CN115590896A

CN115590896A - Application of low molecular dextran taurine and chlorella vulgaris in combined preparation of medicine for treating skin related diseases

Info

Publication number: CN115590896A
Application number: CN202211159380.4A
Authority: CN
Inventors: 吕慧侠; 武航毅; 张振海
Original assignee: China Pharmaceutical University
Current assignee: China Pharmaceutical University
Priority date: 2022-09-22
Filing date: 2022-09-22
Publication date: 2023-01-13
Anticipated expiration: 2042-09-22
Also published as: CN115590896B

Abstract

The invention relates to a method for treating skin-related diseases by jointly preparing low-molecular dextran taurine and chlorella vulgarisThe application of the medicine is provided. Skin-related diseases involved therein include: burns and scalds, pressure sores, diabetic ulcers, atopic dermatitis; wherein, the wound or skin is moistened by adopting a low molecular dextran taurine solution, and then the chlorella is applied; wherein the low molecular dextran comprises 20, 40, 60 and the like of dextran, and the dosage of taurine is 0.01-5 percent; wherein the chlorella is applied in the form of cells, preparations and the like, and specifically comprises solution, hydrogel, patch, dressing and the like which take the chlorella as an active component; the Chlorella is applied at a concentration of 1 × 10 ³ ~1×10 ¹² cell/mL。

Description

Application of low molecular dextran taurine and chlorella vulgaris in combined preparation of medicine for treating skin related diseases

Technical Field

The invention relates to application of chlorella, in particular to application of low-molecular dextran taurine and chlorella in combined preparation of a medicine for treating skin-related diseases, and specifically relates to burn and scald, pressure sore, diabetic ulcer and atopic dermatitis.

Background

Chlorella (Chlorella vulgaris)Chlorellasp.) is a unicellular eukaryotic algae (2 to 10 μm) with extremely fast growth rate, which can resist various inhibitory growth conditions and is one of the most widely cultivated microalgae species. The chlorella contains rich nutrients such as protein, lipid, polysaccharide, amino acid, vitamin and the like, and the protein content of the chlorella reaches 50 percent, so the chlorella is developed into a human body nutritional supplement at present. In addition, the inclusion of the composition also contains growth factors and immunomodulators, and can be used as health products and functional foods for preventing diseases. Chlorella can continuously generate oxygen through photosynthesis under light conditions, and is known as a source of oxygen in the air. In addition, chlorella has antiinflammatory and antioxidant effects. The inside of the capsule contains rich sulfated polysaccharide, and the capsule can play an anti-inflammatory role by activating and regulating macrophages. In addition to sulfated polysaccharides, the pigment-protein complex in chlorella extract also has anti-inflammatory effect. At present, the inclusions identified as having the functions of resisting oxidation and scavenging free radicals include: lutein,𝛼-carotene, carotene,𝛽Carotene, ascorbic acid and𝛼-tocopherols and the like. In addition, the biologically active compounds thereof𝛽1, 3-Glucan as an active immunostimulant can reduce cholesterol in blood and has also been shown to have a preventive function against atherosclerosis and hypercholesterolemia.

Chlorella has the ability to improve the microenvironment of poor tissues of skin-related diseases, but chlorella is a living cell and has high proliferation and metabolic activity, which may further consume the nutrition of the lesion, reduce the supply of nutrition to tissue cells of the lesion, and thus hinder the treatment of diseases. The low molecular dextran is a nutritional blood volume expander and is suitable for treating hypovolemia and poor microcirculation accompanied with protein deficiency. Wherein the low molecular dextran can increase plasma colloid osmotic pressure, expand blood volume, expand capillary vessel, thereby improving blood perfusion and microcirculation of wound tissue, and the improvement of blood flow and increase of blood volume can accelerate metabolism and discharge of harmful products thereof.

Taurine is a conditionally essential amino acid of human body, and has been widely used in the fields of medicine, food additives, and the like. Experiments at present prove that 0.01% -1% of taurine can promote the proliferation of fibroblasts. In addition, taurine also has anti-inflammatory and antioxidant effects, and can reduce the formation of lipid peroxide, carbonyl protein and active oxygen by improving the activities of antioxidant enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and the like.

Therefore, a treatment method using the combination of low-molecular-weight dextran taurine and chlorella is proposed to hopefully obtain better treatment effect.

Disclosure of Invention

The invention relates to application of low-molecular dextran taurine and chlorella in preparation of a medicine for promoting wound healing.

Application of low molecular dextran taurine and chlorella in preparation of medicine for treating skin diseases is provided, which is a method for treating skin related diseases by combination of low molecular dextran taurine and chlorella, wherein the skin related diseases comprise burn, scald, pressure sore, diabetic ulcer and atopic dermatitis.

The combined treatment method comprises the steps of firstly applying low-molecular dextran taurine and then applying chlorella.

The low-molecular dextran taurine solution is characterized in that: every 1000 mL contains 20, 40 or 60 g of low molecular dextran, the auxiliary material is sodium bisulfite, and the content of taurine is 0.01% -5% w/v.

The chlorella preparation comprises a solution, hydrogel, a patch, a dressing, freeze-dried powder and the like which take chlorella as an active component.

The biomass screening range of the chlorella is 1 multiplied by 10 ³ ~ 1 × 10 ¹² cell/mL。

The preparation method of the low-molecular dextran taurine solution is characterized by comprising the following steps: dissolving the low molecular dextran and taurine in normal saline, and sterilizing.

The chlorella solution comprises but is not limited to chlorella BG-11 solution, and the preparation method is characterized in that: and centrifuging the sterile cultured chlorella with determined biomass, discarding the supernatant, washing the BG-11 culture medium for 2 times, and then re-suspending the chlorella by using a quantitative BG-11 culture medium to obtain a chlorella BG-11 solution.

The chlorella hydrogel comprises but is not limited to chlorella hyaluronic acid hydrogel, and the preparation method is characterized in that: mixing 10 ml of 1X 10 ⁸ The aseptically cultured chlorella was centrifuged at cell/ml concentration, the supernatant was discarded, washed 2 times with BG-11 medium and resuspended with a fixed amount of BG-11 medium. Then, 0.2g of hyaluronic acid powder was added to the chlorella solution, and stirred using a magnetic stirrer under room temperature conditions to form a chlorella hyaluronic acid hydrogel.

The low-molecular-weight dextran taurine and the chlorella are combined, and the combined treatment effect is superior to that of single application.

The low-molecular-weight dextran taurine and the chlorella are combined for use, and the effect of combined treatment can be obviously improved by firstly applying the low-molecular-weight dextran taurine and then applying the chlorella.

Burns and scalds generally refer to tissue damage caused by heat, mainly to the skin and/or mucous membrane, and serious people can also damage subcutaneous or/and submucosal tissues, such as muscles, bones, joints and even internal organs. After the burn and scald, the organism is seriously oxidized and damaged due to tissue ischemia reperfusion injury and a large amount of body fluid loss, so a large amount of active oxygen metabolites are generated and are accompanied with severe inflammatory reaction. In addition, burns and scalds can cause extensive physiological barrier damage to the body, a large amount of tissue necrosis and reduction of immune function, and meanwhile, a large amount of protein-rich exudate exists in the burn wound, so that the probability of the body being invaded by pathogens is improved. The low molecular dextran and taurine can provide nutrition required for regeneration of wound tissues, the chlorella can obviously reduce oxidative stress and inflammation, has the function of purifying exudate by metabolic protein, and can provide growth factors required for wound healing, so that the combination of the low molecular dextran and the taurine can be a potential treatment strategy.

Pressure sores are also called pressure ulcers and pressure sores, and are caused by the fact that local tissues are pressed for a long time to cause the defects of continuous blood circulation obstruction, oxygen deficiency and nutrient deficiency. Skin pressure sores are a common problem in the long-term treatment process of rehabilitation, nursing and the like, and the clinical treatment is difficult to a certain degree, and about 6 million people die of pressure sores every year. The low molecular dextran and the taurine can provide nutrition required by regeneration for wound tissues; chlorella can be used for promoting blood vessel regeneration, providing oxygen, providing nutrition, relieving inflammation of sore surface, and promoting healing of pressure sore.

Diabetic ulcers are chronic wounds caused by long-term hyperglycemia, with long treatment periods, high recurrence rates, and even risk of amputation and death. At present, the number of patients in the whole world is as high as 2600 million, which causes great medical burden. Research shows that 11.2 percent (about 1.298 hundred million) of diabetic patients in adults in China account for the incidence of diabetic ulcers of about 4.1 percent. Diabetic chronic wounds are often characterized by high glucose, high oxidative stress, severe hypoxia, high lactic acid content, and malnutrition. The low molecular dextran and the taurine can provide nutrition required by regeneration for wound tissues and can improve insulin resistance, and the combination of the low molecular dextran and the taurine is combined with the characteristics of the chlorella, so that the combination of the low molecular dextran and the taurine seems to be a feasible method for improving the microenvironment of diabetic ulcer tissues.

Atopic dermatitis is a chronic, recurrent inflammatory skin disease. Global disease burden studies have shown that the prevalence of childhood is 15% to 20% and that of adults is as high as 10%, making atopic dermatitis the 15 th most common non-lethal disease and also the most economically burdened dermatological disease. It is usually manifested as itching, redness, swelling, dryness of the skin. The low molecular dextran and the taurine can provide nutrition required by metabolism for inflammation sites, and the chlorella can be used for providing oxygen and nutrient substances and a certain humid environment to accelerate the repair process of damaged skin, so that the combination of the low molecular dextran and the taurine seems to be a feasible method for treating atopic dermatitis.

Advantageous effects

The low molecular dextran is commonly used for treating diseases with reduced blood volume and poor microcirculation accompanied with protein deficiency, has limited effect although having certain therapeutic effect on wound healing, and does not show remarkable therapeutic advantages in the treatment of skin-related diseases. Taurine has anti-inflammatory and antioxidant effects, and can reduce insulin resistance of diabetic tissues. Chlorella has the ability to improve the microenvironment of tissues with skin-related disorders, but is also subject to certain difficulties.

The key points of the invention are as follows: the low molecular dextran taurine and the chlorella vulgaris are combined for use, so that the burn, scald, pressure sore, diabetic ulcer and atopic dermatitis can be effectively treated, the treatment effect of the combined application is obviously higher than that of the single application, a synergistic interaction effect is generated, and particularly, the low molecular dextran taurine is applied firstly and then the chlorella vulgaris is applied, so that the treatment effect is more obvious.

1. The low-molecular-weight dextran taurine is combined with the chlorella, the effective components are cheap and easy to obtain, the preparation process is simple, and the method has industrial production potential.

2. The low molecular dextran taurine is combined with chlorella to achieve remarkable treatment effect, and can effectively treat burns, scalds, pressure sores, diabetic ulcers and atopic dermatitis.

3. The low molecular dextran taurine is combined with the chlorella, and the treatment effect is obviously higher than that of the low molecular dextran taurine or the chlorella which is independently applied.

4. In the combined application, the low-molecular dextran taurine is applied first and then the chlorella is applied, so that the combined application has more remarkable treatment effect.

Drawings

FIG. 1 shows that in examples 1 and 2, low-molecular dextran taurine and chlorella are combined and sequentially applied to the application of the low-molecular dextran taurine and the chlorella in the treatment of burns and scalds;

FIG. 2 shows the combined application of low molecular weight dextran taurine and chlorella in example 1 and example 2, and the application of low molecular weight dextran taurine and chlorella in pressure sore treatment in sequence;

FIG. 3 shows the combined application of low molecular weight dextran taurine and chlorella in example 1 and example 2, and the application of low molecular weight dextran taurine and chlorella in the treatment of diabetic ulcer;

FIG. 4 shows the combined use of low molecular weight dextran taurine and chlorella vulgaris in example 1 and example 2, and the use of low molecular weight dextran taurine and chlorella vulgaris in the treatment of atopic dermatitis in sequence.

Detailed Description

The present invention is further illustrated by the following specific examples, but the present invention is not limited to the contents contained in the following examples.

Example 1: preparation of low-molecular dextran taurine solution

Taking a certain amount of low molecular dextran and taurine, adding into normal saline (each 1000 mL of normal saline contains 60.0 g of dextran 40 and 1 g of taurine), stirring, and sterilizing to obtain the low molecular dextran taurine solution.

Example 2: preparation of Chlorella hydrogel

Taking 10 ml of 1X 10 ⁸ Aseptically cultured chlorella of cell/ml concentration, centrifuged (4000 rpm,3 min), the supernatant discarded, washed 2 times with BG-11 medium and resuspended in 10 ml BG-11 medium. Then, 0.2g of HA powder was added to the chlorella solution, and stirred using a magnetic stirrer (500 rpm,4 h) under room temperature conditions to form a chlorella hyaluronic acid hydrogel.

Example 3: in vivo evaluation of treatment of burns and scalds

Selecting 6-8 week old Balb/C male mice, and using a constant temperature and voltage controlled electric iron instrument to make a mouse scald model. After molding, mice were randomly divided into five groups: burn and scald control groups (wound is sealed by 3M dressing), example 1 groups (80 muL of low-molecular dextran taurine solution is added, and wound is sealed by 3M dressing), example 2 groups (80 muL of chlorella hydrogel is added, and wound is sealed by 3M dressing), combined application groups (40 muL of low-molecular dextran taurine solution is added, 40 muL of chlorella hydrogel is added at the same time, and wound is sealed by 3M dressing), and sequential application groups (40 muL of low-molecular dextran taurine solution is added firstly, 40 muL of chlorella hydrogel is added after 1 h, and wound is sealed by 3M dressing). The hydrogels were changed daily for all experimental groups, and the wounds were photographed on

days

0, 3, 7, and 10 and quantitatively analyzed by ImageJ software.

The result is shown in figure 1, the combined application of the low molecular dextran taurine and the chlorella can effectively treat the burns and scalds, and the combined treatment effect is obviously higher than that of the single application. Because the administration volume of the combined application group is unchanged (the total volume of the low-molecular-weight dextran taurine and the chlorella is the same as that of the group in example 1 and the group in example 2), but a better treatment effect is obtained, the combination of the low-molecular-weight dextran taurine and the chlorella is considered to have a certain synergistic effect in treating the burns and scalds. In addition, the application of the low-molecular-weight dextran taurine and then the chlorella hydrogel has better treatment effect.

Example 4: in vivo evaluation of treatment of pressure sores

200 g of SD male rats are selected, and a rat pressure sore model is manufactured by a mechanical compression method. After molding, mice were randomly divided into five groups: a pressure sore control group (sealing a wound by adopting a 3M dressing), an example 1 group (adding 80 mu L of low-molecular dextran taurine solution and sealing the wound by adopting the 3M dressing), an example 2 group (adding 80 mu L of chlorella hydrogel and sealing the wound by adopting the 3M dressing), a combined application group (adding 40 mu L of low-molecular dextran taurine solution, simultaneously adding 40 mu L of chlorella hydrogel and sealing the wound by adopting the 3M dressing), and a sequential application group (firstly adding 40 mu L of low-molecular dextran taurine solution, adding 40 mu L of chlorella hydrogel after 1 h and sealing the wound by adopting the 3M dressing). The hydrogels were changed daily for all experimental groups, and the wounds were photographed on

days

0, 3, 7, 10 and quantitatively analyzed by ImageJ software.

The result is shown in fig. 2, the low molecular dextran taurine and the chlorella vulgaris can be used for effectively treating the pressure sore by combined application, and the combined treatment effect is obviously higher than that of the single application. Because the administration volume of the combined application group is unchanged (the total volume of the low-molecular-weight dextran taurine and the chlorella is the same as that of the group in example 1 and the group in example 2), but a better treatment effect is obtained, the combination of the low-molecular-weight dextran taurine and the chlorella for treating the pressure sore has certain synergistic effect. In addition, the application of the low-molecular-weight dextran taurine and then the chlorella hydrogel has better treatment effect.

Example 5: in vivo evaluation of treatment of diabetic ulcers

After 4 weeks of feeding of 4-6 weeks old Balb/C male diabetic mice with the diet (high fat and high sugar) for 4 weeks, streptozotocin citrate buffer solution (streptozotocin: 55 mg/kg) was intraperitoneally injected for 5 consecutive days. Mice with post-detection blood glucose of 16.7 mM or more were considered successful in modeling. Mice were anesthetized with an intraperitoneal injection of chloral hydrate and the dorsal hairs were shaved off. A round full-thickness skin wound was made using a biopsy punch with an 8 mm inner diameter, and a silicone pad was sutured around the wound tissue using non-absorbable sutures to prevent contraction of the mouse skin, hydrogel was added, and a clear polyurethane film was selected as the outer lining to form a closed system. Mice were randomized into five groups in the experiment: a diabetes control group (sealing a wound by adopting a 3M dressing), an example 1 group (adding 80 mu L of low-molecular-weight dextran taurine solution and sealing the wound by adopting the 3M dressing), an example 2 group (adding 80 mu L of chlorella hydrogel and sealing the wound by adopting the 3M dressing), a combined application group (adding 40 mu L of low-molecular-weight dextran taurine solution, simultaneously adding 40 mu L of chlorella hydrogel and sealing the wound by adopting the 3M dressing), and a sequential application group (firstly adding 40 mu L of low-molecular-weight dextran taurine solution, adding 40 mu L of chlorella hydrogel after 1 h and sealing the wound by adopting the 3M dressing). All experimental groups were changed daily with hydrogel. Wounds were photographed on

days

0, 3, 7, 9 and quantitatively analyzed by ImageJ software.

The result is shown in figure 3, the combined application of the low molecular dextran taurine and the chlorella can effectively treat the diabetic ulcer, and the combined treatment effect is obviously higher than that of the single application. Because the administration volume of the combined application group is unchanged (the total volume of the low-molecular-weight dextran taurine and the chlorella is the same as that of the group in example 1 and the group in example 2), but a better treatment effect is obtained, the combination of the low-molecular-weight dextran taurine and the chlorella for treating the diabetic ulcer is considered to have a certain synergistic effect. In addition, the chlorella hydrogel is applied after the low-molecular dextran taurine is applied, so that the treatment effect is better.

Example 6: in vivo evaluation for the treatment of atopic dermatitis

A base solution was prepared using acetone and olive oil at a ratio of 3. After molding, mice were randomly divided into five groups: an atopic dermatitis control group (a wound is sealed by adopting a 3M dressing), an example 1 group (80 mu L of low-molecular dextran taurine solution is added, and the wound is sealed by adopting the 3M dressing), an example 2 group (80 mu L of chlorella hydrogel is added, and the wound is sealed by adopting the 3M dressing), a combined application group (40 mu L of low-molecular dextran taurine solution is added, 40 mu L of chlorella hydrogel is simultaneously added, and the wound is sealed by adopting the 3M dressing), and a successive application group (40 mu L of low-molecular dextran taurine solution is firstly added, 40 mu L of chlorella hydrogel is added after 1 h, and the wound is sealed by adopting the 3M dressing). Chlorella hydrogel is then used for treatment for a week, three times a day, for a total of four weeks. After the experiment was completed, the mouse was sacrificed by removing the neck, and the ear and back skin were immersed in 4% paraformaldehyde fixing solution for pathological sectioning.

Dermatitis scores were performed on the skin lesion areas on the backs of the mice at predetermined time points (0, 7, 14, 21, 25, 28). The specific criteria are as follows: no symptoms, score 0; mild symptoms, score 1; moderate symptoms, score 2; severe symptoms, score 3. Erythema/bleeding, edema, scratch/erosion and scaling/dryness symptoms were evaluated separately, and the dermatitis score was calculated as the sum of the individual scores, with a full score of 12.

The results are shown in fig. 4, the combined application of low molecular dextran taurine and chlorella vulgaris can effectively treat atopic dermatitis, and the combined treatment effect is remarkably higher than that of the single application. Because the administration volume of the combined application group is unchanged (the total volume of the low-molecular-weight dextran taurine and the chlorella is the same as that of the group in example 1 and the group in example 2), but better treatment effect is obtained, we think that the low-molecular-weight dextran taurine and the chlorella are combined to treat the atopic dermatitis to have a certain synergistic effect. In addition, the application of the low-molecular-weight dextran taurine and then the chlorella hydrogel has better treatment effect.

Claims

1. The application of the low-molecular-weight dextran taurine and chlorella in preparing the medicine for promoting wound healing is disclosed.

2. The low molecular dextran taurine and chlorella are combined to be used for preparing the medicine for treating skin diseases.

3. The use according to claim 2, wherein said skin diseases include burns, scalds, pressure sores, diabetic ulcers, atopic dermatitis.

4. The use according to any one of claims 1-3, wherein the low molecular dextran taurine is applied first, followed by the chlorella.

5. The use as claimed in claim 4, wherein the chlorella is formed into a dressing.

6. The use according to claim 5, wherein the dressing is a cell solution, hydrogel or patch.

7. The use according to claim 6, wherein the hydrogel is prepared from Chlorella vulgaris and hyaluronic acid.

8. The use of claims 1-7, wherein the low molecular dextran taurine solution contains 60.0 g of low molecular dextran 20, 40 or 60 per 1000 mL, the auxiliary material is sodium bisulfite, the taurine content is 0.01% -5% w/v, the low molecular dextran and taurine are dissolved in normal saline, and the sterilization is performed.

9. The use of claim 7, wherein the chlorella biomass is selected from the range of 1 x 10 ³ ~ 1×10 ¹² cell/mL。

10. The use according to claim 9, wherein the chlorella hydrogel comprises but is not limited to chlorella hyaluronic acid hydrogel, and the preparation method is characterized in that: mixing 10 ml of 1X 10 ⁸ centrifuging sterile cultured chlorella with cell/ml concentration, discarding supernatant, washing BG-11 culture medium for 2 times, and re-suspending with quantitative BG-11 culture medium;

then, 0.2g of hyaluronic acid powder was added to the chlorella solution, and stirred using a magnetic stirrer under room temperature conditions to form a chlorella hyaluronic acid hydrogel.