External preparation for promoting wound healing and preparation method thereof

Abstract

The invention discloses an external preparation for promoting wound healing and a preparation method thereof, and belongs to the technical field of wound treatment. The external preparation for promoting wound healing of the invention comprises: a radix scutellariae extract, a radix notoginseng extract, a radix peony root extract and borneol. The radix scutellariae extract of the invention stops bleeding and stops pain, and is the main drug; the radix notoginseng extract removes blood stasis and stops bleeding, promotes blood circulation and stops pain, and improves the hemostatic and analgesic effect of the radix scutellariae extract, and is the minister drug; the radix peony root extract clears heat and cools blood, disperses blood stasis and stops pain, and is the adjuvant drug; the borneol dredges meridians, and is the guiding drug; the drugs are used together to stop blood toxicity, reduce inflammation and relieve pain, and promote blood circulation and promote tissue regeneration. The components of the external preparation of the invention have synergistic effects, have an inhibitory effect on Staphylococcus aureus, Bacillus subtilis, and Escherichia coli that cause wound infection, reduce the inflammatory reaction of the wound, and can significantly improve the hemostatic speed of the trauma, and promote wound healing.

Description

Technical Field

[0001] The invention belongs to the technical field of wound treatment, and specifically relates to an external preparation for promoting wound healing and a preparation method thereof. Background Art

[0002] Trauma is the destruction of human tissue or organs caused by mechanical factors. Any external factors imposed on the human body also include structural or functional damage caused by high temperature, cold, electric current, radiation, acid, alkali, poisonous gas, poisonous insects, mosquito bites, etc. Trauma is extremely common, including: cuts, stabs, contusions, sprains. Due to the rapid development of industry, agriculture, transportation and sports, the trauma caused by various accidents is increasing. Trauma not only has a high incidence, but also varies greatly in degree. The injury can be serious and complicated, and even endanger the life of the injured. Severe trauma can cause systemic reactions, with local manifestations of pain, swelling, tenderness in the injured area; deformity and dysfunction in fractures and dislocations. Severe trauma may also cause fatal hemorrhage, shock, suffocation and impaired consciousness. Therefore, it is urgent to develop an external preparation that promotes wound hemostasis and healing, which can help people reduce the physical and economic pressure caused by pain when facing stressful events, better face difficulties, avoid negative psychological reactions, and maintain physical and mental health.

[0003] Wulingshen is the sclerotium of Xylaria nigripes (Klotzsch) Cooke, a fungus of the genus Xylaria in the family Xylariaceae. It is mainly used to treat insomnia, palpitations, hematemesis, epistaxis (hemorrhagic diseases not caused by trauma), hypertension and burns. However, there are few reports on the use of Wulingshen for trauma hemostasis and wound healing. Summary of the invention

[0004] In view of the deficiencies in the prior art, the purpose of the present invention is to provide an external preparation for promoting wound healing and a preparation method thereof. The prepared external preparation has an inhibitory effect on bacteria that cause wound infection, can increase the rate of hemostasis, and promote wound healing.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] The invention provides an external preparation for promoting wound healing, comprising the following raw materials in parts by weight: 10-20 parts of Wulingshen extract, 8-12 parts of Panax notoginseng extract, 8-12 parts of red peony root extract and 4-6 parts of borneol.

[0007] Preferably, the preparation method of the Wulingshen extract comprises: vacuum freeze-drying and crushing fresh Wulingshen, mixing it with ethanol aqueous solution and ultrasonically extracting it, filtering it, eluting the filtrate through a macroporous adsorption resin column, and concentrating and drying the eluate to obtain the Wulingshen extract.

[0008] More preferably, the volume percentage of the ethanol aqueous solution is 55%-75%.

[0009] More preferably, the solid-liquid ratio of the ultrasonic extraction is 1:4-6 g / mL, the frequency is 35-45 KHz, the temperature is 50-60° C., and the time is 20-40 min.

[0010] More preferably, the elution comprises: eluting with 35%-38% by volume ethanol aqueous solution and 49%-52% by volume ethanol aqueous solution in sequence, and collecting the eluate with 49%-52% by volume ethanol aqueous solution.

[0011] Preferably, the preparation method of the Panax notoginseng extract comprises: crushing the Panax notoginseng, mixing it with 5-10 times the volume of sesame oil, simmering it over low heat for 1-3 hours, filtering, and the filtrate is the Panax notoginseng extract.

[0012] Preferably, the preparation method of the red peony root extract comprises: crushing the red peony root, refluxing and extracting it with 8-12 times the volume of 65%-75% ethanol aqueous solution at 75-85°C for 2-4 times, each time for 2-3 hours, filtering, combining the filtrate, concentrating and drying to obtain the red peony root extract.

[0013] The present invention also provides a method for preparing the external preparation, comprising: mixing the Wulingshen extract, Panax notoginseng extract, Paeonia lactiflora extract and borneol with a solvent to obtain the external preparation; the solvent is water, polyethylene glycol 400 or glycerol.

[0014] The present invention also provides the use of the external preparation in preparing medicine for treating trauma.

[0015] Preferably, the trauma includes cuts, puncture wounds, bruises and abrasions.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The invention provides an external preparation for promoting wound healing, comprising: a Wulingshen extract, a Panax notoginseng extract, a red peony root extract and borneol. The Wulingshen extract of the invention stops bleeding and stops pain, and is the main drug; the Panax notoginseng extract removes blood stasis and stops bleeding, promotes blood circulation and stops pain, and improves the hemostatic and analgesic effect of the Wulingshen extract, and is the minister drug; the red peony root extract clears heat and cools blood, removes blood stasis and stops pain, and is the adjuvant drug; the borneol dredges meridians, and is the guiding drug; all the drugs are used together to achieve the effects of stopping blood toxicity, reducing inflammation and relieving pain, and promoting blood circulation and promoting tissue regeneration.

[0018] The present invention adjusts the method of extracting Wulingshen, and adds Panax notoginseng extract, red peony root extract and borneol on the basis thereof. The various components have synergistic effects, have inhibitory effects on Staphylococcus aureus, Bacillus subtilis and Escherichia coli that cause wound infection, reduce the inflammatory response of the wound, and can significantly improve the hemostasis speed of trauma and promote wound healing. DETAILED DESCRIPTION

[0019] The invention provides an external preparation for promoting wound healing, comprising the following raw materials in parts by weight: 10-20 parts of Wulingshen extract, 8-12 parts of Panax notoginseng extract, 8-12 parts of red peony root extract and 4-6 parts of borneol.

[0020] The vacuum freeze drying of the present invention preferably includes: first slicing the fresh black ginseng, quick freezing at -35 to -45°C for 1.5 to 2.5 hours, and vacuum freeze drying at 50 to 60 Pa and -40 to -45°C to a moisture content of 5% to 9%. The vacuum freeze drying of the present invention can retain the nutritional components of the fresh black ginseng to the greatest extent.

[0021] The particle size of the crushed material in the present invention is preferably 150-250 meshes, more preferably 200 meshes. The crushing method of the present invention can break the cell wall of the black ginseng, fully release and dissolve the effective ingredients, and improve the efficiency of subsequent ultrasonic extraction.

[0022] The volume percentage of the ethanol aqueous solution of the present invention is preferably 55%-75%, more preferably 60%; the solid-liquid ratio of the ultrasonic extraction is preferably 1:4-6g / mL, more preferably 1:5g / mL, the frequency is preferably 35-45KHz, more preferably 40KHz, the temperature is preferably 50-60°C, more preferably 55°C, and the time is preferably 20-40min, more preferably 30min. The present invention adopts the method of ethanol ultrasonic extraction, which can make the effective components of the crushed Wulingshen release more fully and improve the extraction efficiency. In the process of extracting the effective components of Wulingshen, the present invention found that compared with water extraction, the use of 55%-75% volume percentage ethanol to extract Wulingshen can significantly improve the hemostasis and healing ability of Wulingshen extract on wounds.

[0023] The macroporous adsorption resin column of the present invention is preferably a D101 macroporous adsorption resin column; the filler in the macroporous resin column is preferably 25-35 times the weight of the filtrate, and more preferably 30 times; the elution preferably includes: eluting with an ethanol aqueous solution with a volume percentage of 35%-38%, an ethanol aqueous solution with a volume percentage of 49%-52%, and collecting the eluate with a volume percentage of 49%-52% ethanol aqueous solution. More preferably, the amount of the 35%-38% ethanol aqueous solution is 5-8 times the volume of the column; the amount of the 49%-52% ethanol aqueous solution is 8-12 times the volume of the column. Further purification of the Wulingshen filtrate using a macroporous adsorption resin column can further enhance the hemostatic and healing effects of the Wulingshen extract.

[0024] The Wulingshen extract prepared by the present invention not only has good hemostatic and analgesic effects, but also has good antibacterial effects, can inhibit the growth of bacteria or other pathogens in the wound, reduce the risk of infection, alleviate inflammatory response, create favorable conditions for wound healing, and can protect the newly formed granulation tissue and epithelial cells in the wound, so that they can grow and differentiate normally, thereby accelerating the wound healing process.

[0025] The preparation method of the Panax notoginseng extract of the present invention preferably comprises: crushing Panax notoginseng, mixing with 5-10 times the volume of sesame oil, simmering for 1-3 hours on low heat, filtering, and the filtrate is the Panax notoginseng extract. More preferably, the volume is 8 times the volume, and the simmering time is 2 hours on low heat.

[0026] The invention adopts a method for extracting Panax notoginseng by slowly boiling Panax notoginseng with sesame oil, and the effective components of Panax notoginseng with the effects of removing blood stasis, stopping bleeding, promoting blood circulation and relieving pain are integrated into the sesame oil, thereby improving the hemostatic effect, and having a certain antibacterial effect, and promoting wound healing.

[0027] The preparation method of the red peony root extract of the present invention comprises: crushing the red peony root, refluxing and extracting 2-4 times with 8-12 times the volume of 65%-75% ethanol aqueous solution at 75-85°C for 2-3 hours each time, filtering, combining the filtrate, concentrating and drying to obtain the red peony root extract. More preferably, the volume is 10 times the volume, the volume percentage of the ethanol aqueous solution is 70%, the temperature is 80°C, and the refluxing extraction is performed 3 times.

[0028] The red peony root extract of the present invention can inhibit the release of inflammatory factors, relieve the redness, swelling, heat and pain of the wound, and create favorable conditions for subsequent healing; it can promote blood flow in the wound site, improve blood circulation, provide sufficient oxygen and nutrients to the tissue, accelerate cell metabolism and repair, and thus promote wound healing; it can remove free radicals generated by damage in the wound site, reduce the damage of oxidative stress to cells and tissues, protect healthy cells around the wound, and promote normal wound healing.

[0029] The borneol of the present invention can reduce the inflammatory response of the wound and relieve the pain symptoms; it has a certain inhibitory effect on various bacteria and fungi, helps to prevent wound infection, and promotes the cleaning and healing of the wound; it improves the local blood circulation of the wound, provides more nutrition and oxygen to the wound, and accelerates the repair and regeneration of tissues; it makes the area around the wound feel cool, relieves itching and discomfort, and reduces the secondary damage caused by scratching the wound; it helps to promote the absorption of other drug components through the skin, and improves the bioavailability of the drug.

[0030] The Wulingshen extract of the present invention stops bleeding and stops pain, and is the main drug. The Panax notoginseng extract removes blood stasis and stops bleeding, promotes blood circulation and stops pain, and improves the hemostatic and analgesic effect of the Wulingshen extract, and is the ministerial drug. The red peony root extract clears heat and cools blood, disperses blood stasis and stops pain, and is the adjuvant. Borneol dredges meridians and is the guiding drug. The combination of the drugs has the effects of stopping blood toxicity, reducing inflammation and relieving pain, and promoting blood circulation and promoting muscle growth. The Wulingshen extract, the Panax notoginseng extract, the red peony root extract and the borneol of the present invention have a synergistic effect, so that the external preparation has better hemostasis and wound healing effects.

[0031] The present invention also provides a method for preparing the external preparation, comprising: mixing the Wulingshen extract, Panax notoginseng extract, Paeonia lactiflora extract and borneol with a solvent to obtain the external preparation; the solvent is water, polyethylene glycol 400 or glycerol.

[0032] The mass fraction of the Wulingshen extract, Panax notoginseng extract, red peony root extract and borneol in the external preparation of the invention is 0.5%-3%.

[0033] The present invention also provides the use of the external preparation in preparing a medicine for treating trauma, wherein the trauma includes cuts, punctures, contusions and abrasions.

[0034] The method of using the topical preparation of the present invention is as follows: when applied to a wound, it is used at least three times a day, once in the morning, noon and evening. During each use, it is sprayed or applied three times continuously, once every 5 minutes, and it needs to be shaken before each use before spraying or applying.

[0035] In the present invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0036] The technical solutions in the present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0037] In the following embodiments, unless otherwise specified, all of them are conventional methods.

[0038] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0039] Example 1

[0040] Preparation of external preparations for promoting wound healing

[0041] (1) Preparation of Wulingshen Extract

[0042] Slice fresh black ginseng, quickly freeze at -40°C for 2h, freeze-dry under vacuum at 55Pa and -43°C until the moisture content is about 7.0wt%, and grind through a 200-mesh sieve to obtain black ginseng powder;

[0043] 10 g of the above-mentioned Wulingshen powder was mixed with 50 mL of 60% by volume ethanol aqueous solution, and ultrasonically extracted for 30 min at a frequency of 40 KHz and a temperature of 55° C., and filtered to obtain a filtrate;

[0044] The filtrate was passed through a D101 macroporous adsorption resin column (the filler was 30 times the weight of the filtrate), and eluted with 6 column volumes of 36% ethanol aqueous solution, and then eluted with 10 column volumes of 50% ethanol aqueous solution, and the eluate was collected;

[0045] The eluate was concentrated to 1 / 4 of its volume at 55°C, then quickly frozen at -50°C for 2h, and vacuum freeze-dried at 55Pa and -40°C to a water content of about 7.0wt%, to obtain the Wulingshen extract.

[0046] (2) Preparation of Panax notoginseng extract

[0047] The dried Panax notoginseng was crushed and passed through a 200-mesh sieve to obtain Panax notoginseng powder. 10 g of Panax notoginseng powder was mixed with 80 mL of sesame oil, and the mixture was simmered over low heat for 2 h. The mixture was filtered, and the filtrate obtained was the Panax notoginseng extract.

[0048] (3) Preparation of red peony root extract

[0049] The dried Panax notoginseng was crushed and passed through a 200-mesh sieve to obtain red peony root powder; 10 g of red peony root powder was refluxed and extracted with 100 mL of 70% by volume ethanol aqueous solution at 80° C. for 3 times, each time for 2.5 h, the filtrates were combined and concentrated to 1 / 4 of the volume of the filtrate, then quickly frozen at -50° C. for 2 h, and vacuum freeze-dried at 55 Pa and -40° C. to a moisture content of about 7.1%, to obtain the red peony root extract.

[0050] (4) Preparation of external preparations

[0051] 1.5 g of the Wulingshen extract from step (1), 1 g of the Panax notoginseng extract from step (2), 1 g of the red peony root extract from step (3) and 0.5 g of borneol were mixed with 396 g of polyethylene glycol 400 to obtain the external preparation for promoting wound healing.

[0052] Example 2

[0053] Preparation of external preparations for promoting wound healing

[0054] (1) Preparation of Wulingshen Extract

[0055] After fresh black ginseng is sliced, it is quickly frozen at -35°C for 2.5h, and vacuum freeze-dried at 50Pa and -45°C to a moisture content of about 5.2wt%, and then crushed through a 150-mesh sieve to obtain black ginseng powder;

[0056] 10 g of the above-mentioned Wulingshen powder was mixed with 40 mL of 75% by volume ethanol aqueous solution, and ultrasonically extracted for 20 min at a frequency of 35 KHz and a temperature of 60° C., and filtered to obtain a filtrate;

[0057] The filtrate was passed through a D101 macroporous adsorption resin column (the filler was 25 times the weight of the filtrate), and eluted with 5 times the column volume of a 38% ethanol aqueous solution, and then eluted with 8 times the column volume of a 52% ethanol aqueous solution, and the eluate was collected;

[0058] The eluate was concentrated to 1 / 3 of the volume of the eluate at 60°C, then quick-frozen at -45°C for 2.5h, and vacuum freeze-dried at 50Pa and -40°C to a moisture content of 5% to obtain the Wulingshen extract.

[0059] (2) Preparation of Panax notoginseng extract

[0060] The dried Panax notoginseng was crushed and passed through a 150-mesh sieve to obtain Panax notoginseng powder. 10 g of Panax notoginseng powder was mixed with 50 mL of sesame oil, and the mixture was simmered over low heat for 1 h. The mixture was filtered, and the filtrate obtained was the Panax notoginseng extract.

[0061] (3) Preparation of red peony root extract

[0062] The dried Panax notoginseng was crushed and passed through a 150-mesh sieve to obtain red peony root powder; 10 g of red peony root powder was refluxed with 80 mL of 75% by volume ethanol aqueous solution at 85° C. for 4 times, each time for 2 h, the filtrates were combined and concentrated to 1 / 3 of the volume of the filtrate, then quick-frozen at -45° C. for 2.5 h, and vacuum freeze-dried at 50 Pa and -40° C. to a moisture content of 5.4 wt% to obtain the red peony root extract.

[0063] (4) Preparation of external preparations

[0064] Mix 1 g of the Wulingshen extract from step (1), 0.8 g of the Panax notoginseng extract from step (2), 0.8 g of the red peony root extract from step (3) and 0.4 g of borneol with 97 g of polyethylene glycol 400 to obtain the external preparation for promoting wound healing.

[0065] Example 3

[0066] Preparation of external preparations for promoting wound healing

[0067] (1) Preparation of Wulingshen Extract

[0068] After fresh black ginseng is sliced, it is quickly frozen at -45°C for 1.5h, and vacuum freeze-dried at 60Pa and -40°C to a moisture content of 9%, and then crushed through a 250-mesh sieve to obtain black ginseng powder;

[0069] 10 g of the above-mentioned Wulingshen powder was mixed with 60 mL of 55% by volume ethanol aqueous solution, and ultrasonically extracted for 40 min at a frequency of 45 KHz and a temperature of 50° C., and filtered to obtain a filtrate;

[0070] The filtrate was passed through a D101 macroporous adsorption resin column (the filler was 35 times the weight of the filtrate), and eluted with 8 times the column volume of a 35% ethanol aqueous solution, and then eluted with 12 times the column volume of a 49% ethanol aqueous solution, and the eluate was collected;

[0071] The eluate was concentrated to 1 / 5 of its volume at 50°C, then quick-frozen at -50°C for 1.5h, and vacuum freeze-dried at 60Pa and -35°C to a water content of 9.3wt%, to obtain the Wulingshen extract.

[0072] (2) Preparation of Panax notoginseng extract

[0073] The dried Panax notoginseng was crushed and passed through a 250-mesh sieve to obtain Panax notoginseng powder. 10 g of Panax notoginseng powder was mixed with 100 mL of sesame oil, and the mixture was simmered over low heat for 3 h. The mixture was filtered and the filtrate obtained was the Panax notoginseng extract.

[0074] (3) Preparation of red peony root extract

[0075] The dried Panax notoginseng was crushed and passed through a 250-mesh sieve to obtain red peony root powder; 10 g of red peony root powder was refluxed with 120 mL of 65% by volume ethanol aqueous solution at 75° C. for 4 times, each time for 3 hours, the filtrates were combined and concentrated to 1 / 5 of the volume of the filtrate, then quick-frozen at -50° C. for 1.5 hours, and vacuum freeze-dried at 60 Pa and -35° C. to a moisture content of 9.4 wt% to obtain the red peony root extract.

[0076] (4) Preparation of external preparations

[0077] 2 g of the Wulingshen extract from step (1), 1.2 g of the Panax notoginseng extract from step (2), 1.2 g of the red peony root extract from step (3) and 0.6 g of borneol were mixed with 995 g of glycerol to obtain the external preparation for promoting wound healing.

[0078] Comparative Example 1

[0079] The specific implementation method is the same as that of Example 1, except that (4) the preparation of the topical preparation is as follows: 1 g of the notoginseng extract of step (2), 1 g of the red peony root extract of step (3) and 0.5 g of borneol are mixed with 247.5 g of polyethylene glycol 400 to obtain the topical preparation that promotes wound healing.

[0080] Comparative Example 2

[0081] The specific implementation method is the same as that of Example 1, except that (4) the preparation of the external preparation is to mix 1.5 g of the Urinary Glycyrrhiza uralensis extract of step (1), 1 g of the Paeonia lactiflora extract of step (3) and 0.5 g of borneol with 297 g of polyethylene glycol 400 to obtain the external preparation for promoting wound healing.

[0082] Comparative Example 3

[0083] The specific implementation method is the same as that of Example 1, except that (4) the preparation of the external preparation is to mix 1.5 g of the Urinary Glycyrrhiza uralensis extract of step (1), 1 g of the Panax notoginseng extract of step (2) and 0.5 g of borneol with 297 g of polyethylene glycol 400 to obtain the external preparation for promoting wound healing.

[0084] Comparative Example 4

[0085] The specific implementation method is the same as that of Example 1, except that (4) the preparation of the external preparation is to mix 1.5 g of the Urinary Glycyrrhiza uralensis extract of step (1), 1 g of the Panax notoginseng extract of step (2) and 1 g of the Paeonia lactiflora extract of step (3) with 346.5 g of polyethylene glycol 400 to obtain the external preparation that promotes wound healing.

[0086] Comparative Example 5

[0087] The specific implementation method is the same as that of Example 1, except that (1) the preparation of the black ginseng extract is as follows: fresh black ginseng is sliced, quickly frozen at -40°C for 2 hours, vacuum freeze-dried at 55Pa and -43°C to a moisture content of 7%, and crushed through a 200-mesh sieve to obtain black ginseng powder; 10g of the above-mentioned black ginseng powder is mixed with 50mL of pure water at room temperature and soaked for 4 hours, filtered, and the filter residue is again soaked with 50mL of pure water at room temperature for 4 hours, filtered, and the filtrate is combined, concentrated at 55°C to 1 / 4 of the volume of the eluate, quickly frozen at -50°C for 2 hours, and vacuum freeze-dried at 55Pa and -40°C to a moisture content of 7% to obtain the black ginseng extract.

[0088] Comparative Example 6

[0089] The external preparation was prepared by mixing 1 g of the Urumqi ginseng extract prepared in step (1) of Example 1 with 99 g of polyethylene glycol 400.

[0090] Comparative Example 7

[0091] The external preparation is obtained by mixing 1 g of the Urumqi ginseng extract of step (1) of Comparative Example 5 and 99 g of polyethylene glycol 400.

[0092] Experimental Example 1

[0093] Antibacterial effect of topical preparations for promoting wound healing

[0094] During the process of wound formation, bacteria from the outside world will enter the skin and cause infection, which in turn leads to bacterial infection in the wound. Common bacteria that cause wound infection usually include Staphylococcus aureus, Bacillus subtilis, Escherichia coli, etc.

[0095] Staphylococcus aureus (ATCC6538), Bacillus subtilis (ATCC6633) and Escherichia coli (ATCC25922) were used as bacteria that infect wounds, and an inhibition zone experiment was performed to verify the antibacterial effect of the external preparation for promoting wound healing of the present invention.

[0096] The antibacterial effects of the external preparations in Examples 1-3 and Comparative Examples 1-7 on Staphylococcus aureus, Bacillus subtilis and Escherichia coli were tested respectively. The specific results are shown in Table 1.

[0097] The inhibition zone test method is:

[0098] Staphylococcus aureus, Bacillus subtilis, and Escherichia coli were activated to obtain a concentration of about 5.2×10 7 CFU / mL of bacterial suspension, streak inoculated on agar medium plate, incubate in 37℃ incubator for 24h, streak inoculated on agar slant medium test tube with typical colonies, incubate in 37℃ incubator for 24h. Wash the obtained colonies repeatedly with sterile water and shake until the colonies are free of agglomeration and evenly dispersed, and dilute to about 3.4×10 7 CFU / mL bacterial suspension. 200 μL of bacterial suspension was applied to beef extract peptone medium. 5 μL of external preparations in Examples 1-3 and Comparative Examples 1-7 were dropped onto 6 mm sterile filter paper, and then the filter paper was placed on the coated medium, 3 pieces per dish, Staphylococcus aureus was inverted and cultured at 37°C for 48 hours, Bacillus subtilis and Escherichia coli were inverted and cultured for 24 hours, and the diameter of each inhibition zone in the culture dish was measured by the cross method.

[0099] Table 1 Diameters of inhibition zones of different topical preparations

[0100] Group Staphylococcus aureus (mm) Bacillus subtilis (mm) Escherichia coli (mm) Example 1 12.8 9.4 11.4 Example 2 14.5 10.6 13.3 Example 3 11.7 9.1 10.2 Comparative Example 1 9.5 8.1 8.6 Comparative Example 2 9.8 8.5 8.9 Comparative Example 3 9.8 8.7 9.2 Comparative Example 4 10.2 8.9 9.9 Comparative Example 5 9.3 7.5 9.0 Comparative Example 6 8.3 7.2 7.7 Comparative Example 7 7.2 6.3 6.4

[0101] As shown in Table 1, the external preparations of Examples 1-3 and Comparative Examples 1-7 have different degrees of inhibitory effects on Staphylococcus aureus, Bacillus subtilis, and Escherichia coli. In Examples 1-3, due to the high concentration of the active ingredient in Example 2, the antibacterial effect on Staphylococcus aureus, Bacillus subtilis, and Escherichia coli is the most obvious. Relative to Comparative Examples 1-4, Example 1 has a higher antibacterial effect, indicating that the components of the Wulingshen extract, Panax notoginseng extract, red peony root extract, and Borneol of the present invention have a synergistic effect. Relative to Comparative Example 5, Example 1 has a stronger antibacterial effect, and relative to Comparative Example 7, Comparative Example 6 has a stronger antibacterial effect, indicating that under the same active ingredient concentration, the Wulingshen extract obtained by the preparation method of the present invention can better inhibit bacterial growth and improve the wound healing speed. The above results show that the external preparation of the present invention has good antibacterial properties for Staphylococcus aureus, Bacillus subtilis, and Escherichia coli. It can be used to inhibit bacterial growth after trauma, prevent bacteria from causing inflammatory reactions to wounds, and promote wound healing.

[0102] Experimental Example 2

[0103] Hemostatic effect of topical preparations to promote wound healing

[0104] SPF grade SD male rats weighing 290-328g were selected and randomly divided into 11 groups, with 10 rats in each group. There was no significant difference between the rats in each group, so the groups were comparable.

[0105] After anesthesia, the rats in each group were fixed in the dorsal position on the experimental table. After the hair on the neck was cut off, the rats were disinfected with a 75% ethanol aqueous solution by volume, the right carotid artery was separated, the distal end was ligated, the proximal segment of the common carotid artery was clamped with an artery clamp, the blood vessel was punctured with an injection needle, the artery clamp was opened, and blood was sprayed freely for 30 seconds. The proximal blood vessels of the wound were pressed to stop bleeding. After the gauze of the external preparations of Examples 1-3 and Comparative Examples 1-7 was completely soaked, the wound was pressed, and the hemostasis was stopped by pressing. The pressure was pressed for 30 seconds within the first minute, and the hemostasis was observed. The hemostasis was observed every 1 minute for the next 3 minutes. When the bleeding still could not be stopped, the gauze soaked with the external preparation was used to repeat the above action to stop bleeding, and then the rats were observed for 2 hours to observe whether they died. The specific results are shown in Table 2.

[0106] Table 2 Hemostatic effects of different topical preparations

[0107] Group Gauze usage times (times) Coagulation time (s) Gauze weight difference before and after hemostasis (g) blank 0 300 2.8 Example 1 1 30 0.4 Example 2 1 40 0.6 Example 3 1 50 0.8 Comparative Example 1 1 100 1.2 Comparative Example 2 1 90 1.0 Comparative Example 3 1 80 0.9 Comparative Example 4 1 60 0.8 Comparative Example 5 1 160 1.4 Comparative Example 6 2 200 1.8 Comparative Example 7 2 230 2.0

[0108] As can be seen from the data in Table 2, relative to the blank control, Examples 1-3 and Comparative Examples 1-7 can shorten the clotting time and reduce the amount of bleeding. Relative to Comparative Examples 1-4, Example 1 can shorten the clotting time and reduce the amount of bleeding, indicating that the components of the Wulingshen extract, Panax notoginseng extract, red peony root extract and Borneol of the present invention have a synergistic effect in promoting blood coagulation. Relative to Comparative Example 5, Example 1 can shorten the clotting time, and relative to Comparative Example 7, Comparative Example 6 can shorten the clotting time, indicating that under the same concentration of active ingredients, the Wulingshen extract obtained by the preparation method of the present invention can better promote blood coagulation, reduce bleeding, and improve wound healing speed. The above results show that the external preparation of the present invention can significantly shorten the clotting time, reduce the amount of bleeding from the wound, and thus accelerate the healing speed of the wound.

[0109] Experimental Example 3

[0110] Wound healing effects of topical preparations that promote wound healing

[0111] SPF SD rats weighing 290-328 g were selected and randomly divided into 11 groups, with 10 rats in each group. There was no significant difference between the rats in each group, so the groups were comparable.

[0112] Use hair clippers to shave a large area of ​​the back of the test animal, and then clean it with clean water without special treatment. The next day, the test animal was anesthetized and placed in a prone position. A 10×18mm rectangular frame of the rat was drawn in the center of the back where the hair had been shaved. Use sterile surgical scissors to cut off the skin of the rectangular frame. The wound was deep to the fascia and was wiped with a sterile cotton ball to stop bleeding.

[0113] After wiping with sterile cotton balls to stop bleeding, the external preparations of Examples 1-3 and Comparative Examples 1-7 were sprayed on the defective parts after excision, and the control group was sprayed with an equal amount of polyethylene glycol 400 on the experimental wounds of the defective parts prepared in the same manner.

[0114] The wound healing effect is determined by the size and time of wound reduction. The wound size is measured by placing the experimental animal in a prone position after skin excision under general anesthesia, attaching a glass slide to the surface of the wound, and drawing the outline of the wound with a permanent marker. The glass slide with this outline is placed on a film viewing device, placed on a recording paper and marked with the glass slide record. The size of the left and right sides and the upper and lower bottom of the glass slide can be recorded to measure the degree of wound healing. The day when the wound is formed and the test agent is sprayed is defined as the first day of the experiment, and the time of starting use is recorded. After 24 hours of use, the healing effect is calculated based on the wound size and time. In order to avoid contamination with other animals, the animals participating in the experiment were kept separately. During the experiment, the cages and feeders were pre-cleaned and steam-sterilized and dried. There was no difference except for the different test solutions used.

[0115] After 4 hours, each group sprayed the wound with the same proportion of solution again, and after 8 hours, each group sprayed the wound with the same proportion of solution again. The above operation is three spraying administrations on the first day. The three spraying administrations on the first day were repeated on the second and third days. The size of the wound in each group was recorded at 12, 24, 48, 72, and 96 hours. The specific results are shown in Table 3.

[0116] Table 3 Wound healing effects of different topical preparations (mm×mm)

[0117] Group 12 hours 24 hours 48 hours 72 hours 96 hours Example 1 6×11.5 4.5×8 2×3 Less than 1×1 Less than 1×1 Example 2 6×12 5×9 3×6 Less than 1×1 Less than 1×1 Example 3 8×14 7×10 5×9 2×3 Less than 1×1 Comparative Example 1 8×15 7×13 6×11 4×8 Less than 1×1 Comparative Example 2 7×15 6×11 5×10 3.5×7 Less than 1×1 Comparative Example 3 7.5×15 6.5×12 5×11 4×7.5 Less than 1×1 Comparative Example 4 6×12.5 5×10 4×8 3×5 Less than 1×1 Comparative Example 5 7×12 6.5×11.5 4.5×9 2.5×4 Less than 1×1 Comparative Example 6 8×14 7.5×10 5×9.5 3×4 Less than 1×1 Comparative Example 7 9×15 8.5×12 6×10 3×5 Less than 1×1 Control group 10×18 10×17.5 9.5×17 9×16 9×16

[0118] As can be seen from the data in Table 3, with the use of the topical preparation, the wound healing is more significant. Relative to Comparative Examples 1-4, the wound healing effect of Example 1 is the best, indicating that the components of the Wulingshen extract, Panax notoginseng extract, red peony root extract and borneol of the present invention have a synergistic effect in promoting wound healing. Relative to Comparative Example 5, the wound healing effect of Example 1 is better, and relative to Comparative Example 7, the wound healing effect of Comparative Example 6 is better, indicating that at the same concentration of active ingredients, the Wulingshen extract obtained by the preparation method of the present invention can improve the wound healing speed. The above results show that the topical preparation of the present invention has a better wound healing effect.

[0119] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. An external preparation for promoting wound healing, characterized in that The invention is composed of the following raw materials in parts by weight: 10-20 parts of Wulingshen extract, 8-12 parts of Panax notoginseng extract, 8-12 parts of red peony root extract and 4-6 parts of borneol; The preparation method of the Wulingshen extract comprises: vacuum freeze-drying and crushing fresh Wulingshen, mixing with 55%-75% ethanol aqueous solution by volume for ultrasonic extraction, filtering, passing the filtrate through a macroporous adsorption resin column for elution, concentrating and drying the eluate to obtain the Wulingshen extract; The preparation method of the Panax notoginseng extract comprises: crushing Panax notoginseng, mixing with 5-10 times the volume of sesame oil, simmering over low heat for 1-3 hours, filtering, and obtaining the filtrate as the Panax notoginseng extract; The preparation method of the red peony root extract comprises: crushing the red peony root, refluxing and extracting it with 8-12 times the volume of 65%-75% ethanol aqueous solution at 75-85° C. for 2-4 times, each time for 2-3 hours, filtering, combining the filtrate, concentrating and drying to obtain the red peony root extract.

2. The external preparation according to claim 1, characterized in that The ultrasonic extraction has a solid-liquid ratio of 1:4-6 g / mL, a frequency of 35-45 KHz, a temperature of 50-60° C., and a time of 20-40 min.

3. The external preparation according to claim 1, characterized in that The elution comprises: eluting with an ethanol aqueous solution with a volume percentage of 35%-38% and an ethanol aqueous solution with a volume percentage of 49%-52% in sequence, and collecting the eluate with a volume percentage of ethanol aqueous solution of 49%-52%.

4. The method for preparing the external preparation according to any one of claims 1 to 3, characterized in that: include: The external preparation is obtained by mixing the Wulingshen extract, Panax notoginseng extract, Paeonia lactiflora extract and borneol with a solvent; the solvent is water, polyethylene glycol 400 or glycerin.

5. Use of the external preparation according to any one of claims 1 to 3 in the preparation of a drug for treating trauma.

6. The use according to claim 5, characterized in that: Such injuries include cuts, puncture wounds, bruises and abrasions.