Application of arbutin in preparation of medicine for promoting wound healing of diabetic foot ulcer
By preparing a topical cream using bear-like arbutin, the production of inflammatory cytokines and the activation of fibroblasts are inhibited, thus solving the problem of delayed healing of diabetic ulcers and significantly accelerating the healing and tissue regeneration of diabetic foot ulcers.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-02-02
- Publication Date
- 2026-03-27
AI Technical Summary
The lack of effective drugs in current technology to promote the healing of diabetic ulcer wounds, especially the delayed healing and poor healing of diabetic foot ulcers, has become a major global public health challenge.
A topical cream made from bearberry extract promotes the healing of diabetic foot ulcers by inhibiting the production of inflammatory cytokines and activating fibroblasts.
It significantly accelerates the healing rate of ischemic foot ulcers in diabetic rats, reduces inflammatory cell infiltration, promotes the growth of regenerating epithelium in the wound, increases collagen content, improves collagen fiber distribution, enhances wound tissue structure, activates fibroblast migration ability, and promotes tissue regeneration and high-quality healing.
Smart Images
Figure CN121731284A_ABST
Abstract
Description
Technical Field
[0001] This application relates to the field of biomedical technology, and in particular to the application of arbutin in the preparation of drugs that promote the healing of diabetic foot ulcers. Background Technology
[0002] Diabetic patients' skin is prone to damage, with diverse mechanisms leading to delayed or non-healing wounds, a challenging and pressing issue in clinical practice. As of 2018, the global incidence of diabetic skin ulcers was as high as 20%. Diabetic ulcers have become a leading cause of chronic, non-healing wounds. Among these, diabetic foot ulcers, with their high incidence, poor healing, and high disability rate, have become a major challenge in global public health. The core mechanism behind the difficulty in healing diabetic ulcers lies in the multi-system damage caused by hyperglycemia: microvascular complications lead to tissue ischemia and hypoxia, neuropathy delays the perception of damage, immunodeficiency increases the risk of infection, the accumulation of advanced glycation end products (AGEs) damages collagen structure, and abnormal fibroblast function delays tissue regeneration. These factors collectively lead to chronic inflammation, delayed repair, and recurrent infections. Summary of the Invention
[0003] The purpose of this application is to address the technical problem of the lack of drugs for diabetic ulcers in the prior art.
[0004] To achieve the above objectives, this application provides the following technical solution:
[0005] Application of arbutin in the preparation of drugs that promote the healing of diabetic foot ulcers.
[0006] Preferably, the arbutin promotes the healing of diabetic foot ulcers by inhibiting the production of inflammatory cytokines.
[0007] Preferably, the bearberry extract promotes the healing of diabetic foot ulcers by activating fibroblasts.
[0008] This application also provides a drug for promoting the healing of diabetic foot ulcers, the drug containing arbutin.
[0009] Preferably, the drug is a topical drug, and the drug also includes other medically acceptable excipients.
[0010] Preferably, the drug is a cream containing glycerin.
[0011] Preferably, the drug preparation method is as follows:
[0012] Add an appropriate amount of glycerin to bear bamboo extract, stir to mix, and let stand.
[0013] Weigh stearic acid, glyceryl monostearate, light liquid paraffin, polysorbate 80, sorbitan oleate, ethylparaben, and benzoic acid, mix them well, place them in a container, and heat them to 78℃-82℃ until they are completely melted. After standing and cooling, add the arbutin-glycerin mixture, mix thoroughly, and obtain arbutin cream, which is then packaged and stored.
[0014] Preferably, the bearberry extract cream contains 0.00625g bearberry extract / g cream.
[0015] Compared with the prior art, this application has the following beneficial effects:
[0016] This application verifies that arbutin can significantly accelerate the healing rate of ischemic foot ulcers in diabetic rats by inhibiting the production of inflammatory cytokines, reducing erythema, edema, and inflammatory exudation in diabetic foot ulcers, preventing clinical infection, and creating a favorable healing environment for the wound.
[0017] In addition, bearberry extract can reduce inflammatory cell infiltration in the wound area, promote the regular and more complete growth of regenerated epithelium in the wound, and promote the formation of hair follicles around the wound, significantly improving the wound re-epithelialization rate and accelerating the maturation and formation of granulation tissue.
[0018] Furthermore, bearberry extract can increase the total amount of collagen in the wound site, improve the uneven distribution of collagen fibers, make the fibers more neat, dense and thick, enhance the tensile strength and structural support of the wound tissue, provide a stable structural foundation for wound repair, and help the wound heal with high quality.
[0019] By activating the physiological activity of fibroblasts, bearberry extract significantly promotes the migration ability of human foreskin fibroblasts (HFF-1) under normal culture conditions, delivering sufficient functional cells for wound healing, accelerating tissue regeneration, and promoting the wound repair process.
[0020] Bearberry extract can also reverse the damage to the migration ability of fibroblasts caused by a high-sugar, high-fat environment, improve the problem of abnormal cell function in the pathological state of diabetes, solve the core problem of delayed wound healing at the cellular level, and effectively improve the poor healing of diabetic foot ulcers. Attached Figure Description
[0021] Figure 1 The chemical structural formula of arbutin is shown.
[0022] Figure 2 The healing status of foot ulcers in diabetic rats; A: Representative photographs of wound healing in each group; B: Statistical chart of wound healing rate in each group. **** P<0.0001 vs DFU.
[0023] Figure 3HE and Masson staining of skin tissue from foot wounds in diabetic rats; A: Representative HE-stained images and magnified views of wounds in each group on day 21. B: Statistical chart depicting re-epithelialization of wounds in each group on day 21 (n=3). C: Representative Masson stained images and magnified views of wounds in each group on day 21. D: Statistical chart of collagen content in wounds in each group on day 21 (n=3). **P < 0.01 vs DFU
[0024] Figure 4 The effects of arbutin on fibroblast migration ability: A: Morphological representation of scratch healing area of HFF-1 cells under different treatment conditions; B: Representative representation of Transwell-migrated crystal violet-stained cells of HFF-1 cells under different treatment conditions; C: Quantitative analysis of scratch assay; D: Statistical graph of Transwell-migrated crystal violet-stained cell area. * P<0.05, ** P<0.01, *** P<0.001. Detailed Implementation
[0025] This application discloses the use of arbutin in the preparation of a drug to promote the healing of diabetic foot ulcers, the chemical structural formula of which is as follows: Figure 1 As shown, the bearberry extract promotes the healing of diabetic foot ulcers by inhibiting the production of inflammatory cytokines and activating fibroblasts.
[0026] This application also provides a drug for promoting the healing of diabetic foot ulcers, the drug containing arbutin. The drug is a topical application.
[0027] Preferably, the drug is a cream containing glycerin, and the cream is prepared as follows:
[0028] Add an appropriate amount of glycerin to arbutin, stir and mix, then let stand. Weigh stearic acid, glyceryl monostearate, light liquid paraffin, polysorbate 80, sorbitan oleate, ethylparaben, and benzoic acid, mix well, place in a container and heat to 78℃-82℃ until completely melted. After cooling, add the arbutin-glycerin mixture and mix thoroughly to obtain arbutin cream (0.00625g arbutin / g cream), which is then packaged and stored.
[0029] The above content will be explained in conjunction with specific verification experiments:
[0030] I. Experimental Procedure:
[0031] 1. Preparation of Bear Bamboo Extract Ointment
[0032] Arbutin (Shanghai Yuanmu Biotechnology Co., Ltd.) was mixed with an appropriate amount of glycerin and allowed to stand. The raw materials were weighed according to the ratio, and the components of the cream were heated to 78°C-82°C to completely melt them. After standing and cooling, the arbutin-glycerin mixture was added and mixed thoroughly to obtain the arbutin cream. 625mg of arbutin was added to 100g of the cream. The arbutin cream was then packaged and stored.
[0033] The mass ratios of the components in the cream are shown below:
[0034]
[0035] 2. Animal experiments
[0036] The male SD rats (weighing 180±20 g) used in this study were obtained from the Model Animal Research Center of Nantong University, and the animal experiments have been ethically approved (No.: S20240718-101). They were kept in an environment of 22-25℃, following a 12-hour light-dark cycle, provided with standard animal feed, and had access to tap water at all times.
[0037] Animal models, grouping, and drug administration
[0038] Forty-five male SD rats weighing 180±20 grams were randomly divided into three groups according to their weight: normal control group (NC), diabetic foot ulcer model control group (DFU), and arbutin treatment group (Jaranol), with 15 rats in each group.
[0039] After a week of acclimatization, the model was established. Rats were fasted but allowed to drink water for 10 hours. Mice in the DFU and arbutin groups were intraperitoneally injected with STZ-citrate buffer at a dose of 65 mg / kg, while the control group was injected with citrate buffer. After injection, blood was collected from the tail tip to measure random blood glucose levels for three consecutive days. Rats with persistent blood glucose levels above 16.7 mmol / L and obvious symptoms of polyuria, polydipsia, polyphagia, and weight loss were considered to have successfully established the model.
[0040] In all groups, blood glucose levels were monitored daily using a Johnson & Johnson OneTouch Ultra glucometer. After one month, rats with blood glucose levels above 16.65 mmol / L were used for subsequent experiments (type 1 diabetic rats). Rats that did not meet the blood glucose criteria were discarded. T1DM rats with implanted insulin pumps continued to be fed for 4 weeks to simulate the late stage of diabetes. After anesthesia with 1% pentobarbital at 0.05 mg / kg, the rats were placed in a supine position and secured, and the skin was disinfected. A skin incision was made along the right femoral artery, and the femoral vein and artery were carefully dissected. The proximal and distal ends were ligated and cut, and the skin was sutured. After disinfection with povidone-iodine, a 0.8 cm diameter circular hole was made in the rat's foot using a sterile bio-sampling needle, and the size of the ulcer wound was photographed and recorded. After surgery, the foot wounds of each group of rats were treated daily with either a control agent or a purpuric acid cream. After the surgery, each rat was placed in a separate cage to prevent bites.
[0041] 3. Drug use and collection of damaged tissues
[0042] The group receiving the bear-root extract cream used 0.5 g / cm² / day of bear-root extract cream daily at 9:00 AM. The DFU control group received the pharmaceutical excipients contained in the bear-root extract cream concurrently. Administration was once daily. Tissue samples were retrieved from the wound via euthanasia 14 or 21 days after the end of treatment.
[0043] 4. Hematoxylin-eosin (HE) staining and Masson staining
[0044] Full-thickness wound tissue samples were carefully collected from the damaged area. Specimens were fixed overnight in 4% paraformaldehyde solution. Subsequently, the samples were embedded, and the embedded tissue was prepared into 8-micrometer-thick tissue sections. HE staining (product number: C0105S; Beyotime, China) and Masson's trichrome staining (product number: C0189S; Beyotime, China) were performed according to the manufacturer's instructions using staining kits. The level of re-epithelialization and collagen content in the skin areas stained with HE and Masson's trichrome were determined using ImageJ software analysis.
[0045] 5. Wound healing rate
[0046] The wound was photographed on days 3, 7, and 14, and the wound area was analyzed using ImageJ software. The wound healing rate (WHR) was calculated using the following formula: WHR = (OA - CA) / OA × 100%
[0047] In this context, OA and CA represent the original area and current area of the wound, respectively.
[0048] 6. Culture of human foreskin fibroblasts
[0049] Human foreskin fibroblasts (HFF-1 cells, ATCC cell bank) were cultured in low-glucose DMEM medium (Gibco, Thermo Fisher Scientific) supplemented with 10% fetal bovine serum (Gibco, Thermo Fisher Scientific) and complete medium supplemented with 1% PS (Gibco, Thermo Fisher Scientific) at 37°C in a humid environment containing 5% carbon dioxide.
[0050] 7. Transwell experiment
[0051] Cell migration and invasion were assessed using Transwell chambers (MilliporeSigma) with an 8-micron pore size. 4 × 10⁴ cells were placed in each chamber. 4 Cells suspended in serum-free medium were seeded into the upper chamber, while the lower chamber contained complete medium with 10% fetal bovine serum. After incubation at 37°C for 24 hours, the cells were fixed with 4% paraformaldehyde, stained with crystal violet, and any non-migrating cells were removed. Migrating and invasive cells were counted in randomly selected fields of view.
[0052] 8. Scratch test
[0053] Cell migration was assessed using a scratch assay. HFF-1 cells were sputtered at a rate of 1 × 10⁶ cells / year. 5 Cells were seeded at a density of [number] cells / mL in 6-well plates to achieve approximately 95% confluence. Four parallel scratches were made in each well using a 200 μL pipette tip. Exfoliated cells and debris were removed by rinsing with phosphate-buffered saline (PFS). Cells were then subjected to various treatments, including 10 μg / μL of Exos or no treatment (control condition). Cells were then cultured for 0, 4, 12, or 24 hours. The width of the scratches was visually observed using an optical microscope (Leica, Germany) and measured using ImageJ software.
[0054] 9. Statistical Analysis
[0055] The experimental data were statistically analyzed using GraphPadPrism software. The t-test was used to compare two groups of data, and the one-way ANOVA was used to compare multiple groups. P < 0.05 was considered statistically significant.
[0056] II. Experimental Conclusions:
[0057] 1. Bear bamboo extract cream can significantly promote the healing of diabetic foot wounds.
[0058] This application compares the healing efficiency of the diabetic foot ulcer group (DFU group) and the bearberry extract cream group during the wound repair process.
[0059] The results showed that the DFU group exhibited inflammatory erythema and swelling at the wound site in the early stages due to reduced tissue perfusion. However, the cream group significantly reduced erythema, edema, and inflammatory exudation at the wound site, with no signs of clinical infection observed, and wound healing was accelerated. Figure 2 (AB). These results indicate that the bearberry extract cream can effectively promote the healing of ischemic foot ulcers in diabetic rats.
[0060] 2. Bear-Bamboo Extract Cream can significantly promote the formation of granulation tissue in diabetic foot wounds.
[0061] HE staining is an important histological indicator for assessing the pathological progress of wound healing. To further assess the progress of wound healing, wound tissue was collected on day 21 and histologically analyzed using HE staining.
[0062] The results showed that, compared with the DFU group, the regenerated epithelium in the rat paw wound of the bear-like bamboo extract cream group was more intact, with regular layers, no obvious inflammatory cell infiltration, and hair follicles began to form around the wound. The re-epithelialization rate of the DFU group was 61.5%, while the re-epithelialization rate of the bear-like bamboo extract cream group (82.4%) was higher than that of the DFU group. Figure 3 (AB). These results indicate that bearberry extract cream can effectively reduce inflammatory infiltration in the wound area of diabetic rats and promote reepithelialization of wound tissue, thereby accelerating wound healing.
[0063] Collagen deposition and subsequent remodeling play a crucial role in enhancing the tensile strength of tissues. This process also generates mechanical tension within the skin and scar tissue, thereby promoting wound healing by providing structural support. To further investigate collagen deposition at the wound site, Masson's trichrome staining was performed on skin tissue sections from the rat paw wound area. On day 21, the DFU group showed uneven collagen fiber distribution, low content (60.4%), and areas of inflammation. Compared to the DFU group, the collagen content of the bear-root extract cream was significantly increased (84.1%), with fibers arranged neatly and densely. Figure 3 These findings indicate that bearberry extract cream can effectively reduce inflammatory infiltration in wound tissues of diabetic rats and promote collagen deposition, thereby accelerating wound healing.
[0064] 3. Bearberry extract cream significantly promotes the migration ability of HFF-1 cells in normal and high-sugar, high-fat cultures.
[0065] This application investigated the effect of arbutin on the migration ability of HFF-1 cells using a scratch assay. Images were acquired at 0h, 12h, and 24h after scratching. The results showed that arbutin significantly promoted the migration of normally cultured HFF-1 cells. The blank area in the HG+PA group was much larger than that in the Nor group, while the blank area in the arbutin group was not significantly different from that in the Nor group. This indicates that arbutin can partially reverse the damage to cell migration ability caused by high glucose and high lipid intake. Figure 4 A, 4C).
[0066] In addition, this study further verified the above results using Transwell experiments, and the results showed that ( Figure 4 (B, 4D) Arbutin significantly increased HFF-1 migration. This suggests that arbutin can effectively reverse the damage to cell migration ability caused by high sugar and high lipid levels, which is consistent with the results of the scratch assay.
[0067] This application systematically evaluated the therapeutic effect of topical bearberry extract cream on the healing of diabetic foot ulcers through animal and cell experiments, and explored its mechanism of action. A rat model of diabetic foot ulcers was established, and rats were divided into groups for drug treatment. Simultaneously, related experiments were conducted on human foreskin fibroblasts. Through various detection methods and statistical analysis, the positive effects of bearberry extract cream on promoting wound healing, granulation tissue formation, collagen deposition, and fibroblast migration were confirmed.
Claims
1. Application of arbutin in the preparation of drugs that promote the healing of diabetic foot ulcers.
2. The application of arbutin according to claim 1 in the preparation of a drug for promoting the healing of diabetic foot ulcers, characterized in that: The arbutin promotes the healing of diabetic foot ulcers by inhibiting the production of inflammatory cytokines.
3. The application of arbutin according to claim 1 in the preparation of a drug for promoting the healing of diabetic foot ulcers, characterized in that: The arbutin promotes the healing of diabetic foot ulcers by activating fibroblasts.
4. A drug for promoting the healing of diabetic foot ulcers, characterized in that: The drug contains bearberry extract.
5. The drug for promoting the healing of diabetic foot ulcers according to claim 4, characterized in that: The medicine is a topical medicine, and it also includes other medically acceptable excipients.
6. The drug for promoting the healing of diabetic foot ulcers according to claim 5, characterized in that: The drug is a cream containing glycerin.
7. A drug for promoting the healing of diabetic foot ulcers according to claim 6, characterized in that: The drug preparation method is as follows: Add an appropriate amount of glycerin to bear bamboo extract, stir to mix, and let stand. Weigh stearic acid, glyceryl monostearate, light liquid paraffin, polysorbate 80, sorbitan oleate, ethylparaben, and benzoic acid, mix them well, place them in a container, and heat them to 78℃-82℃ until they are completely melted. After standing and cooling, add the arbutin-glycerin mixture, mix thoroughly, and obtain arbutin cream, which is then packaged and stored.
8. A drug for promoting the healing of diabetic foot ulcers according to claim 7, characterized in that: The bear-bamboo extract cream contains 0.00625g bear-bamboo extract / g cream.