Repair liquid for accelerating wound healing and preventing skin scars after trauma and preparation method thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicine, and discloses a repairing liquid for accelerating wound healing and preventing skin scars after trauma and a preparation method thereof, the repairing liquid comprises the following components in parts by weight: 25-30 parts of cape jasmine, 20-30 parts of radix ophiopogonis, 15-25 parts of poria cocos, 15-25 parts of sanguisorba officinalis, 20-30 parts of cowherb seed, 15-25 parts of rhizoma bletillae, 25-35 parts of polygonum cuspidatum, 25-30 parts of peach kernel, 20-30 parts of salvia miltiorrhiza, 20-30 parts of pseudo-ginseng, 10-20 parts of astragalus membranaceus, 10-20 parts of rhizoma acori graminei and 10-20 parts of carbonized hair; wherein the garden burnet root and the cowherb seed are subjected to combined fermentation treatment through lactobacillus delbrueckii subsp. Lactis CICC 6077 and lactobacillus acidophilus CICC 6085. The repair liquid acts on keloid fibroblasts, inhibits cell proliferation and accelerates cell apoptosis, so that collagen secretion is reduced, and hyperplastic scars are reduced and even removed. Meanwhile, the traditional Chinese medicine repairing liquid is adopted for noninvasive combined prevention and treatment of scars, and the medicine has the characteristics of early intervention time, short treatment cycle, no wound, no injection, no side effect, no medical risk, infinite close clinical endpoint to normal skin and the like.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicines, in particular to a repairing liquid for accelerating wound healing and preventing skin scars after trauma and a preparation method thereof. Background Art

[0002] Skin scars refer to the abnormal healing, appearance and pathological changes of deep skin wounds caused by trauma, burns or surgery, which are manifested as local skin changes, itching, pain, functional limitations, etc. The incidence rate of scars is significantly higher in the population. Surgery, burns, and other skin injuries deep to the dermis may lead to hypertrophic scars, which are often related to severe skin trauma, uneven surgical incisions, and wound contamination. Skin hypertrophic scars destroy the patient's body beauty and function to varying degrees, disrupt the patient's daily activities, and affect their mental health. Clinically, skin scars are often treated with corticosteroid injections, laser treatment, or surgical treatment, but the effects are uncertain. Even if patients can tolerate the pain of using several methods at the same time, the results are still unsatisfactory. Therefore, it is urgent to find new therapies for the prevention and treatment of skin scars. Based on the advantages of traditional Chinese medicine, such as no trauma, no injection, no side effects, and no medical risks, this study improves the efficacy of traditional Chinese medicine through Chinese herbal fermentation technology, and after matching, a repair fluid for accelerating wound healing and preventing skin scars after trauma was developed. Summary of the invention

[0003] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a repair liquid for accelerating wound healing and preventing skin scars after trauma and a preparation method thereof, so as to solve the problems raised in the above technical background.

[0004] To achieve the above object, the present invention is implemented through the following technical solutions:

[0005] In a first aspect, the present invention provides a repair liquid for accelerating wound healing and preventing skin scars after trauma, comprising, by weight, 25-30 parts of Gardenia, 20-30 parts of Ophiopogon japonicus, 15-25 parts of Poria cocos, 15-25 parts of Sanguisorba officinalis, 20-30 parts of Vaccaria segetalis, 15-25 parts of Bletilla striata, 25-35 parts of Polygonum cuspidatum, 25-30 parts of Persicae persica, 20-30 parts of Salvia miltiorrhiza, 20-30 parts of Panax notoginseng, 10-20 parts of Astragalus membranaceus, 10-20 parts of Acorus calamus, and 10-20 parts of Sanguisorba officinalis; wherein the Sanguisorba officinalis and Vaccaria segetalis are fermented with probiotics.

[0006] Preferably, a repair liquid for accelerating wound healing and preventing skin scars after trauma comprises, by weight, 28 parts of Gardenia, 25 parts of Ophiopogon japonicus, 20 parts of Poria cocos, 20 parts of Sanguisorba officinalis, 25 parts of Vaccaria segetalis, 20 parts of Bletilla striata, 30 parts of Polygonum cuspidatum, 28 parts of Persica kernel, 25 parts of Salvia miltiorrhiza, 25 parts of Panax notoginseng, 15 parts of Astragalus, 15 parts of Acorus calamus, and 15 parts of Sanyu charcoal.

[0007] Preferably, the probiotic comprises Lactobacillus delbrueckii subsp. lactis CICC 6077 and Lactobacillus acidophilus CICC6085.

[0008] Preferably, the concentration of the bacterial solution of Lactobacillus delbrueckii subspecies CICC 6077 is 6.0×10 8 ~1.0×10 9 cfu / mL, the concentration of Lactobacillus acidophilus CICC 6085 bacterial solution was 8.0×10 8 ~2.5×10 9 cfu / mL.

[0009] Preferably, the inoculation amount of Lactobacillus delbrueckii subsp. CICC 6077 bacterial solution is 3-6% of the total mass of Sanguisorba officinalis and Vaccaria segetalis, and the inoculation amount of Lactobacillus acidophilus CICC 6085 bacterial solution is 3-6% of the total mass of Sanguisorba officinalis and Vaccaria segetalis. Wherein 3-6% refers to the volume / mass ratio, for example, the inoculation amount of Lactobacillus delbrueckii subsp. CICC 6077 bacterial solution 3% means that 100g of the total mass of Sanguisorba officinalis and Vaccaria segetalis is inoculated with 3ml of Lactobacillus delbrueckii subsp. CICC 6077 bacterial solution.

[0010] In a second aspect, the present invention provides a method for preparing the above-mentioned repair liquid for accelerating wound healing and preventing skin scars after trauma, characterized in that it comprises the following steps:

[0011] S1. Material pretreatment: according to the formula, Gardenia, Ophiopogon japonicus, Poria cocos, Sanguisorba officinalis, Vaccaria segetalis, Bletilla striata, Polygonum cuspidatum, Persica rapa, Salvia miltiorrhiza, Panax notoginseng, Astragalus membranaceus, Acorus calamus, and Sanguisorba officinalis were weighed and crushed into 100-mesh powder respectively;

[0012] S2, take all the Chinese herbal medicine powders except Sanguisorba officinalis, Vaccaria segetalis and Ophiopogon japonicus, mix them, add 8 times the amount of water and boil for 1.5 hours, filter and take the filtrate; continue to add 8 times the amount of water to the residue and boil for 1 hour, filter and take the filtrate; continue to add 8 times the amount of water to the residue, boil for 1 hour, filter and take the filtrate, discard the residue, combine the filtrates for 3 times and concentrate to a relative density of 1.15 (60°C); after concentration, bring the filtrate to room temperature, add ethanol to adjust the ethanol concentration to 70%, seal and keep the temperature at 10°C for 36 hours for alcohol precipitation, filter and take the supernatant, concentrate the supernatant until there is no alcohol taste, and obtain liquid A for standby use;

[0013] S3, prepared at a concentration of 6.0×10 8 ~1.0×10 9 cfu / mL of Lactobacillus delbrueckii subsp. lactis CICC 6077 and a concentration of 8.0×10 8 ~2.5×10 9 cfu / mL of Lactobacillus acidophilus CICC 6085 bacterial solution;

[0014] The preparation of Lactobacillus delbrueckii subspecies CICC 6077 bacterial solution is as follows: Lactobacillus delbrueckii subspecies CICC 6077 is activated according to the conventional method, and the activated Lactobacillus delbrueckii subspecies CICC 6077 is fermented in a shake flask containing 500-1000 mL of MRS liquid culture medium for 12-18 hours to obtain a concentration of 6.0×10 8 ~1.0×10 9 cfu / mL of Lactobacillus delbrueckii subsp. lactis CICC6077 bacterial liquid.

[0015] Preparation of Lactobacillus acidophilus CICC 6085 bacterial solution: Lactobacillus acidophilus CICC 6085 was activated as usual, and the activated Lactobacillus acidophilus CICC 6085 was inoculated into a shake flask containing 500-1000 mL of MRS liquid culture medium and fermented for 12-18 hours to obtain a concentration of 8.0×10 8 ~2.5×10 9 cfu / mL of Lactobacillus acidophilus CICC 6085 bacterial solution;

[0016] MRS liquid medium: casein peptone 10.0g, beef extract powder 10.0g, yeast extract powder 5.0g, glucose 20.0g, sodium acetate 5.0g, triammonium citrate 2.0g, K 2 HPO 4 2.0 g, MgSO 4 .7H 2 O 0.2g, MnSO 4 .H 2 O 0.05g, Tween 80 1.0g, distilled water 1.0L, pH 6.2-6.5, sterilize the above ingredients at 121℃ for 15min.

[0017] S4, mix the powder of Radix Sanguisorbae and Semen Vaccariae, add the bacterial solution of Lactobacillus delbrueckii subsp. CICC 6077 and the bacterial solution of Lactobacillus acidophilus CICC 6085 at an inoculum amount of 3-6% respectively, mix evenly, and ferment anaerobicly at 37° C. for 12-18 hours; after the fermentation is completed, add 3% hydrogen peroxide to inactivate the bacterial solution of Lactobacillus delbrueckii subsp. CICC 6077, Lactobacillus delbrueckii subsp. CICC 6077 and Lactobacillus acidophilus CICC 6085 to obtain a fermented product;

[0018] S5. Take the fermented product obtained in S4 and mix it with winter wheat, add 8 times the amount of 70% ethanol, reflux extraction for 45 minutes, 3 times, discard the residue, combine the filtrate and concentrate it until there is no alcohol taste, and obtain liquid B for use;

[0019] S6. Mix the liquid obtained in step S2 and liquid B obtained in step S5 to obtain a repair solution.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] 1. The repair solution of the present invention acts on keloid fibroblasts, inhibits cell proliferation, accelerates cell apoptosis, thereby reducing collagen secretion, and then reducing or even removing hypertrophic scars.

[0022] 2. The repairing liquid of the present invention has a significant effect in inhibiting hypertrophic scars, and has the functions of flattening scars, repairing scars, and accelerating the metabolism of scar connective tissue.

[0023] 3. In the present invention, the Sanguisorba officinalis and Vaccaria segetalis contained in the repair solution are fermented with specific Lactobacillus delbrueckii subspecies CICC 6077 and Lactobacillus acidophilus CICC 6085, and act synergistically with other traditional Chinese medicine ingredients. The obtained repair solution has significantly enhanced effects on inhibiting the proliferation of human keloid fibroblasts and inhibiting hypertrophic scars, which is much higher than the repair solution obtained by fermenting the Sanguisorba officinalis and Vaccaria segetalis contained in the repair solution with a single bacterial agent, or one of the two strains using other Lactobacillus delbrueckii subspecies and other Lactobacillus acidophilus together with other traditional Chinese medicine ingredients.

[0024] 4. After being synergistically fermented by Lactobacillus delbrueckii subsp. lactis CICC 6077 and Lactobacillus acidophilus CICC 6085, Sanguisorba officinalis and Vaccaria segetalis work synergistically with winter wheat, and have the effect of significantly promoting wound healing.

[0025] 5. The use of Chinese medicine repair fluid for non-invasive combined prevention and treatment of scars has the characteristics of early intervention time, short treatment cycle, no trauma, no injection, no side effects, no medical risks, and a clinical endpoint that is infinitely close to normal skin.

[0026] The repairing liquid of the present invention has the effects of reducing inflammation, reducing redness, relieving pain, accelerating wound healing, reducing the growth rate of fibroblasts, inhibiting hypertrophic scars, and stimulating the growth of sweat glands. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a bar graph showing the effects of different repair solutions on the proliferation rate of keloid fibroblasts;

[0028] Figure 2 This is a bar graph showing the effects of different repair solutions on scar thickness of rabbit ear hypertrophic scars. DETAILED DESCRIPTION

[0029] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0030] In the present invention, the gardenia, ophiopogon, tuckahoe, sanguisorba officinalis, fenugreek, bletilla striata, polygonum cuspidatum, peach kernel, salvia miltiorrhiza, panax notoginseng, astragalus, calamus and sanguinea charcoal used are all purchased from the market, and the quality requirements all meet the standards of the 2015 edition of the pharmacopoeia.

[0031] Example 1

[0032] This embodiment provides a repair liquid for accelerating wound healing and preventing skin scarring after trauma, and the preparation method thereof comprises the following steps:

[0033] S1. Material pretreatment: weigh 28 parts of Gardenia, 25 parts of Radix Ophiopogonis, 20 parts of Poria, 20 parts of Radix Sanguisorbae, 25 parts of Vaccariae Semen, 20 parts of Bletilla striata, 30 parts of Polygonum cuspidatum, 28 parts of Semen Persicae, 25 parts of Salvia miltiorrhiza, 25 parts of Panax notoginseng, 15 parts of Astragalus, 15 parts of Acorus calamus, and 15 parts of Sanyu carbonized on the basis of weight, and grind them into 100-mesh powder respectively;

[0034] S2, take all the Chinese herbal medicine powders except Sanguisorba officinalis, Vaccaria segetalis and Ophiopogon japonicus, mix them, add 8 times the amount of water and boil for 1.5 hours, filter and take the filtrate; continue to add 8 times the amount of water to the residue and boil for 1 hour, filter and take the filtrate; continue to add 8 times the amount of water to the residue, boil for 1 hour, filter and take the filtrate, discard the residue, combine the filtrates for 3 times and concentrate to a relative density of 1.15 (60°C); after concentration, bring the filtrate to room temperature, add ethanol to adjust the ethanol concentration to 70%, seal and keep the temperature at 10°C for 36 hours for alcohol precipitation, filter and take the supernatant, concentrate the supernatant until there is no alcohol taste, and obtain liquid A for standby use;

[0035] S3, Lactobacillus delbrueckii subspecies CICC 6077 bacterial liquid preparation: Lactobacillus delbrueckii subspecies CICC 6077 was activated according to the conventional method, and the activated Lactobacillus delbrueckii subspecies CICC 6077 was fermented in a shake flask filled with 800 mL MRS liquid medium for 16 hours to obtain a concentration of 9.0×10 8 cfu / mL of Lactobacillus delbrueckii subsp. lactis CICC 6077 bacterial solution. Preparation of Lactobacillus acidophilus CICC 6085 bacterial solution: Lactobacillus acidophilus CICC 6085 was activated as usual, and the activated Lactobacillus acidophilus CICC 6085 was inoculated into a shake flask containing 800 mL of MRS liquid culture medium and fermented for 16 h to obtain a concentration of 1.0×10 9cfu / mL of Lactobacillus acidophilus CICC 6085 bacterial liquid; wherein, MRS liquid medium: casein peptone 10.0g, beef extract powder 10.0g, yeast extract powder 5.0g, glucose 20.0g, sodium acetate 5.0g, ammonium citrate tribasic 2.0g, K 2 HPO 4 2.0 g, MgSO 4 .7H 2 O 0.2g, MnSO 4 .H 2 O0.05g, Tween 80 1.0g, distilled water 1.0L, pH 6.5, sterilize the above ingredients at 121℃ for 15min.

[0036] S4, take Sanguisorba officinalis and Vaccaria segetalis powder and mix them, add Lactobacillus delbrueckii subspecies CICC 6077 bacterial solution and Lactobacillus acidophilus CICC 6085 bacterial solution in an inoculation amount of 5% of the total mass of Sanguisorba officinalis and Vaccaria segetalis, respectively, mix and put into a cylinder for fermentation, anaerobic fermentation, the temperature is 37 ° C, and the fermentation time is 16 hours; after the fermentation is completed, 3% hydrogen peroxide is added to inactivate Lactobacillus delbrueckii subspecies CICC 6077, Lactobacillus delbrueckii subspecies CICC 6077 bacterial solution and Lactobacillus acidophilus CICC 6085 to obtain a fermentation product;

[0037] S5. Take the fermented product obtained in S4 and mix it with winter wheat, add 8 times the amount of 70% ethanol, reflux extraction for 45 minutes, 3 times, discard the residue, combine the filtrate and concentrate it until there is no alcohol taste, and obtain liquid B for use;

[0038] S6. Mix the liquid obtained in step S2 and liquid B obtained in step S5 to obtain a repair solution.

[0039] Comparative Example 1

[0040] This comparative example is similar to Example 1, except that only Lactobacillus delbrueckii subspecies CICC6077 is used for fermentation in step S4; the rest is the same as Example 1.

[0041] Comparative Example 2

[0042] This comparative example is similar to Example 1, except that only Lactobacillus acidophilus CICC 6085 is used for fermentation in step S4; the rest is the same as Example 1.

[0043] Comparative Example 3

[0044] This comparative example is similar to Example 1, except that in step S4, other Lactobacillus delbrueckii subspecies lactis and Lactobacillus acidophilus CICC 6085 are used for fermentation, and the other Lactobacillus delbrueckii subspecies lactis is Lactobacillus delbrueckii subspecies CICC 25164, which is purchased from China Industrial Microbiological Culture Collection Center; the rest is the same as Example 1.

[0045] Comparative Example 4

[0046] This comparative example is similar to Example 1, except that in step S4, Lactobacillus delbrueckii subspecies CICC 6077 and other Lactobacillus acidophilus are used for fermentation, and the other Lactobacillus acidophilus is Lactobacillus acidophilus CICC 6096 purchased from China Industrial Microbiological Culture Collection Center; the rest is the same as Example 1.

[0047] Comparative Example 5

[0048] This comparative example is similar to Example 1, except that the Sanguisorba officinalis and Vaccaria segetalis in step S4 are not fermented, but are directly mixed with winter wheat for alcohol extraction in step S5; the rest is the same as Example 1.

[0049] 1. The inhibitory effect of repair fluid on keloid fibroblasts

[0050] 1. CCK-8 assay to detect cell proliferation activity

[0051] 1.1 Test samples and grouping: 6 test groups and 1 negative control group were set up, and the test groups respectively used the repair solutions of Example 1 and Comparative Examples 1-5 of the present application. Human keloid fibroblasts, product number: FHHUM184, purchased from Hunan Fenghui Biotechnology Co., Ltd.;

[0052] 1.2 Experimental method: Human keloid fibroblasts were cultured and prepared at a concentration of 1×10 5 Each well of the suspension was inoculated with 100ul per well in a 96-well plate, with 5 replicates per group. 2 Under the conditions of 5% PBS, the cells were cultured overnight. After the cells adhered, the culture medium was discarded. After washing with PBS, the culture medium containing 0.5% v / v of the repair solution corresponding to Example 1 and Comparative Examples 1-5 was added (the culture medium was a DMEM culture medium containing 0.5% fatty acid-free bovine serum albumin and 10% fetal bovine serum), and a DMEM culture medium containing 0.5% fatty acid-free bovine serum albumin and 10% fetal bovine serum without adding drugs was set as a negative control group. After continuing to culture for 24 hours, the culture medium in the well was discarded. After washing with PBS, 20 μL of a mixed solution of MTT reagent (5 mg / mL, PBS configuration) and 180 μL of serum-free DMEM was added to each well. At the same time, a zeroing well (without cells, 20 μL MTT+180 μL serum-free DMEM culture medium) was set as a blank control. After incubation in a cell culture incubator for 4 hours, the culture medium was discarded, 150 μL DMSO was added, and the cells were shaken for 10 minutes to fully dissolve the crystals. The absorbance value of each well was measured at a wavelength of 490 nm by an ELISA instrument to calculate the cell proliferation rate. The cell proliferation rate is shown in Tables 1 and 2. Figure 1 As shown;

[0053] Cell proliferation rate = (OD value of the experimental group - OD value of the blank control group) / (OD value of the negative control group - OD value of the blank control group) × 100%, where:

[0054] OD value of the test group: the absorbance value of the cultured cell group of the treatment group (Example 1, Comparative Examples 1-5).

[0055] OD value of negative control group: absorbance value of the cell group cultured normally without adding drugs.

[0056] OD value of blank control group: absorbance value of zero-adjusted well without cells.

[0057] Table 1

[0058]

[0059]

[0060] From Table 1 and Figure 1 It can be seen that the repair solutions provided in Example 1 and Comparative Examples 1-5 can inhibit the proliferation of human keloid fibroblasts. Compared with Comparative Examples 1-5, the cell proliferation rate of human keloid fibroblasts cultured with the repair solution provided in Example 1 is significantly reduced, indicating that the repair solution provided in Example 1 can more significantly inhibit the proliferation of human keloid fibroblasts, thereby reducing collagen secretion, and then reducing or even removing hypertrophic scars.

[0061] 2. Effects of Repair Cream on Hypertrophic Scars

[0062] 2.1 Experimental grouping: 42 New Zealand white rabbits with healthy ears were randomly divided into 7 groups, 6 rabbits in each group, namely the model group, Example 1 group, and Comparative Examples 1-5 groups;

[0063] 2.2 Test methods:

[0064] Rabbit ear hypertrophic scar modeling: 42 New Zealand white rabbits with healthy ears were selected, each of which was housed in a single cage under standard conditions and fed adaptively for 1 week. After anesthesia and coma, the rabbit ear abdomen was routinely disinfected with iodine tincture, and three circular wounds with a diameter of 1 cm were punched along both sides of the rabbit ear abdomen with a puncher, and the distance between each wound was greater than 5 mm. The skin and subcutaneous tissue were cut along the wound with a sterile blade, and the perichondrium was carefully peeled off with forceps and ophthalmic scissors to retain the cartilage, and disinfected again, and sterile gauze was used to press and stop bleeding. The gauze was removed after 1 day, and the wound secretions were cleaned.

[0065] One week after modeling, the wound surface was epithelialized, and the rabbit ear wound surface was treated. From the 8th day after surgery, the model group was not treated, and the Example 1 group and the Comparative Example 1-5 groups were smeared with the corresponding repair solution, once a day, 100ul / cm 2After 4 weeks of treatment, the wounds on the rabbit ears of each group were observed. At the same time, the large-eared white rabbits were anesthetized and killed, and the scar tissues of the rabbit ears of each group were cut from the surface scar tissue to the cartilage layer. The thickness of the scars in each group was measured, recorded and statistically analyzed. The results are shown in Table 2 and Figure 2 shown.

[0066] Table 2

[0067] Group Rabbit ear scar thickness / mm Model Group 11.3±2.6 Example 1 1.8±1.5 Comparative Example 1 6.9±2.0 Comparative Example 2 7.1±1.9 Comparative Example 3 6.5±2.2 Comparative Example 4 6.2±1.8 Comparative Example 5 7.8±2.1

[0068] From Table 2 and Figure 2 It can be seen that after 4 weeks of treatment, the hypertrophic scars of the rabbit ears in the model group are significantly more prominent than the surrounding normal skin, with a rough surface and a tough texture, and are in the form of hard nodules; while the hypertrophic scars of the rabbit ears in the Example 1 group are significantly thinner and closer to the surrounding skin, and most of the hypertrophic scar tissues are basically as smooth as the surrounding skin. The scar thickness of all the cut rabbit ear hypertrophic scars was measured, and the results showed that compared with the model group, the scar thickness of the Example 1 group and the Comparative Examples 1-5 groups was significantly reduced, and compared with the Comparative Examples 1-5 groups, the scar thickness of the Example 1 group was significantly reduced, which shows that Example 1 can significantly inhibit the formation of hypertrophic scars in rabbit ears and has the effect of repairing scars. This is because the specific Lactobacillus lactis subspecies CICC 6077 and Lactobacillus acidophilus CICC 6085 synergistic fermentation of Sanguisorba officinalis and Vaccaria segetalis act synergistically with other Chinese medicine ingredients, and have the effect of significantly inhibiting the hypertrophic scars of rabbit ears.

[0069] As can be seen from the above, the scar prevention effect of Example 1 is significantly higher than that of Comparative Examples 1-5. This is because the Sanguisorba officinalis and Vaccaria segetalis contained in the repair solution are fermented with specific Lactobacillus delbrueckii subspecies CICC 6077 and Lactobacillus acidophilus CICC 6085, and act synergistically with other Chinese medicinal ingredients. The obtained repair solution has a significantly enhanced effect of inhibiting the proliferation of human keloid fibroblasts and inhibiting hypertrophic scars, which is much higher than the repair solution obtained by fermenting the Sanguisorba officinalis and Vaccaria segetalis contained in the repair solution with a single bacterial agent, or one of the two strains using other Lactobacillus delbrueckii subspecies and other Lactobacillus acidophilus together with other Chinese medicinal ingredients.

[0070] Example 2

[0071] This embodiment provides a repair liquid for accelerating wound healing and preventing skin scarring after trauma, and the preparation method thereof comprises the following steps:

[0072] S1. Material pretreatment: weigh 25 parts of Gardenia, 30 parts of Ophiopogon, 15 parts of Poria, 15 parts of Sanguisorba officinalis, 30 parts of Vaccaria segetalis, 15 parts of Bletilla striata, 35 parts of Polygonum cuspidatum, 25 parts of Persica, 30 parts of Salvia miltiorrhiza, 20 parts of Panax notoginseng, 20 parts of Astragalus, 20 parts of Acorus calamus, and 10 parts of Sanyu charcoal by weight, and grind them into 100-mesh powder respectively;

[0073] S2, take all the Chinese herbal medicine powders except Sanguisorba officinalis, Vaccaria segetalis and Ophiopogon japonicus, mix them, add 8 times the amount of water and boil for 1.5 hours, filter and take the filtrate; continue to add 8 times the amount of water to the residue and boil for 1 hour, filter and take the filtrate; continue to add 8 times the amount of water to the residue, boil for 1 hour, filter and take the filtrate, discard the residue, combine the filtrates for 3 times and concentrate to a relative density of 1.15 (60°C); after concentration, bring the filtrate to room temperature, add ethanol to adjust the ethanol concentration to 70%, seal and keep the temperature at 10°C for 36 hours for alcohol precipitation, filter and take the supernatant, concentrate the supernatant until there is no alcohol taste, and obtain liquid A for standby use;

[0074] S3, preparation of bacterial liquid of Lactobacillus delbrueckii subspecies CICC 6077: Lactobacillus delbrueckii subspecies CICC 6077 was activated as usual, and the activated Lactobacillus delbrueckii subspecies CICC 6077 was fermented in a shake flask containing 500 mL of MRS liquid medium for 12 h to obtain a liquid liquid with a concentration of 6.0×10 8 cfu / mL of Lactobacillus delbrueckii subsp. lactis CICC 6077 bacterial solution. Preparation of Lactobacillus acidophilus CICC 6085 bacterial solution: Lactobacillus acidophilus CICC 6085 was activated as usual, and the activated Lactobacillus acidophilus CICC 6085 was inoculated into a shake flask containing 1000 mL of MRS liquid culture medium and fermented for 18 hours to obtain a concentration of 2.5×10 9 cfu / mL of Lactobacillus acidophilus CICC 6085 bacterial liquid; wherein, MRS liquid medium: casein peptone 10.0g, beef extract powder 10.0g, yeast extract powder 5.0g, glucose 20.0g, sodium acetate 5.0g, ammonium citrate tribasic 2.0g, K 2 HPO 4 2.0 g, MgSO 4 .7H 2 O 0.2g, MnSO 4 .H 2 O0.05g, Tween 80 1.0g, distilled water 1.0L, pH 6.5, sterilize the above ingredients at 121℃ for 15min.

[0075] S4, take Sanguisorba officinalis and Vaccaria segetalis powder and mix them, add Lactobacillus delbrueckii lactis CICC 6077 solution at an inoculation amount of 6% of the total mass of Sanguisorba officinalis and Vaccaria segetalis and add Lactobacillus acidophilus CICC 6085 solution at an inoculation amount of 3% of the total mass of Sanguisorba officinalis and Vaccaria segetalis, mix them, and then put them into a cylinder for fermentation, anaerobic fermentation, the temperature is 37° C., and the fermentation time is 18 hours; after the fermentation is completed, 3% hydrogen peroxide is added to inactivate Lactobacillus delbrueckii lactis CICC 6077, Lactobacillus delbrueckii lactis CICC 6077 solution and Lactobacillus acidophilus CICC 6085 to obtain a fermentation product;

[0076] S5. Take the fermented product obtained in S4 and mix it with winter wheat, add 8 times the amount of 70% ethanol, reflux extraction for 45 minutes, 3 times, discard the residue, combine the filtrate and concentrate it until there is no alcohol taste, and obtain liquid B for use;

[0077] S6. Mix the liquid obtained in step S2 and liquid B obtained in step S5 to obtain a repair solution.

[0078] Comparative Example 6

[0079] This comparative example is similar to Example 2, except that the Sanguisorba officinalis and Vaccaria segetalis in step S4 are not fermented, but are directly mixed with winter wheat for alcohol extraction in step S5; the rest is the same as Example 2.

[0080] Comparative Example 7

[0081] This comparative example is similar to Example 2, except that Sanguisorba officinalis is removed in step S4, and only Vaccaria segetalis is used for fermentation, and Sanguisorba officinalis is subjected to alcohol extraction together with the fermentation product and winter wheat in step S5; the rest is the same as Example 2.

[0082] Comparative Example 8

[0083] This comparative example is similar to Example 2, except that in step S4, Vaccaria segetalis is removed, only Sanguisorba officinalis is used for fermentation, and Vaccaria segetalis is subjected to alcohol extraction together with the fermentation product and winter wheat in step S5; the rest is the same as Example 2.

[0084] Comparative Example 9

[0085] This comparative example is similar to Example 2, except that winter wheat is removed in step S5, and only the fermented product is used for alcohol extraction; the rest is the same as Example 2.

[0086] 3. Effect of repair fluid on wound healing

[0087] 3.1 Experimental grouping: 60 SD rats (purchased from Nanjing Junke Biotechnology Co., Ltd.), weighing 180-220 g, were raised for one week to adapt to the new environment; 10 rats in each group were randomly divided into 6 groups, including 5 drug-treated groups and 1 model group; both the model group and the drug-treated group used modeling mice, the model group was not medicated, and the 5 drug-treated groups were respectively treated with the repair solutions obtained in Example 2 and Comparative Examples 6-9;

[0088] Experimental method: After one week of adaptive feeding, the SD rats were anesthetized, the hair on the back was cut, and a 1.0 cm × 1.0 cm full-thickness skin wound model was established on the back; after the model was successfully established, the five drug-administered groups were respectively smeared with repair solution on the wounds of the modeled rats, using the corresponding repair solution obtained in Example 2 and Comparative Examples 6-9 (100 ul / cm 2 );

[0089] The wound healing conditions were observed 3, 7, 10, and 14 days after administration, and the wound healing rates of the rats in each group were calculated. The statistical results are shown in Table 3. The calculation formula of the wound healing rate is as follows:

[0090] Wound healing rate (%) = [(initial wound area - wound area on day n) / initial wound area] × 100%

[0091] Table 3 Dynamic changes of wound healing rate by repair fluid (%)

[0092]

[0093]

[0094] As shown in Table 3, there is no obvious difference between the groups after 3 days, and the healing effect of Example 2 is significantly improved compared with the model group after 7 days, 10 days and 14 days; it can be seen that the repair solution of the present invention has a significant effect on promoting the healing of rat wounds; compared with Comparative Examples 6-9, Example 2 has a significant healing effect, indicating that Sanguisorba officinalis and Vaccaria segetalis, after being synergistically fermented by Lactobacillus delbrueckii subsp. lactis CICC 6077 and Lactobacillus acidophilus CICC 6085, work synergistically with winter wheat, and have the effect of significantly promoting wound healing.

[0095] Example 3

[0096] This embodiment provides a repair liquid for accelerating wound healing and preventing skin scarring after trauma, and the preparation method thereof comprises the following steps:

[0097] S1. Material pretreatment: weigh 30 parts of Gardenia, 20 parts of Ophiopogon japonicus, 25 parts of Poria cocos, 25 parts of Sanguisorba officinalis, 20 parts of Vaccaria segetalis, 25 parts of Bletilla striata, 25 parts of Polygonum cuspidatum, 30 parts of Persicae persica, 20 parts of Salvia miltiorrhiza, 30 parts of Panax notoginseng, 10 parts of Astragalus, 10 parts of Acorus calamus, and 20 parts of Sanyu charcoal by weight, and grind them into 100-mesh powder respectively;

[0098] S2, take all the Chinese herbal medicine powders except Sanguisorba officinalis, Vaccaria segetalis and Ophiopogon japonicus, mix them, add 8 times the amount of water and boil for 1.5 hours, filter and take the filtrate; continue to add 8 times the amount of water to the residue and boil for 1 hour, filter and take the filtrate; continue to add 8 times the amount of water to the residue, boil for 1 hour, filter and take the filtrate, discard the residue, combine the filtrates for 3 times and concentrate to a relative density of 1.15 (60°C); after concentration, bring the filtrate to room temperature, add ethanol to adjust the ethanol concentration to 70%, seal and keep the temperature at 10°C for 36 hours for alcohol precipitation, filter and take the supernatant, concentrate the supernatant until there is no alcohol taste, and obtain liquid A for standby use;

[0099] S3, preparation of bacterial liquid of Lactobacillus delbrueckii subspecies CICC 6077: Lactobacillus delbrueckii subspecies CICC 6077 was activated as usual, and the activated Lactobacillus delbrueckii subspecies CICC 6077 was fermented in a shake flask containing 1000 mL of MRS liquid medium for 18 h to obtain a liquid liquid of 1.0×10 9 cfu / mL of Lactobacillus delbrueckii subsp. lactis CICC 6077 bacterial solution. Preparation of Lactobacillus acidophilus CICC 6085 bacterial solution: Lactobacillus acidophilus CICC 6085 was activated as usual, and the activated Lactobacillus acidophilus CICC 6085 was inoculated into a shake flask containing 500 mL of MRS liquid culture medium and fermented for 12 h to obtain a concentration of 8.0×10 8 cfu / mL of Lactobacillus acidophilus CICC 6085 culture medium; MRS liquid medium: casein peptone 10.0g, beef extract powder 10.0g, yeast extract powder 5.0g, glucose 20.0g, sodium acetate 5.0g, triammonium citrate 2.0g, K 2 HPO 4 2.0 g, MgSO 4 .7H 2 O 0.2g, MnSO 4 .H 2 O 0.05g, Tween 80 1.0g, distilled water 1.0L, pH 6.5, sterilize the above ingredients at 121℃ for 15min.

[0100] S4, take Sanguisorba officinalis and Vaccaria segetalis powder and mix them, add Lactobacillus delbrueckii lactis CICC 6077 solution at an inoculation amount of 3% of the total mass of Sanguisorba officinalis and Vaccaria segetalis and add Lactobacillus acidophilus CICC 6085 solution at an inoculation amount of 6% of the total mass of Sanguisorba officinalis and Vaccaria segetalis, mix them, and then put them into a cylinder for fermentation, anaerobic fermentation, temperature is 37 ° C, fermentation time is 12 hours; after fermentation is completed, add 3% hydrogen peroxide to inactivate Lactobacillus delbrueckii lactis CICC 6077, Lactobacillus delbrueckii lactis CICC 6077 solution and Lactobacillus acidophilus CICC 6085 to obtain a fermentation product;

[0101] S5. Take the fermented product obtained in S4 and mix it with winter wheat, add 8 times the amount of 70% ethanol, reflux extraction for 45 minutes, 3 times, discard the residue, combine the filtrate and concentrate it until there is no alcohol taste, and obtain liquid B for use;

[0102] S6. Mix the liquid obtained in step S2 and liquid B obtained in step S5 to obtain a repair solution.

[0103] The above-mentioned embodiments only express the specific implementation of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention.

Claims

1. A repairing liquid for accelerating wound healing and preventing skin scarring after trauma, characterized in that: The composition comprises 25-30 parts of Gardenia, 20-30 parts of Ophiopogon japonicus, 15-25 parts of Poria cocos, 15-25 parts of Sanguisorba officinalis, 20-30 parts of Vaccaria segetalis, 15-25 parts of Bletilla striata, 25-35 parts of Polygonum cuspidatum, 25-30 parts of Persicae persica, 20-30 parts of Salvia miltiorrhiza, 20-30 parts of Panax notoginseng, 10-20 parts of Astragalus membranaceus, 10-20 parts of Acorus calamus, and 10-20 parts of Sanguisorba officinalis charcoal; wherein the Sanguisorba officinalis and Vaccaria segetalis are fermented with probiotics.

2. The repairing liquid for accelerating wound healing and preventing skin scarring after trauma according to claim 1, characterized in that: The composition comprises 28 parts of Gardenia, 25 parts of Ophiopogon, 20 parts of Poria, 20 parts of Sanguisorba officinalis, 25 parts of Vaccaria segetalis, 20 parts of Bletilla striata, 30 parts of Polygonum cuspidatum, 28 parts of Persica, 25 parts of Salvia miltiorrhiza, 25 parts of Panax notoginseng, 15 parts of Astragalus, 15 parts of Acorus calamus and 15 parts of Sanguisorba officinalis.

3. The repairing liquid for accelerating wound healing and preventing skin scarring after trauma according to claim 1, characterized in that: The probiotic comprises Lactobacillus delbrueckii subsp. lactis CICC 6077 and Lactobacillus acidophilus CICC 6085.

4. The repairing liquid for accelerating wound healing and preventing skin scarring after trauma according to claim 3, characterized in that: The concentration of the Lactobacillus delbrueckii subspecies CICC 6077 bacterial solution is 6.0×10 8 ~1.0×10 9 cfu / mL, the concentration of Lactobacillus acidophilus CICC 6085 bacterial solution was 8.0×10 8 ~2.5×10 9 cfu / mL.

5. The repairing liquid for accelerating wound healing and preventing skin scarring after trauma according to claim 4, characterized in that: The inoculation amount of the bacterial solution of Lactobacillus delbrueckii subspecies CICC 6077 is 3-6% of the total weight of Sanguisorba officinalis and Vaccaria segetalis.

6. The repairing liquid for accelerating wound healing and preventing skin scarring after trauma according to claim 4, characterized in that: The inoculation amount of Lactobacillus acidophilus CICC 6085 bacterial solution is 3-6% of the total weight of Sanguisorba officinalis and Vaccaria segetalis.

7. A method for preparing a repairing liquid for accelerating wound healing and preventing skin scars after trauma according to any one of claims 1 to 6, characterized in that: The following steps are involved: S1. Material pretreatment: according to the formula, Gardenia, Ophiopogon japonicus, Poria cocos, Sanguisorba officinalis, Vaccaria segetalis, Bletilla striata, Polygonum cuspidatum, Persica rapa, Salvia miltiorrhiza, Panax notoginseng, Astragalus membranaceus, Acorus calamus, and Sanguisorba officinalis were weighed and crushed into 100-mesh powder respectively; S2. Mix all the powders of Chinese medicinal materials except Sanguisorba officinalis, Vaccaria segetalis and Olea europaea, extract with water to obtain a filtrate and concentrate to a relative density of 1.15 (60° C.); after concentration, bring the filtrate to room temperature, add ethanol to adjust the ethanol concentration to 70%, seal and keep at 10° C. for 36 hours for alcohol precipitation, filter and obtain the supernatant, concentrate the supernatant until there is no alcohol taste, and obtain liquid A for standby use; S3, prepared at a concentration of 6.0×10 8 ~1.0×10 9 cfu / mL of Lactobacillus delbrueckii subsp. lactis CICC 6077 and a concentration of 8.0×10 8 ~2.5×10 9 cfu / mL of Lactobacillus acidophilus CICC 6085 bacterial solution; S4, take Sanguisorba officinalis and Vaccaria segetalis powder and mix them, add Lactobacillus delbrueckii subspecies CICC6077 bacterial solution and Lactobacillus delbrueckii subspecies CICC 6077 bacterial solution and Lactobacillus acidophilus CICC 6085 bacterial solution at an inoculum amount of 3-6% respectively, mix them evenly, and ferment them anaerobically at 37° C. for 12-18 hours; after the fermentation is completed, add 3% hydrogen peroxide to inactivate Lactobacillus delbrueckii subspecies CICC 6077, Lactobacillus delbrueckii subspecies CICC 6077 bacterial solution and Lactobacillus acidophilus CICC 6085 to obtain a fermented product; S5. Take the fermented product obtained in S4 and mix it with winter wheat, add 8 times the amount of 70% ethanol, reflux extraction for 45 minutes, 3 times, discard the residue, combine the filtrate and concentrate it until there is no alcohol taste, and obtain liquid B for use; S6. Mix the liquid obtained in step S2 and liquid B obtained in step S5 to obtain a repair solution.

8. The method for preparing a repairing liquid for accelerating wound healing and preventing skin scarring after trauma according to claim 7, characterized in that: In step S2, the specific method of the water extraction method is: take all the powders of Chinese medicinal materials except Sanguisorba officinalis, Vaccaria segetalis and Ophiopogon japonicus, mix them, add 8 times the amount of water and boil them for 1.5 hours, filter and obtain the filtrate; continue to add 8 times the amount of water to the residue and boil them for 1 hour, filter and obtain the filtrate; continue to add 8 times the amount of water to the residue and boil them for 1 hour, filter and obtain the filtrate, discard the residue, and combine the filtrates three times to obtain the product.

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