Application of detoxifying and tissue regeneration promoting composition in preparation of medicine for treating diabetic foot ulcer

Abstract

The invention relates to application of a detoxifying and tissue regeneration promoting composition in preparation of a medicine for treating diabetic foot ulcer. The detoxifying and tissue regeneration promoting composition is prepared from the following raw materials in parts by weight: 60-100 parts of radix arnebiae seu lithospermi, 60-100 parts of radix angelicae sinensis, 20-60 parts of radix angelicae, 20-60 parts of liquorice root, 10-40 parts of frankincense and 10-50 parts of dried alum.

Description

Technical Field

[0001] The invention relates to the field of traditional Chinese medicine, and in particular to use of a detoxification and muscle-generating composition in preparing a medicine for treating diabetic foot ulcers. Background Art

[0002] Diabetic foot ulcer (DFU) is a common and serious complication in patients with diabetes. It is caused by peripheral neuropathy, vascular disease and infection. It is difficult to heal, easy to relapse and has a high risk of amputation. Many foot complications in patients with diabetes arise from sensory neuropathy and mild autonomic and motor neuropathy. Among them, sensory neuropathy combined with excessive mechanical stress is the main initiating factor for foot ulcers and infections. Inflammation and tissue damage are the result of a certain degree of repeated stress acting on a specific area of ​​loss of sensation. Pressure or shear force from the ground, shoes or other adjacent toes leads to ulcer formation. Due to the lack of normal nerve protection mechanism, ulcers are often aggravated by the presence of bone protrusions. Lesions of the autonomic nervous system cause the loss of the skin's normal sweat regulation function, skin temperature regulation function and blood supply regulation ability, resulting in reduced local tissue flexibility, the formation of thick calluses and greater brittleness and cracking. In addition, the loss of normal sweating ability blocks the rehydration of local tissues, causing further tissue damage and making deep tissues more susceptible to bacterial colonization. Motor neuropathy also plays a role in the pathogenesis of diabetic foot, and contracture of the intrinsic muscles of the foot causes the typical claw-toe deformity. Hyperextension of the metatarsophalangeal joint has also been shown to directly increase pressure under the metatarsal heads, making this area more susceptible to ulceration. Flexion of the proximal interphalangeal joint increases the risk of ulceration on the dorsal side of the protruding interphalangeal joint and the plantar side of the toe tip, while vascular disease makes it difficult for damaged tissue to heal.

[0003] At present, the dressings used for diabetic foot ulcers are selected from hydrogels (dry wounds), alginate (high exudate) or silver-containing dressings (anti-infection) according to the characteristics of the wound surface.

[0004] The detoxification and granulation composition is composed of lithospermum officinale, angelica sinensis, angelica dahurica, liquorice, frankincense and alum, and has the effects of promoting blood circulation and dispersing blood stasis, relieving swelling and relieving pain, detoxifying and removing pus, removing dead tissue and granulation. The inventor has found through research that the detoxification and granulation composition as a dressing has a good therapeutic effect on diabetic foot ulcers. Summary of the invention

[0005] The purpose of the present invention is to provide a detoxification and muscle-generating composition for use in preparing a medicine for treating diabetic foot ulcers.

[0006] The detoxification and muscle-building composition is made of the following raw materials in parts by weight:

[0007] 60-100 parts of lithospermum, 60-100 parts of angelica, 20-60 parts of angelica dahurica, 20-60 parts of liquorice, 10-40 parts of frankincense and 10-50 parts of alum.

[0008] Preferably, the detoxification and muscle-building composition is made from the following raw materials in parts by weight:

[0009] 70-90 parts of lithospermum officinale, 70-90 parts of angelica sinensis, 30-50 parts of angelica dahurica, 30-50 parts of liquorice, 20-30 parts of vinegar-processed frankincense, and 30-40 parts of alum.

[0010] Preferably, the method for preparing the active ingredient of the detoxifying and muscle-regenerating composition comprises the following steps:

[0011] Take frankincense and alum and grind them into fine powder respectively, then sieve them;

[0012] Soaking lithospermum officinale, angelica sinensis, angelica dahurica and licorice in 350-1000 parts by weight of tea oil for 5-10 days, stirring 1-3 times a day, and then boiling at 105-110° C. for 0.5-2 hours to obtain active ingredient oil;

[0013] Frankincense, Alum and Active Ingredient Oil are the active ingredients.

[0014] Preferably, the method for preparing the active ingredient of the detoxifying and muscle-regenerating composition comprises the following steps:

[0015] Take frankincense and alum and grind them into fine powder respectively, then sieve them;

[0016] Soaking lithospermum officinale, angelica sinensis, angelica dahurica and licorice in 400-700 parts by weight of tea oil for 5-10 days, stirring 1-3 times a day, and then boiling at 105-110° C. for 0.5-2 hours to obtain active ingredient oil;

[0017] Frankincense, Alum and Active Ingredient Oil are the active ingredients.

[0018] Preferably, the preparation method of the ointment of the detoxifying and muscle-promoting composition comprises the following steps:

[0019] Soak lithospermum officinale, angelica sinensis, angelica dahurica and licorice in 350-1000 parts by weight of tea oil for 5-10 days, stir 1-3 times a day, then boil at 105-110° C. for 0.5-2 hours, add 60-200 parts by weight of matrix to dissolve, filter, add frankincense and alum powder to the filtrate, mix well, and obtain;

[0020] The matrix is ​​one or a combination of beeswax, lanolin, vaseline, polyethylene glycol and white wax.

[0021] Preferably, the preparation method of the ointment of the detoxifying and muscle-promoting composition comprises the following steps:

[0022] Soak lithospermum officinale, angelica sinensis, angelica dahurica and licorice in 400-700 parts by weight of tea oil for 5-10 days, stir 1-3 times a day, then boil at 105-110° C. for 0.5-2 hours, add 80-120 parts by weight of matrix to dissolve, filter, add frankincense and alum powder to the filtrate, mix well, and obtain;

[0023] The matrix is ​​one or a combination of beeswax, lanolin, vaseline, polyethylene glycol and white wax.

[0024] The present invention also discloses a Chinese medicine preparation for treating diabetic foot ulcer, which is prepared by combining the above-mentioned detoxification and muscle-generating composition with a pharmaceutically acceptable carrier. Various commonly used preparations that can be prepared include but are not limited to: tinctures, syrups, sprays, ointments, plasters, papules, gels, etc.

[0025] Pharmacodynamic studies have shown that the detoxifying and myogenic composition of the present invention can significantly improve the healing rate of wounds of rats with diabetic foot ulcers, has an increasing trend on the average blood perfusion of rat wounds, and affects the content and expression of VEGF. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Effects of JDSJ on blood glucose in SD rats with diabetic foot ulcer model (n=10); Note: Compared with the blank control group, # indicates significant difference (P<0.05), ## indicates significant difference (P<0.01), and ### indicates significant difference (P<0.001); compared with the model control group, * indicates significant difference (P<0.05), ** indicates significant difference (P<0.01), and *** indicates significant difference (P<0.001).

[0027] Figure 2 The effect of JDSJ on the healing rate of diabetic foot ulcer model SD rats (n=10, %); Note: Compared with the blank control group, # indicates a significant difference (P<0.05), ## indicates a significant difference (P<0.01), and ### indicates a significant difference (P<0.001); Compared with the model control group, * indicates a significant difference (P<0.05), ** indicates a significant difference (P<0.01), and *** indicates a significant difference (P<0.001);

[0028] Figure 3The effect of JDSJ on the average blood perfusion volume of SD rats with diabetic foot ulcer model (n=10); Note: Compared with the blank control group, # indicates a significant difference (P<0.05), ## indicates a significant difference (P<0.01), and ### indicates a significant difference (P<0.001); Compared with the model control group, * indicates a significant difference (P<0.05), ** indicates a significant difference (P<0.01), and *** indicates a significant difference (P<0.001);

[0029] Figure 4 The effect of JDSJ on VEGF content in SD rats with diabetic foot ulcer model (n=10); Note: Compared with the blank control group, # indicates significant difference (P<0.05), ## indicates significant difference (P<0.01), ### indicates significant difference (P<0.001); Compared with the model control group, * indicates significant difference (P<0.05), ** indicates significant difference (P<0.01), *** indicates significant difference (P<0.001);

[0030] Figure 5 The effect of JDSJ on the average optical density of VEGF in SD rats with diabetic foot ulcer model (n=10); Note: Compared with the blank control group, # indicates a significant difference (P<0.05), ## indicates a significant difference (P<0.01), and ### indicates a significant difference (P<0.001); compared with the model control group, * indicates a significant difference (P<0.05), ** indicates a significant difference (P<0.01), and *** indicates a significant difference (P<0.001). DETAILED DESCRIPTION

[0031] The present invention will be further described below by way of examples. It should be understood that the embodiments of the present invention are intended to illustrate the present invention rather than to limit the present invention. Simple modifications to the present invention according to the essence of the present invention all fall within the scope of protection claimed in the present invention. Unless otherwise indicated, the percentage of the amount of ethanol in the present invention is by volume, and v / v represents the volume ratio of the solution.

[0032] Example 1

[0033] The Chinese medicine composition for detoxification and tissue regeneration is made from the following raw materials:

[0034] 6kg of Lithospermum erythrorhizon, 6kg of Angelica sinensis, 2kg of Angelica dahurica, 2kg of Licorice, 1kg of Frankincense, and 1kg of Alum.

[0035] The preparation method of the active ingredient is as follows:

[0036] Take frankincense and alum and grind them into fine powder respectively, then sieve them;

[0037] Soak lithospermum officinale, angelica sinensis, angelica dahurica and licorice in 35 kg of tea oil for 5 days, stir once a day, and then boil at 105°C for 0.5 hours to obtain active ingredient oil;

[0038] Frankincense, Alum and Active Ingredient Oil are the active ingredients.

[0039] Example 2

[0040] The Chinese medicine composition for detoxification and tissue regeneration is made from the following raw materials:

[0041] 10kg of Lithospermum erythrorhizon, 10kg of Angelica sinensis, 6kg of Angelica dahurica, 6kg of Licorice, 4kg of Frankincense, and 5kg of Alum.

[0042] The preparation method of the active ingredient of the Chinese medicine composition for detoxification and tissue regeneration is as follows:

[0043] Take frankincense and alum and grind them into fine powder respectively, then sieve them;

[0044] Soak Lithospermum officinale, Chinese angelica, Angelica dahurica and Licorice in 100 kg of tea oil for 10 days, stir 3 times a day, and then boil at 110° C. for 2 hours to obtain active ingredient oil;

[0045] Frankincense, Alum and Active Ingredient Oil are the active ingredients.

[0046] Example 3

[0047] The Chinese medicine composition for detoxification and tissue regeneration is made from the following raw materials:

[0048] 8kg of Lithospermum erythrorhizon, 8kg of Angelica sinensis, 4kg of Angelica dahurica, 4kg of Licorice, 3kg of Frankincense, and 2kg of Alum.

[0049] The preparation method of the active ingredient of the Chinese medicine composition for detoxification and tissue regeneration is as follows:

[0050] Take frankincense and alum and grind them into fine powder respectively, then sieve them;

[0051] Soak lithospermum officinale, angelica sinensis, angelica dahurica and licorice in 50 kg of tea oil for 7 days, stir twice a day, and then boil at 108°C for 1 hour to obtain active ingredient oil;

[0052] Frankincense, Alum and Active Ingredient Oil are the active ingredients.

[0053] Example 4

[0054] The Chinese medicinal composition for detoxification and muscle regeneration is prepared from the following raw materials:

[0055] 7kg of Lithospermum officinale, 7kg of Angelica sinensis, 3kg of Angelica dahurica, 3kg of Licorice, 2kg of frankincense made with vinegar, and 3kg of alum.

[0056] The preparation method of the ointment comprises the following steps:

[0057] Soak lithospermum officinale, angelica, angelica dahurica and licorice in 40kg of tea oil for 6 days, stirring once a day, then simmer at 106℃ for 0.6 hours, add 7kg of beeswax to dissolve, filter, add frankincense and alum powder to the filtrate, mix well, and you have the product.

[0058] Example 5

[0059] The Chinese medicinal composition for detoxification and muscle regeneration is prepared from the following raw materials:

[0060] 9kg of Lithospermum officinale, 9kg of Angelica sinensis, 5kg of Angelica dahurica, 5kg of Licorice, 3kg of Vinegar-made Frankincense, and 4kg of Alum.

[0061] The preparation method of the active ingredient of the Chinese medicine composition for detoxification and tissue regeneration is as follows:

[0062] The preparation method of the ointment comprises the following steps:

[0063] Soak lithospermum officinale, angelica, angelica dahurica and licorice in 80kg of tea oil for 9 days, stirring 3 times a day, then decocted at 109℃ for 1.6 hours, add 10kg of white wax to dissolve, filter, add frankincense and alum powder to the filtrate, mix well, and you have the product.

[0064] Example 6

[0065] The Chinese medicinal composition for detoxification and muscle regeneration is prepared from the following raw materials:

[0066] 8kg of Lithospermum officinale, 8kg of Angelica sinensis, 4kg of Angelica dahurica, 4kg of Licorice, 2.66kg of frankincense made with vinegar, and 3.05kg of alum.

[0067] The preparation method of the ointment comprises the following steps:

[0068] Soak lithospermum, angelica, angelica dahurica and licorice in 50kg of tea oil for 7 days, stirring twice a day, then simmer at 106℃ for 1 hour, add 10kg of white wax to dissolve, filter, add frankincense and alum powder to the filtrate, mix well, and you have the product.

[0069] Example 7

[0070] The Chinese medicinal composition for detoxification and muscle regeneration is prepared from the following raw materials:

[0071] 7.5kg of Lithospermum officinale, 8.5kg of Angelica sinensis, 3kg of Angelica dahurica, 3kg of Licorice, 3.2kg of frankincense made with vinegar, and 2.5kg of alum.

[0072] The preparation method of the ointment comprises the following steps:

[0073] Soak lithospermum officinale, angelica, angelica dahurica and licorice in 70kg of tea oil for 7 days, stirring twice a day, then decocted at 106℃ for 1 hour, add 8kg of vaseline to dissolve, filter, add frankincense and alum powder to the filtrate, mix well, and you have the product.

[0074] Example 8

[0075] The Chinese medicinal composition for detoxification and muscle regeneration is prepared from the following raw materials:

[0076] 9kg of Lithospermum officinale, 9kg of Angelica sinensis, 4kg of Angelica dahurica, 5kg of Licorice, 3.5kg of Frankincense, and 2.5kg of Alum.

[0077] The preparation method of the ointment comprises the following steps:

[0078] Soak lithospermum officinale, angelica, angelica dahurica and licorice in 80kg of tea oil for 7 days, stir twice a day, then boil at 106℃ for 0.8 hour, add 9kg of lanolin and polyethylene glycol to dissolve, filter, add frankincense and alum powder to the filtrate, mix well, and you have the product.

[0079] Example 9

[0080] The Chinese medicinal composition for detoxification and muscle regeneration is prepared from the following raw materials:

[0081] 6.5kg of Lithospermum erythrorhizon, 5.5kg of Angelica sinensis, 3.5kg of Angelica dahurica, 5.5kg of Licorice, 3.5kg of Frankincense, and 1.5kg of Alum.

[0082] The preparation method of the ointment comprises the following steps:

[0083] Soak lithospermum officinale, angelica, angelica dahurica and licorice in 60kg of tea oil for 7 days, stirring twice a day, then simmer at 106℃ for 1.2 hours, add 12kg of beeswax, lanolin and vaseline and dissolve them, filter, add frankincense and alum powder to the filtrate, mix well, and you have the product.

[0084] Example 10

[0085] The Chinese medicinal composition for detoxification and muscle regeneration is prepared from the following raw materials:

[0086] 7kg of Lithospermum officinale, 7kg of Angelica sinensis, 5kg of Angelica dahurica, 5kg of Licorice, 1.8kg of frankincense made with vinegar, and 2kg of alum.

[0087] The preparation method of the ointment comprises the following steps:

[0088] Soak lithospermum officinale, angelica, angelica dahurica and licorice in 50kg of tea oil for 7 days, stirring twice a day, then simmer at 106℃ for 1.5 hours, add 11kg of beeswax, lanolin and vaseline and dissolve them, filter, add frankincense and alum powder to the filtrate, mix well, and you have the product.

[0089] Embodiment 11

[0090] The Chinese medicinal composition for detoxification and muscle regeneration is prepared from the following raw materials:

[0091] 8.5kg of Lithospermum erythrorhizon, 6.5kg of Angelica sinensis, 3.5kg of Angelica dahurica, 2.5kg of Licorice, 1.5kg of Frankincense, and 2.5kg of Alum.

[0092] The preparation method of the active ingredient is as follows:

[0093] Take frankincense and alum and grind them into fine powder respectively, then sieve them;

[0094] Soak lithospermum officinale, angelica sinensis, angelica dahurica and licorice in 60 kg of tea oil for 8 days, stir twice a day, and then boil at 110° C. for 2 hours to obtain active ingredient oil;

[0095] Frankincense, Alum and Active Ingredient Oil are the active ingredients.

[0096] The active ingredients are taken together with conventional film-forming materials and plasticizers to form a coating agent.

[0097] Example 12

[0098] The Chinese medicinal composition for detoxification and muscle regeneration is prepared from the following raw materials:

[0099] 7kg of Lithospermum officinale, 9kg of Angelica sinensis, 3kg of Angelica dahurica, 5kg of Licorice, 2kg of Vinegar-made Frankincense, and 4.5kg of Alum.

[0100] The preparation method of the active ingredient is as follows:

[0101] Take frankincense and alum and grind them into fine powder respectively, then sieve them;

[0102] Soak lithospermum officinale, angelica sinensis, angelica dahurica and licorice in 70 kg of tea oil for 7 days, stir once a day, and then boil at 103°C for 0.8 hours to obtain active ingredient oil;

[0103] Frankincense, Alum and Active Ingredient Oil are the active ingredients.

[0104] The active ingredients are taken together with conventional penetrants, adhesives, moisturizers and fillers to form a gel ointment.

[0105] Embodiment 13

[0106] Study on the therapeutic effect of Jiedu Shengji ointment on diabetic foot ulcer in rats

[0107] 1. Information on the test substance and reference substance

[0108] 1.1 Test article information

[0109] Name: Detoxification and muscle-regenerating cream, prepared by the method of Example 6

[0110] Code: JDSJ

[0111] Main ingredients: Lithospermum officinale, Angelica sinensis, Angelica dahurica, Licorice, Frankincense (made with vinegar), Alum

[0112] 1.2 Information on commercially available reference substances

[0113] Name: Commercially available detoxifying and regenerating ointment

[0114] Main ingredients: Lithospermum officinale, Angelica sinensis, Angelica dahurica, Licorice, Frankincense (made with vinegar), Calomel

[0115] 1.3 Negative Control Information

[0116] Name: Sodium Chloride Injection

[0117] 1.4 Positive Control Information

[0118] Name: Human epidermal growth factor gel

[0119] 2. Animal grouping

[0120] Group design: This experiment set up 5 groups, namely blank control group, model control group, test substance JDSJ group, commercially available detoxification and tissue regeneration cream group, and positive control group.

[0121] Number of animals: 10 per group, male.

[0122] Grouping method: Stratified random grouping was used for grouping. After the adaptive feeding was completed, 10 rats were randomly divided as the blank control group, and the remaining 70 rats were divided into the modeling group; after the modeling was successful, 40 SD rats with successful modeling were selected and randomly divided into 4 groups according to blood sugar and body weight, namely: blank control group, model control group, test substance JDSJ group, commercial detoxification and muscle-generating ointment group, and positive control group, 10 male rats in each group. The animal grouping is detailed in Table 2.

[0123] Table 2 Animal grouping

[0124]

[0125]

[0126] 3. Dosage design

[0127] 3.1 Information on Intended Clinical Use

[0128] Intended indications: promoting blood circulation and dispersing blood stasis, reducing swelling and relieving pain, removing toxins and pus, removing dead tissue and promoting tissue regeneration. Used for various wound infections and second-degree burns.

[0129] Intended Usage and Dosage: For external use, spread on gauze and apply to the affected area. The application thickness should be 1-2 mm, and the dosage is about 0.05g-0.1g per square centimeter.

[0130] Treatment course: Up to 21 days, a total of 1 course of treatment.

[0131] 3.2 Dosage design

[0132] The animal dosing area was the 3mm×7mm rectangular marked ulcer on the right dorsum of the foot. According to clinical information, the dosage per animal was set at 0.02g. Therefore, the positive control group of this experiment was given 0.02g of human epidermal growth factor gel, the test substance JDSJ was given 0.02g of JDSJ, and the commercially available detoxification and muscle regeneration cream group was given 0.02g of detoxification and muscle regeneration cream. The experimental dosage design is detailed in Table 3.

[0133] Table 3 Dosage design for the study of the therapeutic effect of JDSJ on diabetic foot ulcer model SD rats

[0134]

[0135] 7. Test methods

[0136] 4.1 Experimental modeling:

[0137] 4.1.1 Diabetes modeling method: A single intraperitoneal injection of 50 mg kg-1 of streptozotocin (STZ) solution was performed to establish a type 2 diabetes rat model. The blank control group was injected with an equal volume of citric acid-sodium citrate buffer.

[0138] 4.1.2 Screening of qualified rats: On the third day of modeling, blood was collected from the rat tail vein to test the blood glucose of the rats (blood glucose test strips), and rats without polydipsia, polyuria, weight loss and fasting blood glucose <16.7 mmol·mL-1 were eliminated.

[0139] 4.1.3 Diabetic foot ulcer modeling method: Rats were anesthetized by intraperitoneal injection of 50 mg kg-1 of sodium pentobarbital. A 3 mm × 7 mm rectangular mark was made on the dorsum of the right hind limb after disinfection with a stamp. The full-thickness skin was excised and the wound was disinfected with 0.5% iodine tincture.

[0140] 4.2 Administration site and route: Administer the drug to the skin of the ulcer on the dorsum of the right hind limb.

[0141] 4.3 Administration method: After the diabetic foot ulcer model is established, the corresponding drugs are administered according to Table 3 in 13.2. The drugs are directly applied to the foot ulcer, and then covered with a layer of sulfate paper and two layers of gauze, and then fixed with medical pressure-sensitive tape. The drugs are removed after about 5 hours of application, and the administration site is cleaned with 0.9% sodium chloride injection.

[0142] 4.4 Dosage: Unilateral dosage: 0.02 g / mouse.

[0143] 4.5 Frequency and duration of administration: Administer once a day for 18 consecutive days.

[0144] 4.6 The day of first administration is recorded as D 0 , the day before the first dose is D -1 , one day after the first dose is D 1 , and so on.

[0145] 5. Testing frequency and methods of various indicators

[0146] 5.1 Fasting blood glucose test in rats

[0147] D -1 , D 18 Collect venous blood samples from rats and use a biochemical analyzer to detect the fasting blood glucose values ​​of each group of animals (they were fasted for 12 hours the day before). 7 , D 14 The rats were fasted but not watered for 12 h, their tails were cut with a blade, and fasting blood glucose was measured with a blood glucose meter.

[0148] 5.2 Detection of wound healing rate in rats

[0149] D 0 , D 7 and D 14 The wound surface was covered with sulfuric acid paper to outline the area, and the wound was scanned and stored in JPG image format. The wound area was measured by ImageJ software, and the wound healing status was determined using the following formula: wound healing rate = (wound area after modeling - wound area after the drug administration cycle) / wound area after modeling × 100%.

[0150] 5.3 Local blood flow detection in rats

[0151] D 17 The laser speckle imaging system was used to detect the local average blood perfusion of the wound surface of each group of animals.

[0152] 5.4 Rat serum vascular endothelial growth factor (VEGF) levels

[0153] D 18 Blood was collected from the abdominal aorta and centrifuged at 3000 r / min for 10 min. The supernatant was taken and the VEGF content in the blood was determined by enzyme-linked immunosorbent assay. The detection operation was carried out strictly according to the instructions.

[0154] 5.5 Histopathological observation of rats

[0155] D 18 The wound healing tissues of animals in each group were taken and fixed in 4% polymethanol overnight. After dehydration, paraffin sections were prepared and stained with hematoxylin-eosin (HE) dye to observe tissue damage.

[0156] 5.6 Immunohistochemical detection of VEGF expression in rats

[0157] D 18 , 3 tissue wax blocks were cut into white slices from each group, dewaxed to water, antigen repaired, endogenous peroxidase blocked, blocked, primary and secondary antibodies added, DAB coloring (positive is brownish yellow), dehydrated, neutral gum sealed, etc., and observed and analyzed under an optical microscope after preparation. The pictures were analyzed using Image J software, the optical density value of positive cells per unit area was determined, and the average optical density value was calculated as the expression of the protein in each slice. The average optical density value is the ratio of the value (IOD) obtained by adding up the optical density values ​​of each point on the picture to the area (Area) in the corresponding target area.

[0158] 6 Data processing

[0159] The quantitative data generated in this experiment are expressed as mean ± standard deviation. MicroOffice Excel and SPSS21.0 software were used for data statistics, and Prism Graphpad8.0.2 software was used for drawing. One-way analysis of variance was used for comparison among multiple groups, and LSD-t test and Dunnett-t test were used for pairwise comparison among groups. P < 0.05 was considered statistically significant.

[0160] 7. Results Analysis

[0161] 7.1 Blood glucose in rats

[0162] Compared with the blank control group, the blood glucose level of the model control group increased, and the difference was significant (P<0.001). Compared with the model control group, there was no significant difference in the blood glucose levels of the drug-treated groups (P≥0.05). This shows that the model was successful and the blood glucose of the rats was maintained at a high level. Figure 1 , data see Table 3.

[0163] Table 3 Diabetic foot ulcer model SD rats D 7 Blood glucose (n=10)

[0164]

[0165] Note: Compared with the blank control group, # Indicates that the difference is significant (P<0.05). ## Indicates that the difference is significant (P<0.01). ### Indicates that the difference is significant (P<0.001); compared with the model control group, * Indicates that the difference is significant (P<0.05). ** Indicates that the difference is significant (P<0.01). ***Indicates that the difference is significant (P<0.001).

[0166] 7.2 Effect of JDSJ on wound healing in diabetic foot ulcer model SD rats

[0167] D7, compared with the blank control group, the wound healing rate of the model control group decreased, and the difference was significant (P<0.01); compared with the model control group, the healing rate of each drug-treated group increased, and the difference was not significant (P≥0.05). D14, compared with the blank control group, the wound healing rate of the model control group decreased, and the difference was significant (P<0.001); compared with the model control group, the healing rate of each drug-treated group increased, and the difference was significant (P<0.001). Figure 2 , data see Table 4.

[0168] Table 4 Effect of JDSJ on the healing rate of diabetic foot ulcer model SD rats (n=10, %)

[0169]

[0170] Note: Compared with the blank control group, # Indicates that the difference is significant (P<0.05). ## Indicates that the difference is significant (P<0.01). ### Indicates that the difference is significant (P<0.001); compared with the model control group, * Indicates that the difference is significant (P<0.05). ** Indicates that the difference is significant (P<0.01). *** Indicates that the difference is significant (P<0.001).

[0171] 7.3 Effect of JDSJ on the local mean blood perfusion in SD rats with diabetic foot ulcer model

[0172] Compared with the blank control group, the average blood perfusion volume in the model control group was significantly lower (P<0.05); compared with the model control group, the average blood perfusion volume in each drug-treated group had no significant difference (P≥0.05), but there was an upward trend. Figure 3 , data see Table 5.

[0173] Table 5 Effect of JDSJ on the mean blood perfusion volume of SD rats with diabetic foot ulcer model (n=5)

[0174]

[0175] Note: Compared with the blank control group, # Indicates that the difference is significant (P<0.05). ## Indicates that the difference is significant (P<0.01). ###Indicates that the difference is significant (P<0.001); compared with the model control group, * Indicates that the difference is significant (P<0.05). ** Indicates that the difference is significant (P<0.01). *** Indicates that the difference is significant (P<0.001).

[0176] 7.4 Effect of JDSJ on serum vascular endothelial growth factor (VEGF) content in SD rats with diabetic foot ulcer model

[0177] Compared with the blank control group, the VEGF content in the model control group had no significant difference (P ≥ 0.05), but there was a downward trend; compared with the model control group, the VEGF content in each drug-treated group had no significant difference (P ≥ 0.05), but there was an upward trend. Figure 4 , data see Appendix 6.

[0178] Table 6 Effect of JDSJ on VEGF levels in diabetic foot ulcer model SD rats (n=10)

[0179]

[0180] Note: Compared with the blank control group, # Indicates that the difference is significant (P<0.05). ## Indicates that the difference is significant (P<0.01). ### Indicates that the difference is significant (P<0.001); compared with the model control group, * Indicates that the difference is significant (P<0.05). ** Indicates that the difference is significant (P<0.01). *** Indicates that the difference is significant (P<0.001).

[0181] Effects of 7.5JDSJ on pathological changes of wound skin in SD rats with diabetic foot ulcer model

[0182] 7.5.1 Visual observation:

[0183] At the end of the medication, the skin of the blank control group was intact without obvious lesions. The skin ulcers on the dorsum of the right hind limb of the samples in the other groups healed to varying degrees, and scabs were seen on the wounds of some samples. In addition, no other abnormalities or lesions visible to the naked eye were found.

[0184] 7.5.2 HE staining results:

[0185] No abnormality was found in the skin tissue of the blank control group. Microscopic manifestations of post-traumatic repair were observed in the wounds of the other groups, mainly epidermal thickening to varying degrees, reduction of dermal appendages, focal inflammatory cell infiltration, granulation tissue, fibrous tissue hyperplasia and other lesions; the model control group had the most severe lesions, and compared with the model control group, each drug-treated group had fibrous tissue hyperplasia, and there was no significant difference in the degree of lesions among the drug-treated groups.

[0186] 7.5.3 Effect of JDSJ on the average optical density of VEGF expression in immunohistochemistry in SD rats with diabetic foot ulcer model

[0187] Compared with the blank control group, the average optical density of the rats in the model control group had no significant difference (P>0.05), but there was a downward trend; compared with the model control group, the average optical density of the rats in each drug-treated group had no significant difference (P>0.05), but there was an upward trend. Figure 5 , data see Appendix 7.

[0188] Table 7 Average optical density of VEGF expression in skin tissue of SD male rats

[0189]

[0190]

[0191] Note: Compared with the blank control group, # Indicates that the difference is significant (P<0.05). ## Indicates that the difference is significant (P<0.01). ### Indicates that the difference is significant (P<0.001); compared with the model control group, * Indicates that the difference is significant (P<0.05). ** Indicates that the difference is significant (P<0.01). *** Indicates that the difference is significant (P<0.001).

[0192] 8 Discussions

[0193] 8.1 Effect of JDSJ ointment on wound healing rate of diabetic foot ulcer in SD rats

[0194] JDSJ is composed of six herbs, namely, lithospermum officinale, angelica sinensis, angelica dahurica, liquorice, frankincense (made with vinegar), and alum, which are precisely matched according to the principle of monarch, minister, assistant, and envoy. It has stable and continuous chemical properties, controllable quality, and the combination of several herbs has the effects of detoxification, blood stasis removal, decomposition, and tissue regeneration. During the experimental observation period, the wounds of rats in each group gradually healed, and the wound surface was gradually covered with scabs of varying sizes. There were different degrees of swelling and exudation in the subscab space and around the wound, but there was no infection or suppuration. On the 7th day, the skin was red and swollen, granulation tissue sprouted on the wound, and the wound area had shrunk. Compared with the blank control group, the healing rate of the model control group was significantly reduced (P<0.01); compared with the model control group, the healing rates of the drug-treated groups were not significantly different (P≥0.05), but all increased. On the 14th day, except for the model control group, the wounds of the other groups healed well, and the wound area was significantly reduced. Compared with the blank control group, the healing rate of the model control group was significantly reduced (P<0.001); compared with the model control group, the healing rate of each drug-treated group was significantly increased (P<0.001). In summary, the tested detoxification and myogenicity can promote the growth of granulation tissue on the wound surface of rats with diabetic foot ulcers and accelerate the healing rate, which is consistent with the effect of commercially available detoxification and myogenicity and positive drugs.

[0195] 8.2 Effect of JDSJ ointment on VEGF and blood flow in SD rats with diabetic foot ulcer

[0196] Wound healing is affected by many factors. When tissue is damaged, wound healing is accompanied by the formation of granulation tissue. An important component of granulation tissue is new blood vessels, which supply oxygen to promote wound healing[1]. One of the early changes in diabetic vascular lesions is endothelial cell damage and dysfunction, which leads to slower blood circulation in the lower limbs. When diabetic rats are injured, endothelial cells proliferate actively, but cannot effectively form functional capillaries, which affects the blood oxygen supply to the wound and makes wound healing difficult[2]. Therefore, angiogenesis is crucial for wound healing of diabetic foot ulcers.

[0197] VEGF is the main growth factor that regulates blood vessels. It can trigger a series of repair reactions and effectively promote the formation of new blood vessels. Some studies have pointed out that the active period of VEGF expression is mainly concentrated in 4 to 8 days after the occurrence of wounds. In the process of wound healing, patients with diabetic foot ulcers show delayed VEGF expression, delayed angiogenesis, and impaired epidermal cell regeneration. Increasing VEGF levels can specifically cause vascular endothelial cell division and proliferation, increase vascular permeability, promote endothelial cell and monocyte migration, and induce angiogenesis [3]. Through vascular wall remodeling and fusion, functional collateral arterial circulation is gradually formed to rebuild the blood supply of ischemic tissues [4]. In this study, compared with the blank control group, the average blood perfusion volume in the model control group was significantly reduced (P < 0.05), the VEGF content showed a downward trend, and the VEGF expression showed a downward trend; compared with the model control group, the average blood perfusion volume, VEGF content, and VEGF expression of each drug-treated group showed an upward trend. Except for the blood flow in the model control group, there was no significant difference in the others (P ≥ 0.05). The reason may be that the active period of VEGF expression in the blank control group rats is mainly concentrated in 4-8 days after the wound occurs. At 18 days, the wound is basically healed, and the VEGF content will decrease slightly, but it is higher than that in the model control group. When there is relative oxygen deficiency around the blood vessels, VEGF transcription activation can be promoted, VEGF expression can be upregulated, and angiogenesis can be promoted [5-6]. The detoxification and muscle-generating ointment increases the level of VEGF, promotes cell migration and regeneration, and induces new blood vessel formation. The blood vessels generated in the later stage gradually improve the blood supply of the ulcer wound tissue, relieve tissue ischemia and hypoxia, and thus reduce VEGF expression. Therefore, compared with the model control group, there is no significant difference in VEGF expression in each drug-administered group (P≥0.05), but there is an upward trend. There is no significant difference between the drug-administered groups (P≥0.05). In summary, JDSJ can affect the level and expression of VEGF, and has an increasing trend on blood flow, which is basically consistent with the effects of commercially available detoxification and muscle-generating and positive drugs.

[0198] 8.3 Effect of JDSJ on wound pathology in SD rats with diabetic foot ulcer

[0199] The healing of ulcer wounds is affected by many factors. The wounds of diabetic foot ulcers are more difficult to heal and the inflammatory response is more severe. From the results of this experiment, it was found that compared with the blank control group, the model control group had fewer appendages, focal inflammatory infiltration, and bleeding. Compared with the model control group, the epidermis of each drug-treated group grew completely, the appendages increased, the focal inflammatory infiltration decreased, the bleeding decreased, and fibrous tissue hyperplasia appeared; there was no difference between the drug-treated groups. This shows that at the D18 stage, the wounds of each drug-treated group were repaired faster than those of the model control group, which can promote the healing of the wounds of diabetic foot ulcer rats.

[0200] 9 Conclusion

[0201] Under the conditions of this experiment, the test substance JDSJ can significantly improve the healing rate of diabetic foot ulcers in rats, affect the content and expression of VEGF, and increase the blood flow of rat wounds. There is no significant difference compared with the positive control group and the commercially available detoxification and muscle-generating ointment, indicating that the test substance JDSJ can promote the healing of diabetic foot ulcers in SD rats.

[0202] Although the present invention has been described in detail above by means of general description, specific implementation methods and tests, it is obvious to those skilled in the art that some modifications or improvements may be made to the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

[0203] References

[0204] [1]Noishiki C, Yuge S, Ando K, et al. Live imaging of angiogenesis during cutaneous wound healing in adult zebrafish[J].Angiogenesis, 2019; 22(2):341-54.

[0205] [2]Azevedo FM, Araújo-Filho I, Teixeira MMA, et al. Wound healing of diabetic rats treated with Moringa oleifera extract[J]. Acta Cir Bras, 2018; 33(9):799-805.

[0206] [3] Guo Jing, Meng Qinghai, Yin Qiuyi, et al. Study on the effect of Tongsaimai tablets on experimental diabetic foot model rats [J]. Chinese Journal of Traditional Chinese Medicine, 2014, 39(11): 2091-2096.

[0207] [4] Tian Fei. Study on the correlation between HIF-1α, VEGF and MVD and psoriasis[D]. Shijiazhuang: Hebei Medical University, 2008.

[0208] [5] Zhu Qiuxiao, Hao Huiyao, Liu Zibo, et al. Effects of rheumatoid arthritis on vascular endothelial cell injury in diabetic rats with macroangiopathy by regulating HIF-1α / VEGF pathway[J]. Chinese Journal of Clinical Pharmacology, 2024, 40(06): 859-863.

[0209] [6] Chen Shengye, Gan Yu. Effect of Shengji Yuhong ointment on the expression of bFGF, HIF-1α and VEGF in diabetic foot ulcer tissues of rats [J]. Western Chinese Medicine, 2021, 34(09): 23-26.

Claims

1. Use of a detoxifying and myogenic composition in the preparation of a drug for treating diabetic foot ulcers, characterized in that: The detoxification and muscle-building composition is made of the following raw materials in parts by weight: 60-100 parts of lithospermum, 60-100 parts of angelica, 20-60 parts of angelica dahurica, 20-60 parts of liquorice, 10-40 parts of frankincense and 10-50 parts of alum.

2. The use according to claim 1, characterized in that Made from the following raw materials in parts by weight: 70-90 parts of lithospermum officinale, 70-90 parts of angelica sinensis, 30-50 parts of angelica dahurica, 30-50 parts of liquorice, 20-30 parts of vinegar-processed frankincense, and 30-40 parts of alum.

3. The use according to claim 1 or 2, characterized in that The preparation method of the active ingredient comprises the following steps: Take frankincense and alum and grind them into fine powder respectively, then sieve them; Soaking lithospermum officinale, angelica sinensis, angelica dahurica and licorice in 350-1000 parts by weight of tea oil for 5-10 days, stirring 1-3 times a day, and then boiling at 105-110° C. for 0.5-2 hours to obtain active ingredient oil; Frankincense, Alum and Active Ingredient Oil are the active ingredients.

4. The use according to claim 3, characterized in that The preparation method of the active ingredient comprises the following steps: Take frankincense and alum and grind them into fine powder respectively, then sieve them; Soaking lithospermum officinale, angelica sinensis, angelica dahurica and licorice in 400-700 parts by weight of tea oil for 5-10 days, stirring 1-3 times a day, and then boiling at 105-110° C. for 0.5-2 hours to obtain active ingredient oil; Frankincense, Alum and Active Ingredient Oil are the active ingredients.

5. The use according to claim 1 or 2, characterized in that The preparation method of the ointment comprises the following steps: Soak lithospermum officinale, angelica sinensis, angelica dahurica and licorice in 350-1000 parts by weight of tea oil for 5-10 days, stir 1-3 times a day, then boil at 105-110° C. for 0.5-2 hours, add 60-200 parts by weight of matrix to dissolve, filter, add frankincense and alum powder to the filtrate, mix well, and obtain; The matrix is ​​one or a combination of beeswax, lanolin, vaseline, polyethylene glycol and white wax.

6. The use according to claim 5, characterized in that The preparation method of the ointment comprises the following steps: Soak lithospermum officinale, angelica sinensis, angelica dahurica and licorice in 400-700 parts by weight of tea oil for 5-10 days, stir 1-3 times a day, then boil at 105-110° C. for 0.5-2 hours, add 80-120 parts by weight of matrix to dissolve, filter, add frankincense and alum powder to the filtrate, mix well, and obtain; The matrix is ​​one or a combination of beeswax, lanolin, vaseline, polyethylene glycol and white wax.

7. A Chinese medicine preparation for treating diabetic foot ulcer, prepared by combining the detoxifying and myogenic composition according to any one of claims 1 to 6 with a pharmaceutically acceptable carrier.

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