia polysaccharide after ginger processing is superior to Jiang Xiangru polysaccharide.

When different concentrations of sarcandra glabra polysaccharide are administered by gastric lavage, the bleeding time of mice is obviously reduced, wherein, compared with the normal control group, the sarcandra glabra polysaccharide has obvious difference (P < 0.05) in the medium dose group, and the bleeding time is obviously reduced (P < 0.01) in the high dose group. Compared with the positive group, the sarcandra glabra high dose group has no statistical significance, but has lower bleeding time for mice than the positive group. The polysaccharides of the compatibility group have no statistical significance compared with the normal control group, and the sarcandra glabra and the elsholtzia haichowensis have significant differences (P < 0.01). This shows that the compatibility of sarcandra glabra polysaccharide and Jiang Zhijiang elsholtzia polysaccharide with low dosage has obvious hemostatic effect.

Experimental example 2: measurement of rat foot swelling

112 healthy male SD rats with weight of 200+ -20 g are bred in SPF-class barrier system, and the ambient temperature is 20-23 ℃ and the humidity is 45% -55%. Rats were randomly grouped into 14 groups (see figure 2), each group being 8, respectively, normal group (NS group), model group (Model group), positive group (Aspirin group), dosing group (low, medium and high dose group of elsholtzia polysaccharide, jiang Zhijiang elsholtzia polysaccharide and sarcandra glabra polysaccharide), and compatibility group (sarcandra glabra polysaccharide+ Jiang Xiangru polysaccharide, sarcandra glabra polysaccharide+ Jiang Zhijiang elsholtzia polysaccharide). The normal group and the model group irrigate the same volume of physiological saline; the dosage of the positive group is 50mg/kg; the low, medium and high doses of the administration group are respectively 50mg/kg, 100mg/kg and 200mg/kg, the administration dose of the compatibility group is 25mg/kg+35mg/kg, and the administration is performed once daily by gastric lavage and is performed for six days. On day seven, the left hind foot of each rat was marked and the initial volume of the rat before molding was measured. Performing gastric lavage according to the experimental group, and recording gastric lavage time; after 1 hour of gastric lavage, 0.1mL of 1% carrageenan was subcutaneously infused into the middle region of the left hind paw of rats in model group, positive drug, dosing group, and compatibility group, respectively. Toe volumes were measured after injection of carrageenan for 0h, 1h, 2h, 4h, respectively. The foot swelling degree is the toe volume after carrageenan injection minus the initial volume before molding. The measurement of foot swelling in each group of rats is shown in fig. 2.

As can be seen from an analysis of fig. 2: after carrageenan injection, the toe volumes were measured after 0, 1, 2, 4h, respectively. Wherein at 0, 1 and 2 hours, the toe volumes of rats in the dosing group were all increased to different extents, but at 4 hours, the positive group was significantly different (P < 0.01) compared to the model group; the elsholtzia polysaccharide and sarcandra glabra polysaccharide have statistical significance in the high dose group (P < 0.05), while Jiang Zhijiang elsholtzia polysaccharide has significant difference in the medium dose group (P < 0.05), which indicates that Jiang Zhijiang elsholtzia polysaccharide can reduce foot swelling in rats.

When the low dose sarcandra glabra polysaccharide was combined with the low dose of elsholtzia haichowensis polysaccharide and Jiang Zhijiang elsholtzia haichowensis polysaccharide respectively, there was a significant difference (P < 0.05) in csh+jjjxr polysaccharide at 4h of carrageenan injection compared to model group, whereas csh+jxr polysaccharide did not. This demonstrates that low doses of sarcandra glabra polysaccharide and Jiang Zhijiang elsholtzia polysaccharide reduce foot swelling in rats.

Experimental example 3

(1) Effect of elsholtzia polysaccharide and Jiang Zhijiang on RAW264.7 cell activity:

RAW264.7 cells were cultured in high sugar medium containing 10% fetal bovine serum, and when the cells were grown to log phase, the cell concentration was adjusted to 2X 10 ⁵ Inoculating in 96-well plate at 37 deg.C and 200 μl per well, and CO ₂ Culturing in an incubator with the concentration of 5% for 24 hours, replacing culture mediums containing elsholtzia polysaccharide and Jiang Zhijiang elsholtzia polysaccharide with different concentrations to obtain final concentrations of 2, 5, 10, 20, 40, 60, 80 and 100 mug/mL, and arranging six compound holes at each concentration to serve as a medicine group; simultaneously, a culture medium starvation culture medium without 1% of the medicine is used as a control group; starvation medium without cells was cultured as a blank. After incubation for 24 hours, the broth was discarded, the incubation was stopped after adding 10% MTT in each well for 4 hours, the medium in the well was carefully aspirated, 150. Mu.l DMSO was added to each well, and the wells were gently shaken on a shaker for 10min, and absorbance at 490nm was measured on an microplate reader. FIG. 3 is a graph showing the effect of Jiang Xiangru polysaccharide on RAW264.7 cell activity; FIG. 4 shows the effect of ginger processed Jiang Xiangru polysaccharide on RAW264.7 cell activity.

As can be seen from an analysis of fig. 3 and 4: the method comprises the steps of interfering RAW264.7 cells with elsholtzia polysaccharide and Jiang Zhijiang elsholtzia polysaccharide with different concentrations, screening the interfering concentrations, and detecting the influence of the elsholtzia polysaccharide and Jiang Zhijiang elsholtzia polysaccharide with different mass concentrations on the proliferation rate of the cells by an MTT method. As a result, it was found that when both polysaccharides were below 10 μg/mL, there was no toxic effect on cells, and that further increasing the concentration of polysaccharides significantly inhibited cell proliferation (P < 0.05) and was dose dependent, compared to the control group.

(2) MTT assay LPS (lipopolysaccharide) -induced effect of RAW264.7 on cell activity:

adjusting the cell concentration to 2 x 10 ⁵ Inoculating 200 mu l of each well in a 96-well plate at 37 degrees and CO ₂ Culturing in 5% incubator for 24 hr, and changing LPS culture medium with different concentrations to final concentrations of 0.1, 0.2, 0.5, 1, 2 μg/mLSix compound holes are arranged under the concentration and are used as a drug administration group; simultaneously, setting a culture medium without the medicine as a control group; cell-free medium served as a blank. After 24h incubation, the broth was discarded, 10% MTT medium was added to each well, incubation was continued for 4h, the incubation was terminated, medium in the wells was carefully aspirated, 150 μl DMSO was added to each well, and the wells were gently shaken on a shaker for 10min, and absorbance at 490nm was measured on an microplate reader. FIG. 5 shows the effect of different concentrations of LPS on RAW264.7 cell activity.

As can be seen from an analysis of fig. 5: after the cells are treated by lipopolysaccharide with different concentrations, the MTT method is used for detecting the cell activity of LPS on RAW264.7 cells, compared with a normal control group, the LPS has obvious difference (P < 0.05) when the concentration is 2 mug/mL, and has an influence on the activity of the cells. The result shows that when the concentration of the LPS is 0-1 mug/mL, the cell activity is not influenced, and a reference can be provided for the selection of the LPS concentration.

(3) Effect of LPS (lipopolysaccharide) on the NO content of RAW264.7 cells:

cells were cultured as described above, and after 24 hours of incubation, cell supernatants were collected and assayed for NO content by NO kit procedure. And (3) diluting the standard substance to be 0, 1, 2, 5, 10, 20, 40, 60 and 100 mu M, and preparing a standard curve. 50 μl of sample was added to each well, then Griess1 and Griess2 were added, respectively, absorbance at 450nm was measured on a microplate reader and NO content was calculated. FIG. 6 shows the effect of different concentrations of LPS on the NO release of RAW264.7 cells.

As can be seen from an analysis of fig. 6: the influence on the release amount of NO after the RAW264.7 macrophages are induced by LPS with different concentrations is detected by using the NO kit, the content of NO is positively correlated with the concentration of the LPS along with the increase of the concentration of the LPS, and compared with a normal group, 0.1-2 mug/mL of LPS has obvious difference (P < 0.01). This indicates that LPS induces inflammation at 0.1-2 μg/mL. In combination with a cell activity test, the concentration of LPS is selected to be 1 mug/mL, and the LPS can be used for subsequent cell inflammation modeling.

(4) Detection of LPS-induced RAW264.7 cell inflammatory factors by elsholtzia polysaccharide and Jiang Zhijiang elsholtzia polysaccharide:

cells were cultured as described above, inoculated into 96-well plates, incubated for 24 hours, carefully aspirated off the medium, and washed twice with PBS. Polysaccharide solutions (2 mug/mL, 5 mug/mL and 10 mug/mL) with different concentrations are respectively dosed, and after 1h of dosing, 1 mug/mL of LPS is dosed again. LPS groups and normal control groups were set simultaneously, with 3 duplicate wells per group. After 24h incubation, cell supernatants were collected and assayed for inflammatory factors NO, IL-6, IL-12, IL-4 and IL-10 by ELISA kit. FIG. 7 is a graph showing the effect of polysaccharide on LPS-induced expression of RAW264.7 cell pro-inflammatory factor NO; FIG. 8 is a graph showing the effect of polysaccharide on LPS-induced RAW264.7 cell pro-inflammatory IL-6 expression; FIG. 9 is a graph showing the effect of polysaccharide on LPS-induced RAW264.7 cell pro-inflammatory IL-12 expression; FIG. 10 is a graph showing the effect of polysaccharide on LPS-induced expression of RAW264.7 cell anti-inflammatory factors IL-4 and IL-10.

As can be seen from the analysis of fig. 7, 8, 9 and 10: compared with a blank control group, the levels of NO, IL-6 and IL-12 in the model control group are obviously increased, the levels of IL-4 and IL-10 are obviously reduced (P <0.05 or P < 0.01), compared with the model control group, the levels of NO, IL-6 and IL-12 are respectively reduced to different degrees at medium and high doses of elsholtzia polysaccharide and Jiang Zhijiang elsholtzia polysaccharide, the levels of IL-4 and IL-10 are obviously increased (P <0.05 or P < 0.01), wherein, compared with the elsholtzia polysaccharide group, jiang Zhijiang has NO obvious difference, but at high doses, the levels of pro-inflammatory factors NO, IL-6 and IL-12 are lower than those of the elsholtzia polysaccharide, and the anti-inflammatory factors are opposite, which indicates that the processed elsholtzia polysaccharide has better anti-inflammatory effect.

Experimental example 4: six toothpaste anti-inflammatory hemostatic effects experimental comparisons

Three kinds of toothpastes of example 4 (containing ginger Jiang Xiangru polysaccharide), example 5 (containing ginger-made elsholtzia polysaccharide and sarcandra glabra polysaccharide), comparative example 1 (containing elsholtzia composition), and six kinds of toothpastes of comparative example 2 (Yunnan white drug powder toothpaste), comparative example 3 (sarcandra glabra toothpaste), comparative example 4 (Yunnan pseudo-ginseng traditional Chinese medicine toothpaste), were leached in a water bath, the leaching solution was filtered, the supernatant was centrifugally taken out, and then decompressed and spin-evaporated (40 ℃ at 75 rpm/min) to the original toothpaste volume, and dried in a vacuum drying oven for 6 hours, and then the sample for the toothpaste was obtained by freeze-drying, and was re-dissolved into a filter membrane of 0.8%,0.4%,0.2%,0.1%,0.05%,0.22 μm, and the filter membrane was filtered at 4 ℃ for later use, and in vivo animal experiments and in vitro experiments were performed under the same conditions, and the anti-inflammatory and hemostatic effects were compared.

Experimental protocol design and outcome analysis:

(1) By studying the platelet aggregation rate and thromboxane B2 (TXB ₂ ) Content index, comprehensively evaluating the effect of toothpaste on stopping bleeding of gum:

fresh blood of white rabbit, and platelet density is adjusted to 1×10 after centrifugation ¹² L, absorbance OD at the time of first detecting blank ₀ Different toothpaste samples (5 uL/well) were added and gently shaken. The 96-well plate was incubated at 37℃for 5 minutes to allow full reaction. The turbidity in the system was changed after the subsequent addition of 5uL of adenosine diphosphate ADP solution with gentle shaking to induce platelet aggregation, which occurs in the aggregated reaction. The change of OD value detected by the enzyme labeling method is used for calculating the platelet aggregation rate.

The platelet aggregation rate is the earliest reaction to the hemostatic function, and the blood coagulation state is shown when the platelet aggregation rate is increased, and fig. 11 shows the influence of each toothpaste sample concentration on the platelet aggregation rate in the rabbit platelet biological model. As can be seen from fig. 11, the platelet aggregation rate increased with increasing toothpaste concentration, and example 5 was significantly different from the other groups at each concentration (P <0.05, x).

Platelet secretion TXB ₂ The content of (2) can reflect the activity of the platelet, TXB ₂ Higher content indicates more active platelet physiological activity, FIG. 12 shows the concentration of each toothpaste sample to platelet-excrete TXB in rabbit platelet biological model ₂ Influence of the content. From fig. 12, it can be seen that example 5 significantly different from the other groups in the concentration range of 0.1% -0.4% (P<0.05, ×); example 5 and example 4 differ significantly from the other groups at a concentration of 0.8% (P<0.05, × represents).

(2) CCK-8 method to detect proliferation and toxicity of HGECs cells, and screening the optimal toothpaste sample concentration value of HGECs according to the survival rate of cells after drug addition:

FIG. 13 shows the analysis of HGECs cell proliferation and toxicity by each toothpaste sample, it is clear from FIG. 13 that the concentration gradient of toothpaste sample treatment (10. Mu.g/mL LPS is contained), CCK-8 reagent is added after full incubation, absorbance of each well at 550nm is measured by enzyme-labeled instrument, the toothpaste sample concentration with cell viability of 80% -100% is the most safe and optimal treatment concentration for HGECs, the toothpaste sample with the concentration is selected, the Lipopolysaccharide (LPS) induced inflammation model of gingival epithelial cells HEGCs (LPS induced model group) is used, the normal control group of gingival epithelial cells HEGCs is used, and the contents of cytokines NO, IL-4, IL-10, IL-6, IL-1 beta and TNF-alpha are detected to evaluate the effect in anti-inflammatory aspect.

Toothpaste samples were tested for HGECs cell proliferation and toxicity by CCK-8, and the results showed that the anti-inflammatory effect was optimal for each group of toothpaste samples using a sample concentration of 0.6 ug/mL.

The cytokines NO, IL-4, IL-10, IL-6, IL-1β, TNF- α secreted by HGECs after incubation of each toothpaste sample induced by LPS, FIG. 14 is a measurement of the cytokines IL-4, IL-10 secreted by HGECs after incubation of each toothpaste sample induced by LPS; FIG. 15 shows the measurement of secreted cytokines NO, IL-1. Beta. By HGECs after LPS-induced incubation of various toothpaste samples; FIG. 16 shows the measurement of secreted cytokines IL-6, TNF- α by HGECs after LPS-induced incubation of various toothpaste samples. As can be seen from fig. 14, 15, 16: each toothpaste sample had an increased content of anti-inflammatory factors (P < 0.05 or P < 0.01) and a decreased content of inflammatory factors (P < 0.05 or P < 0.01) compared to LPS-induced model groups, but example 5 had a better effect on the elevation of anti-inflammatory factors IL-4 and the reduction of IL-1β, TNF- α, NO (P < 0.05). In fig. 14, 15, 16, significant differences (P < 0.05) compared to the LPS-induced model group are shown, significant differences (P < 0.01) compared to the LPS-induced model group are shown, and significant differences (P < 0.05) compared to the comparative example group are shown.

(3) Through a rat gingivitis model, a probe detects a bleeding index (BOP), a Gingival Index (GI) and a bacterial Plaque Index (PI), and the contents of cytokines NO, IL-4, IL-10, IL-6, IL-1 beta, TNF-alpha and the like in rat plasma are detected to evaluate the in-vivo experimental effect of the toothpaste:

Through the gingivitis model of rats, 56 SPF SD rats, female, with a weight of 200+ -20 g, have good activity, no caries and no periodontal disease. After the rat is anesthetized by intraperitoneal injection of pentobarbital sodium, the second molar of the two sides is selected to be ligated with a medical thread at the dental neck, the ligature thread is put into the gingival sulcus as much as possible, 10% syrup is used for replacing drinking water for feeding after ligation, 0.5mL of prepared bacterial liquid is inoculated once every other day, the normal control group is a sham operation group, and 10% syrup is not fed after anesthesia without thread ligation. After modeling, the model rats were randomly divided into 7 groups, 6 groups were given samples 30d according to the experimental protocol, and the other model group and the normal control group (sham operation group) were given only physiological saline. Fixing the rat after anesthesia by injecting sodium pentobarbital into the abdominal cavity, detecting the contents of cytokines such as NO, IL-4, IL-10, IL-6, IL-1 beta and TNF-alpha in the plasma of the rat, and evaluating the in-vivo experimental effect of the toothpaste.

The gingival index GI evaluates the health of the oral cavity by checking the change of the color and quality of the gingiva, the plaque index PI evaluates the prevention and treatment effect of periodontal disease in the oral cavity according to the thickness of dental plaque, and the gingival bleeding index BOP evaluates the pathological condition of the gingiva of the oral cavity by stimulating the gingival part through a probe. (control group is sham operated normal rats, model mice are gingivitis rats).

The results showed a significant reduction in each toothpaste to the oral cavity GI, PI, BOP of rats (P < 0.05) compared to the gingivitis model of rats, wherein example 5 significantly reduced GI, PI, BOP levels (P < 0.05) compared to the comparative examples, and figure 17 shows the effect of each toothpaste sample on oral GI (gingival index), PI (plaque index), BOP (bleeding index) of rats. The change of inflammatory factors in serum can be seen that each toothpaste sample can increase the content of the inflammatory factors (P < 0.05) compared with the model group, and the content of the inflammatory factors is reduced (P < 0.05 or P < 0.01), but the embodiment 5 has better effects (P < 0.05) on the improvement of the anti-inflammatory factors IL-4 and IL-10 and the reduction of IL-1 beta, TNF-alpha and IL-6, and the effect of each toothpaste sample on the content of the inflammatory factors (IL-4 and IL-10) is shown in figure 18; FIG. 19 is a graph showing the effect of each toothpaste sample on the amount of the inflammatory factor IL-1. Beta.; FIG. 20 effect of each toothpaste sample on the amount of inflammatory factor IL-6; FIG. 21 is a graph showing the effect of each toothpaste sample on the amount of the pro-inflammatory factor TNF- α. In fig. 18, 19, 20, and 21, significant differences (P < 0.05) from the model group, significant differences (P < 0.01) from the model group, and significant differences (P < 0.05) from the comparative example group are shown.

The experimental results are analyzed and compared to show that: through the animal experiments, the comprehensive ordering of the anti-inflammatory and hemostatic effects of the six kinds of toothpaste can be obtained as follows: example 5 (containing ginger processed elsholtzia polysaccharide and sarcandra glabra polysaccharide) > example 4 (containing ginger processed Jiang Xiangru polysaccharide) > comparative example 1 (containing elsholtzia composition) > comparative example 2 (yunnan white drug paste) > comparative example 3 (sarcandra glabra paste) > comparative example 4 (yunnan pseudo-ginseng traditional Chinese medicine paste).

Wherein: compared with comparative examples 1-4, the traditional Chinese medicine toothpaste (containing the elsholtzia polysaccharide prepared by ginger and the sarcandra glabra polysaccharide) prepared in the example 5 can obviously improve the platelet aggregation rate and the platelet secretion TXB2 content in fresh blood of white rabbits at the same dosage concentration of a toothpaste sample, and is also higher than the traditional Chinese medicine toothpaste (containing the Jiang Xiangru polysaccharide prepared by ginger) prepared in the example 4, which shows that Jiang Zhijiang elsholtzia polysaccharide is compatible with a proper amount of sarcandra glabra polysaccharide, has a synergistic effect and better anti-inflammatory and hemostatic effects.

Experiments with the rat gingivitis model also demonstrated: compared with comparative examples 1-4 under the same conditions, the traditional Chinese medicine toothpaste prepared in the embodiment 5 of the invention can obviously reduce the levels of GI (gum index), PI (plaque index) and BOP (bleeding index), has better effects on improving anti-inflammatory factors IL-4 and IL-10 and reducing inflammatory factors such as IL-1 beta, TNF-alpha and IL-6, has better effect than the traditional Chinese medicine toothpaste prepared in the embodiment 4, shows that Jiang Zhijiang elsholtzia polysaccharide is compatible with a proper amount of sarcandra glabra polysaccharide, has synergistic effect and better anti-inflammatory and hemostatic effects.

The above description of the specific embodiments of the present invention has been given by way of example only, and the present invention is not limited to the above description of the specific embodiments. Any equivalent modifications and substitutions for the present invention will occur to those skilled in the art, and are also within the scope of the present invention. Accordingly, equivalent changes and modifications are intended to be included within the scope of the present invention without departing from the spirit and scope thereof.

Claims (9)

1. A Chinese medicinal polysaccharide composition with anti-inflammatory and hemostatic effects is characterized in that: comprises not less than 50% of Jiang Zhijiang elsholtzia polysaccharide and 25% -50% of sarcandra glabra polysaccharide by mass percent; the Chinese medicinal polysaccharide composition can reduce the bleeding time of mice and the swelling degree of feet of rats, and has antiinflammatory and hemostatic effects.

2. The traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic functions according to claim 1, wherein the composition is characterized in that: the extraction method of Jiang Zhijiang elsholtzia polysaccharide comprises the following steps:

s1, preparing ginger juice;

s2, processing Jiang Xiangru with ginger juice;

s3 Jiang Zhijiang extraction of elsholtzia polysaccharide: the elsholtzia haichowensis which is processed by ginger juice is subjected to pretreatment, hot water extraction, reduced pressure concentration and Sevag method to remove free protein, ethanol precipitation, drying and grinding to obtain Jiang Zhijiang elsholtzia haichowensis polysaccharide.

3. The traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic functions according to claim 1, wherein the composition is characterized in that: the extraction method of sarcandra glabra polysaccharide comprises the following steps:

s10, taking sarcandra glabra whole herb decoction pieces, crushing, adding distilled water, carrying out reflux extraction for a plurality of times, carrying out suction filtration, merging the extracting solutions, and carrying out reduced pressure concentration to obtain concentrated solution;

s20, adding ethanol into the concentrated solution, and placing the mixture in a refrigerator at the temperature of 4 ℃ for alcohol precipitation overnight;

s30, discarding supernatant, centrifuging the lower sediment, collecting the sediment at the bottom, freeze-drying the sediment, and grinding the sediment into powder after drying to obtain the sarcandra glabra polysaccharide.

4. The traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic effects according to claim 2, wherein the preparation of the ginger juice in step S1 is as follows: washing fresh rhizoma Zingiberis recens with clear water, cleaning, slicing, air drying, adding distilled water after the surface of rhizoma Zingiberis recens is dried, heating and decocting for 2-4 times, mixing filtrates, concentrating under reduced pressure to obtain crude drug with a mass concentration of 0.9-1.1g/mL to obtain rhizoma Zingiberis recens juice; the mass volume ratio of the ginger to the distilled water is 1:3-5.

5. the traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic functions according to claim 2, wherein the processing Jiang Xiangru with ginger juice in step S2 is: putting Jiang Xiangru decoction pieces into a closed container, soaking in equal volume of rhizoma Zingiberis recens juice in shade, parching to obtain Jiang Xiangru; the moistening time is 4-8 hours; the parching time is 6-10min.

6. The traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic functions according to claim 2, wherein the pretreatment in step S3 is: adding distilled water into herba Moslae processed with rhizoma Zingiberis recens juice, and pre-treating under ultrasonic extraction condition; the mass volume ratio of the Jiang Zhijiang elsholtzia to the distilled water is 1:15-25; the ultrasonic time of ultrasonic extraction is 8-12min, and the ultrasonic power is 80w; the hot water leaching and the reduced pressure concentration in the step S3 are as follows: extracting pretreated Jiang Zhijiang herba Moslae with hot water, vacuum concentrating the filtrate to crude drug mass concentration of 0.15-0.25g/mL; the hot water leaching temperature is 85-95 ℃, and the hot water leaching time is 1-2h; the reduced pressure concentration temperature is 52-60 ℃, and the reduced pressure concentration rotating speed is 40-60rpm/min.

7. The traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic functions according to claim 2, wherein the removal of free proteins by the Sevag method in step S3 is as follows: first, according to Sevag reagent and concentrate 1:4-6, adding a Sevag reagent into the concentrated solution, vibrating and centrifuging, standing and layering, centrifuging, and taking supernatant; secondly, the Sevag reagent was mixed with the supernatant at a ratio of 1:4-6, adding a Sevag reagent into the supernatant, vibrating and centrifuging to obtain a purified supernatant; repeating the treatment of the supernatant for 0-2 times; the Sevag reagent comprises chloroform and n-butanol with the ratio of 4:1, preparing the mixture; the centrifugal speed is 2500-3500rpm/min, and the centrifugal time is 4-8min.

8. The traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic functions according to claim 2, wherein the ethanol precipitation in step S3 is: adding 92% -99% ethanol into the supernatant, and precipitating with ethanol in a refrigerator at 4deg.C overnight; the volume ratio of the supernatant to 92% -99% ethanol is 1:2.5-3.5; the dry grinding in step S3 is as follows: removing supernatant after alcohol precipitation overnight, centrifuging to collect bottom precipitate, drying, and grinding into powder to obtain rhizoma Zingiberis recens Jiang Xiangru polysaccharide; centrifuging for 8-12min at 4500-5000rpm/min, and collecting bottom precipitate; the drying is freeze drying.

9. Use of a traditional Chinese medicine polysaccharide composition with anti-inflammatory and hemostatic properties according to any one of claims 1-8 in the preparation of an oral care product.