Application of dehydrobulitol in preparation of medicine for improving or promoting wound healing

Abstract

The invention discloses an application of dehydropalmitol in preparation of a medicine for improving or promoting wound healing, and belongs to the field of biological medicine. In the invention, the improvement or promotion of wound healing comprises the increase of the length of regenerated epidermis, the increase of collagen area, the promotion of wound vascularization, the increase of capillary density and / or the increase of wound healing rate. According to the present invention, the research results show that the dehydrobulitol can effectively improve or promote the wound healing, has advantages of simpleness, safety, reliability, high efficiency and the like, and has wide application prospects in the wound healing field.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, and in particular relates to the application of dehydroepiandrosterol in the preparation of drugs that improve or promote wound healing. Background Technology

[0002] Wound healing is a complex process involving coagulation, inflammation, removal of damaged matrix components, and subsequent stages such as cell proliferation and migration, angiogenesis, matrix synthesis and deposition, reepithelialization, and remodeling. Wounds, such as chronic wounds, significantly impact patients' quality of life and mental and physical health.

[0003] Treatment methods for wounds mainly include traditional wound dressings, hydrogel wound dressings, antibacterial dressings, stem cell therapy, and traditional transdermal drug delivery. However, these methods still have problems such as poor efficacy, lack of corresponding biological functions, difficulty in changing wound dressings, inability to effectively protect wounds, easy to cause local oxidative stress in wounds, vascular damage and secondary infection, limited biochemical functions, inability to achieve precise delivery of active substances, and long treatment cycles.

[0004] Therefore, there is an urgent need for a simple, safe, reliable, and effective drug to promote wound healing. Summary of the Invention

[0005] To address at least some of the technical problems in the prior art, the present invention provides the use of dehydroepiandrosterol in the preparation of medicaments for improving or promoting wound healing. Specifically, the present invention includes the following.

[0006] A first aspect of the present invention provides the use of dehydroepiandrosterol in the preparation of a medicament for improving or promoting wound healing.

[0007] In some embodiments, the use of dehydroepiandrosterol according to the first aspect of the invention in the preparation of a medicament for improving or promoting wound healing, wherein the improvement or promotion of wound healing comprises at least one of the following: (1) Increase the length of the regenerated epidermis; (2) Increase collagen surface area; (3) Promotes wound vascularization; (4) Increase capillary density; (5) Increase the content of α-SMA or CD31.

[0008] In some embodiments, the use of dehydroepiandrosterol according to the first aspect of the invention in the preparation of a medicament for improving or promoting wound healing, wherein the wound includes a skin wound.

[0009] In some embodiments, the use of dehydroepiandrosterol according to the first aspect of the invention in the preparation of a medicament for improving or promoting wound healing, wherein the wound includes acute wounds and chronic wounds.

[0010] In some embodiments, the use of dehydroepiandrosterol according to the first aspect of the invention in the preparation of a medicament for improving or promoting wound healing, wherein the acute wound includes at least one of abrasions, lacerations, cuts, punctures, and burns, and the chronic wound includes diabetic wounds.

[0011] A second aspect of the present invention provides a method for preparing dehydroepiandrosterol, comprising the following steps: (1) Extract Sedum sarmentosum with water to obtain Sedum sarmentosum water extract; (2) Extract the aqueous extract of Sedum sarmentosum using an organic solvent to obtain Sedum sarmentosum extract; (3) The extract of Sedum sarmentosum is purified and separated to obtain the dehydroepiandrosterol.

[0012] In some embodiments, according to the second aspect of the present invention, the method for preparing dehydroepiandrosterol is wherein, in step (1), the material-to-liquid ratio is 1:(5-50).

[0013] In some embodiments, according to the method for preparing dehydroepiandrosterol according to the second aspect of the present invention, in step (1), the water extraction time is 0.5-5 h.

[0014] In some embodiments, according to the method for preparing dehydroepiandrosterol according to the second aspect of the present invention, the organic solvent includes at least one selected from petroleum ether, chloroform, ethyl acetate, n-butanol, ethanol, acetone and cyclohexane.

[0015] A third aspect of the present invention provides a method for increasing the content of α-SMA and / or CD31 in cells in vitro, comprising the step of contacting cells with dehydroepiandrosterone.

[0016] The present invention has found that dehydroepiandrosterone can effectively improve or promote wound healing, and has the advantages of being simple, safe, reliable and efficient, and has broad application prospects in the field of wound healing, especially chronic wound healing. Attached Figure Description

[0017] Figure 1 The changes in blood glucose levels in mice during the construction of the animal model are shown.

[0018] Figure 2 The effects of different Sedum sarmentosum extracts on wound healing in animals were demonstrated.

[0019] Figure 3The effects of different Sedum sarmentosum extracts on wound healing in animals were demonstrated.

[0020] Figure 4 The quantitative analysis results of different Sedum sarmentosum extracts on wound healing in animals are shown.

[0021] Figure 5 The quantitative analysis results of different Sedum sarmentosum extracts in cell experiments are shown. In the figure, A and B are cell viability and cell migration rate of HUVEC cells, respectively; C and D are cell migration rate and cell viability of HaCat cells.

[0022] Figure 6 The effects of different polar components on cell proliferation in cell experiments were demonstrated.

[0023] Figure 7 The results of high performance liquid chromatography analysis of the components in tubes 31-43 are shown.

[0024] Figure 8 The results of high-performance liquid chromatography analysis of the target product are shown.

[0025] Figure 9 The effects of the target product in cell experiments are shown, where A represents the experimental results in HUVEC cells; and B and C represent the experimental results in HaCat cells.

[0026] Figure 10 The target compound is shown. 1 H-NMR analysis results.

[0027] Figure 11 The target compound is shown. 13 C-NMR analysis results.

[0028] Figure 12 The effects of different doses of dehydroepiandrosterol on promoting wound healing in animals were shown, where A represents the qualitative analysis results and B represents the quantitative analysis results.

[0029] Figure 13 The results of a study on the mechanism of different doses of dehydroemetrol on wound healing in animals are shown. Among them, A is the result of immunohistochemical analysis; B is the effect of different doses of dehydroemetrol on the length of regenerated epidermis; C is the effect of different doses of dehydroemetrol on Col collagen; D is the effect of different doses of dehydroemetrol on the α-SMA vascular area; and E is the effect of different doses of dehydroemetrol on capillary density. Detailed Implementation

[0030] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0031] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that the upper and lower limits of the range and each intermediate value between them are specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, are also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0032] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0033] application One aspect of the present invention provides the use of dehydroepiandrosterol in the preparation of a medicament for improving or promoting wound healing. Dehydroepiandrosterol can be obtained through chemical synthesis or plant extraction, examples of which include, but are not limited to, *Sedum sarmentosum*, *Sedum morganianum*, *Bambusa textilis*, *Pteris vittata*, *Morinda officinalis*, *Bidens pilosa*, *Artemisia annua*, and *Polygonum multiflorum*. The dehydroepiandrosterol has a molecular weight of 222 and a molecular formula of C2. 13 H 18 O3, chemical structural formula as shown in Formula I: Formula I.

[0034] In a preferred embodiment, the improvement or promotion of wound healing includes, but is not limited to, increasing the length of regenerated epidermis; increasing collagen surface area; promoting wound vascularization; increasing capillary density; increasing the content of α-SMA or CD31; and improving wound healing rate. The degree of improvement or promotion is at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, or 100%.

[0035] In this invention, the wound includes, but is not limited to, skin wounds. The wound includes acute wounds and chronic wounds. Acute wounds include, but are not limited to, abrasions, lacerations, cuts, punctures, and burns. Chronic wounds include, but are not limited to, diabetic wounds. In a preferred embodiment, the chronic wound is a diabetic wound.

[0036] The medicament of this invention can be formulated into various clinically acceptable dosage forms, including but not limited to solutions, tinctures, ointments, creams, gels, powders, aerosols, sprays, patches, transdermal patches, tablets, capsules, granules, suspensions, emulsions, syrups, and injections. There are no particular limitations on the method of administration, but representative methods include, but are not limited to, topical administration, oral administration, and injection.

[0037] In this invention, the prevention, improvement, or treatment is achieved by administering a therapeutically effective amount of the drug to a subject in need. Subjects include, but are not limited to, mammals, including but not limited to, humans, mice, rabbits, cats, dogs, cattle, sheep, and pigs.

[0038] The therapeutically effective dose described in this invention refers to a pharmaceutically recognized effective dosage, meaning that the amount of the drug in this invention is sufficient to significantly promote wound healing without causing serious side effects. The daily dosage of the active pharmaceutical ingredient (e.g., dehydroepiandrosterol) of this invention is typically 0.01-500 mg, preferably 1-500 mg, more preferably 10-400 mg, further preferably 30-300 mg, and even more preferably 60-300 mg, for example 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg, 170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg, 205 mg, 210 mg, 215 mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, etc. 245 mg, 250 mg, 255 mg, 260 mg, 265 mg, 270 mg, 275 mg, 280 mg, 285 mg, 290 mg, 295 mg, 300 mg, or any dose within the above range. It can be administered as a single dose once daily, divided into multiple doses daily, or at intervals.

[0039] Preparation method One aspect of the present invention provides a method for preparing dehydroepiandrosterol, comprising the following steps: (1) Extract Sedum sarmentosum with water to obtain Sedum sarmentosum water extract; (2) Extract the aqueous extract of Sedum sarmentosum using an organic solvent to obtain Sedum sarmentosum extract. Examples of organic solvents include, but are not limited to, petroleum ether, chloroform, ethyl acetate, n-butanol, ethanol, acetone, cyclohexane, etc. (3) The extract of Sedum sarmentosum is purified and separated to obtain the dehydroepiandrosterol.

[0040] In a preferred embodiment, the preparation method of the present invention includes the following steps: (1) Water extraction of *Sedum sarmentosum* is performed at 50-100℃, preferably 55-100℃, even more preferably 60-100℃, further preferably 65-100℃, and more preferably 70-100℃, for example 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, or 100℃ for 0.5-5 h, preferably 0.5-4.5 h, even more preferably 0.5-4 h, further preferably 0.5-3.5 h, and more preferably 1-3 h, for example 1, 1.5, 2, 2.5, or 3 h. h, the material-to-liquid ratio is 1:(5-50), preferably 1:(5-45), even more preferably 1:(5-40), more preferably 1:(5-30), for example 1:5, 1:10, 1:15, 1:20, 1:25, 1:30, to obtain Sedum sarmentosum aqueous extract; (2) The aqueous extract of Sedum sarmentosum is sequentially extracted at room temperature using a first solvent and a second solvent to obtain Sedum sarmentosum extract, wherein the extraction ratio is 1:(1-10), preferably 1:(1-9), even more preferably 1:(1-8), further preferably 1:(1-7), more preferably 1:(1-6), for example 1:1, 1:1.5, 1:2, 1:2.5, 1:3, 1:3.5, 1:4, 1:4.5, 1:5, 1:5.5, 1:6, wherein the first solvent is preferably petroleum ether, and the second solvent is preferably chloroform; (3) The extract of Sedum sarmentosum was purified by column chromatography and separated by high performance liquid chromatography to obtain dehydroepiandrosterol.

[0041] It is understood that the extraction, separation, and purification in the preparation method of the present invention can be performed once or several times, such as 2, 3, 4, or 5 times.

[0042] The present invention also provides a method for increasing the content of α-SMA or CD31 in cells in vitro, which includes the step of contacting cells with dehydroepiandrosterone, wherein the cells are not particularly limited, and examples include but are not limited to RAW264.7 cells, HUVEC cells, HaCAT cells, etc.

[0043] In a preferred embodiment, the method for increasing the content of α-SMA or CD31 in cells in vitro includes culturing cells in a culture medium for 2-6 generations and counting (0.5-5)*102. 6Cells were plated and inoculated with 0.1-5 mg / ml (preferably 0.1-4.5 mg / ml, even more preferably 0.1-4 mg / ml, further preferably 0.1-3.5 mg / ml, more preferably 0.1-3 mg / ml, for example 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.2, 1.4, 1.6, 1.8, 2, 2.2, 2.4, 2.6, 2.8, 3, 3.2, 3.5 mg / ml) of dehydroepiandrosterol and cultured for 24-100 hours. The content of α-SMA or CD31 was then detected.

[0044] Example I. Establishment of Animal Models 1. Diabetic mouse model Eight-week-old C57 mice were fed a high-fat diet for one month. They were fasted for 12 hours before modeling, but water was allowed. Streptozotocin 100 mg / kg was injected intraperitoneally for two consecutive days, followed by a second injection of 50 mg / kg for one day. Blood glucose levels were monitored for two weeks after the end of the treatment. The exposed tail vein was disinfected with 75% alcohol, and the vein was cut open with sterile scissors. The first drop of blood was wiped away with a sterile cotton ball, and the second drop of blood was randomly used for blood glucose testing. Successful modeling was defined as three consecutive fasting blood glucose tests ≥11.1 mmol / L.

[0045] 2. Diabetic wound mouse model All mice were anesthetized by inhalation of 1% isoflurane. Complete anesthesia was indicated when the mice's muscles relaxed, they rolled onto their sides, and did not spontaneously return to a supine position. Skin wound models were then prepared. The specific steps are as follows: (1) Skin preparation and hair removal: Fix the mouse on a sterile operating table and remove hair in the spinal region on the back of the mouse to fully expose the skin of the surgical area.

[0046] (2) Marking: Mark the area to be excised along the midline of the spine on the back with a drawing ruler and marker, with a diameter of 8 mm.

[0047] (3) Disinfection: Disinfect the skin of the surgical area on the back three times with sterile gauze containing 75% ethanol.

[0048] (4) Wound preparation: Under sterile conditions, ophthalmic scissors and surgical forceps are used to cut away the skin in the marked area to create a circular wound with a diameter of 8 mm.

[0049] (5) Hemostasis and bandaging: Use sterile cotton swabs to stop the bleeding from the wound, clean the wound with saline, seal the wound with Vaseline gauze and medical gauze, and fix the outer layer with adhesive tape to prevent it from falling off. This is the model of a full-thickness skin defect wound.

[0050] (6) Awakening: Replace the bedding and drinking water in the mouse cage, place the mouse on a small electric blanket to warm it, accelerate the metabolism of the anesthetic drug, and promote the mouse's awakening. After awakening, place the mouse in a cage with clean bedding. Once all the mice have awakened, return them to the designated cage rack.

[0051] II. Drug Extraction and Screening 1. Drug extraction Sedum sarmentosum is a plant belonging to the Crassulaceae family. Sedum sarmentosum The fresh or dried whole herb of *Sedum sarmentosum*. Take *Sedum sarmentosum* and extract with water at a ratio of 1:10, decocting twice for 60 minutes each time to obtain an aqueous extract. Separately, take *Sedum sarmentosum* and extract with 60% ethanol at a ratio of 1:10, decocting twice for 60 minutes each time to obtain a 60% ethanol extract. Then, take another *Sedum sarmentosum* and extract with 100% ethanol at a ratio of 1:10, decocting twice for 60 minutes each time to obtain a 100% ethanol extract.

[0052] 2. Animal experiments The solvents of the three types of extracts were evaporated and concentrated to 1 g / ml. 100 μl was administered to each mouse via an 8 mm diameter wound daily for 12 consecutive days. The model group consisted of diabetic mice, and the control group consisted of normal mice. Both groups received 100 μl of physiological saline in the wound daily. Results are as follows: Figure 2 As shown, the water extract yields the best results.

[0053] The most effective aqueous extract was sequentially extracted with petroleum ether, chloroform, ethyl acetate, and water-saturated n-butanol at a 1:3 ratio until the extract was colorless, yielding five distinct layers (petroleum ether, chloroform, ethyl acetate, n-butanol, and aqueous layer). Organic reagents were removed by rotary evaporation, and the extract was reconstituted with water. The drug was prepared at a concentration of 1 g / ml, and β-FGF at 5 μg / ml. 100 μl of each drug was administered to each mouse daily via an 8 mm diameter wound for 8 consecutive days. The positive control group was prepared at 5 μg / ml. Qualitative and quantitative results are shown below. Figure 3 and Figure 4 As shown, the chloroform layer produces the best results.

[0054] 3. Cell experiments RAW264.7 cells, HUVEC cells, and HaCAT cells were cultured at high glucose levels (33 mmol / L) for at least three consecutive passages. RAW264.7 cells were counted at a rate of 1*10-1. 6 Five different layers of extract (based on a uniform amount of crude drug) were added to each cell plate. After culturing for 48 hours, the cell supernatant was collected, centrifuged at 3000 rpm, and stored at -80℃.

[0055] The above-mentioned centrifuged solution was added at a dose of 10 μl to HUVEC and HaCAT cells. Cell viability was detected using a CCK-8 assay, and cell migration rate was detected using a cell scratch assay. The results are as follows: Figure 5 As shown, the results of the cell experiments were consistent with those of the animal experiments.

[0056] 4. Screening of active ingredients 4.1 Silica gel column chromatography 200-mesh silica gel was activated in a 110°C oven for 2 hours, cooled, and then sealed for later use. A glass chromatography column (inner diameter 3 cm, length ≥ 30 cm) was prepared. The silica gel was wet-packed by mixing the activated silica gel with petroleum ether (eluent) at a 1:1 ratio and stirring to form a bubble-free homogenate. The homogenate was slowly poured into the column along a glass rod. The column walls were gently tapped with a rubber rod or a vacuum pump was connected to ensure a tight, unbroken silica gel layer (final height approximately 15 cm). A 1 cm layer of quartz sand was placed on the silica gel surface to prevent sample loading from disturbing the silica gel layer.

[0057] Dissolve the sample in ethyl acetate and slowly add it along the tube wall into the silica gel column. Use ethyl acetate and petroleum ether as eluent (volume ratio of 1:1) to elute. Collect one tube every 5 ml and collect the first 100 tubes.

[0058] Identification was performed using thin-layer chromatography with silica gel F. 254 Plates were activated in a 110℃ oven for 2 hours, cooled, and then sealed for later use. An ethyl acetate to petroleum ether ratio of 1:1 was used for development. The plates were separated into six different fractions according to polarity: tubes 1-30, 31-43, 44-60, 61-70, 71-75, and 76-100.

[0059] 4.2 Cell screening experiment Each of the above tubes was used for cell experiments with a uniform amount of crude drug. The results are as follows: Figure 6 As shown, the components in tubes 31-43 have the best effect.

[0060] 4.3 Purification The components in tubes 31-43 were further separated using an Agilent A1200 liquid chromatograph. The mobile phase was: A: water; mobile phase B: methanol; column: Agilent C18 Plus 4.6*250 mm; flow rate: 1 ml / min; B: 5%-100%, 0-50 min.

[0061] The peak with the highest concentration was extracted and purified using preparative liquid chromatography.

[0062] 4.4 Effect Verification The effects of the above-mentioned purified product were verified using cell experiments, and the results are as follows: Figure 9 As shown.

[0063] 4.5 Structural Analysis The pure sample was analyzed using high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), and the calculated molecular weight was 222, with the molecular formula C22. 13 H 18 O3.

[0064] 1 H-NMR (500 MHz, CD3OD) data as follows Figure 10 Shown: δ6.99 (1H, d, J=10 Hz, H-7), 6.43 (1H, d, J=10 Hz, H-8), 5.93 (1H, m, H-4), 2.60 (1H, d, J=15 Hz, H-2a), 2.27 (1H, d, J=15 Hz, H-2b), 2.30 (3H, s, H-10), 1.89 (3H, s, H-11), 1.06 (3H, s, H-12), 1.01 (3H, s, H-13). 1 H-NMR spectra showed four groups of sp3 methyl signals (H-10, H-11, H-12, H-13), one group of sp3 methylene signals (H-2), and three groups of sp2 methine signals (H-4, H-7, H-8).

[0065] 13 C-NMR (125 MHz, CD3OD) data as follows Figure 11 As shown: δ200.69(C-3), 200.39(C-9), 164.68(C-5), 148.38(C-7), 131.76(C-8), 128.08(C-4), 80.01(C-6), 50.57(C-2), 42.69(C-1), 27.67(C-10), 24.78(C-13), 23.56(C-12), 19.20(C-11).

[0066] The above results indicate that the isolated pure product is dehydroepiandrosterol.

[0067] 4.6 Animal Experiments Animal experiments were conducted using low (0.35 mg / ml), medium (0.7 mg / ml), and high (1.4 mg / ml) doses of dehydroepiandrosterol. The positive control was achieved using topical recombinant human granulocyte-macrophage stimulating factor gel at a concentration of 20 μg / ml. 2 The result is as follows Figure 12 As shown, the medium dose resulted in the best wound closure. Especially in the first 5 days, the effect was good, indicating that the drug works by targeting the inflammatory phase of post-injury healing. Wounds treated with the medium dose showed the most significant closure effect on day 14. Immunohistochemical results, etc., are as follows... Figure 13As shown, the medium-dose treatment of well-developed collagen fibers showed the most significant increase in regenerated epidermal length on day 14. In addition, the wounds in the medium-dose treatment group showed enhanced vascularization, such as larger areas of α-smooth muscle actin (α-SMA) vessels, higher density of CD31, and higher capillary density.

[0068] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. These modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Application of dehydroepiandrosterol in the preparation of drugs that improve or promote wound healing.

2. The use of dehydroepiandrosterol according to claim 1 in the preparation of a medicament for improving or promoting wound healing, characterized in that, The improvement or promotion of wound healing includes at least one of the following: (1) Increase the length of the regenerated epidermis; (2) Increase collagen surface area; (3) Promotes wound vascularization; (4) Increase capillary density.

3. The use of dehydroepiandrosterol according to claim 1 in the preparation of a medicament for improving or promoting wound healing, characterized in that, The wounds include skin wounds.

4. The use of dehydroepiandrosterol according to claim 1 in the preparation of a medicament for improving or promoting wound healing, characterized in that, The wounds include acute wounds and / or chronic wounds.

5. The use of dehydroepiandrosterol according to claim 4 in the preparation of a medicament for improving or promoting wound healing, characterized in that, The acute wounds include at least one of abrasions, lacerations, cuts, punctures, and burns, and the chronic wounds include diabetic wounds.

6. A method for preparing dehydroepiandrosterol, characterized in that, The preparation method includes the following steps: (1) Extract Sedum sarmentosum with water to obtain Sedum sarmentosum water extract; (2) Extract the aqueous extract of Sedum sarmentosum using an organic solvent to obtain Sedum sarmentosum extract; (3) The extract of Sedum sarmentosum is purified and separated to obtain the dehydroepiandrosterol.

7. The method for preparing dehydroepiandrosterol according to claim 6, characterized in that, In step (1), the material-to-liquid ratio is 1:(5-50).

8. The method for preparing dehydroepiandrosterone according to claim 6, characterized in that, In step (1), the water extraction time is 0.5-5 h.

9. The method for preparing dehydroepiandrosterol according to claim 6, characterized in that, The organic solvent includes at least one of petroleum ether, chloroform, ethyl acetate, n-butanol, ethanol, acetone, and cyclohexane.

10. A method for increasing the content of α-SMA and / or CD31 in cells in vitro, characterized in that, This includes the step of contacting cells with dehydroepiandrosterone.

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