Oyster shell hydrogen powder composition with pH value buffering function and preparation method thereof

Abstract

The invention discloses an oyster shell hydrogen powder composition with a pH value buffering function, which is prepared from the following components in percentage by mass: 40 to 50 percent of oyster shell hydrogen powder, 20 to 25 percent of starch, 10 to 15 percent of citric acid and 15 to 20 percent of hydroxyethyl methyl cellulose, and the sum of the percentage of the components is 100 percent. The invention also discloses a preparation method of the oyster shell hydrogen powder composition. According to the composition and the preparation method disclosed by the invention, the problem that Ca (OH) 2 generated when an existing product containing oyster shell hydrogen powder releases H2 is strongly alkaline and can generate obvious stimulation and damage to gastrointestinal mucosa is solved.

Description

Technical Field

[0001] The invention belongs to the technical field of medicines and health products, and in particular relates to an oyster shell hydrogen powder composition with a pH buffering function. The invention also relates to a preparation method of the composition. Background Art

[0002] Oyster shell powder (OSP), also known as shell powder, is made from dried and ground mussel shells, oyster shells, clam shells, and snail shells. Its main ingredients are calcium carbonate, 96.4% calcium gluconate, 38.0% pure calcium, and trace elements such as magnesium, iron, manganese, and phosphorus. Main effects: ① Calcium supplementation: OSP has a high calcium content, which can promote bone development, improve bone toughness and density, and effectively prevent osteoporosis; ② Pain relief: OSP is a traditional Chinese medicine that can astringe and relieve pain. It can effectively inhibit the secretion of free acid, reduce the acidic stimulation of the gastric mucosa, prevent gastroenteritis, and accelerate the healing of ulcers; ③ Sedation: The active ingredients in OSP can directly act on the central nervous system to produce a sedative and tranquilizing effect, effectively improving symptoms such as insomnia and neurasthenia.

[0003] H2 is the smallest molecule in nature. It is colorless, odorless and harmless. It has strong penetrating power, rapid diffusion and other characteristics. It can penetrate various biological barriers and has extremely high biosafety. In recent years, H2 has attracted more and more attention as a molecule with a wide range of biological activities. From more than 1,000 hydrogen medical research papers at home and abroad, we can see that H2 has many health care effects on the human body. The most basic effect is selective antioxidant effect, which helps to slow down the aging process of the skin and has a beauty and anti-aging effect. It helps to regulate metabolism and relieve metabolic diseases. It helps to activate the immune system to enhance resistance, helps to inhibit cell mutation and reduce cell apoptosis, helps to promote the repair and regeneration of damaged tissues, helps to reduce the degree of inflammatory response, helps to stabilize cell membranes, reduces allergic symptoms and reduces radiation damage, helps to relieve fatigue, and improves work efficiency and quality of life. In the "National Food Safety Standard for the Use of Food Additives" (GB2760-2014) issued in 2014, H2 is listed as a food additive. It can enter the body through inhalation, oral administration, etc., and freely bind to cells in the body to exert its biological properties of antioxidant, anti-inflammatory and anti-apoptotic. With the development of hydrogen molecular medicine, H2 as a medical gas has received extensive attention in the therapeutic research of various diseases. The amount of H2 produced by the human body is limited. Under normal circumstances, the human body can produce about 150 ml of H2 per day, which is mainly produced by intestinal hydrogen-producing bacteria fermenting unabsorbed carbohydrates, which is difficult to meet long-term antioxidant needs. Although H2 has broad application prospects in the medical field, the existing preparation methods of hydrogen-rich water, hydrogen-rich beverages and other products mainly produce H2 by electrolyzing water or by reacting metal magnesium with water, etc., all of which have problems such as unstable amount of dissolved H2 and easy escape.

[0004] Oyster shell hydrogen powder (OSHP) is a product that is enriched with a certain amount of hydrogen (H2) through a special physical process. The main component of H2 is calcium hydride (Ca H2), also known as hydrogenated OSP. In addition to retaining the calcium-replenishing and bone-strengthening, astringent and analgesic, and sedative effects of OSP, OSHP can release more H2 after oral administration and contact with water, increasing the antioxidant, anti-inflammatory, immunity-enhancing, blood circulation-promoting, sleep quality-improving, wound healing and other health-care effects of H2. Studies have shown that OSHP can continuously release H2 when it is in contact with water in vitro or in vivo, and both the amount of H2 released and the duration of continuous H2 release are higher than various hydrogen-rich water (H2-rich aqueous solutions) sold on the market. Unfortunately, OSHP reacts chemically with water (H2O) according to the chemical equation: CaH2+2H2O=2H2↑+Ca(OH)2. The Ca(OH)2 produced while releasing H2 is strongly alkaline, which can significantly irritate and damage the gastrointestinal mucosa. Its safety limits the oral development and application of OSHP. Summary of the invention

[0005] The first object of the present invention is to provide an oyster shell hydrogen powder composition with a pH buffering function, which solves the problem that the existing oyster shell hydrogen powder products release H2 and produce Ca(OH)2 which is strongly alkaline and causes obvious irritation and damage to the gastrointestinal mucosa.

[0006] The second object of the present invention is to provide a method for preparing the above composition.

[0007] The first technical solution adopted by the present invention is: an oyster shell hydrogen powder composition with a pH buffering function is composed of the following components by mass percentage: 40% to 50% oyster shell hydrogen powder, 20% to 30% starch, 10% to 15% citric acid, and 15% to 20% hydroxyethyl methyl cellulose, and the sum of the percentages of the above components is 100%

[0008] The first technical solution adopted by the present invention is also characterized in that:

[0009] Furthermore, the oyster shell hydrogen powder composition with the function of buffering pH is composed of the following components by mass percentage: 45% oyster shell hydrogen powder, 24% starch, 13% citric acid, and 18% hydroxyethyl methylcellulose.

[0010] Furthermore, citric acid is used as an acidity regulator to adjust the pH value of the composition to a stable neutral state.

[0011] Furthermore, hydroxyethyl methylcellulose is used as a sustained-release preparation to improve the stability of the composition and enhance the safety of the composition.

[0012] Furthermore, the oyster shell hydrogen powder composition is packaged in capsule shells, the capsule shells are made of edible and easily digestible gelatin, and are made into capsules with a content of 0.45 g / capsule.

[0013] The second technical solution adopted by the present invention is: a method for preparing an oyster shell hydrogen powder composition with a pH buffering function, which is prepared according to the following steps:

[0014] Step 1, sieve oyster shell hydrogen powder, starch, citric acid and hydroxyethyl methylcellulose through a 100-mesh sieve for later use;

[0015] Step 2: According to the formula, accurately weigh oyster shell hydrogen powder, starch, citric acid, and hydroxyethyl methylcellulose and mix them;

[0016] Step 3, filling the total mixed material into hollow capsules;

[0017] Step 4: Pack the filled capsules.

[0018] The beneficial effects of the present invention are:

[0019] 1. The oyster shell hydrogen powder composition of the present invention has significant anti-inflammatory and wound healing effects, and can provide hydrogen to the body in a long-term and stable manner. By taking an OSHP prescription combination with a buffered pH value orally, the H2 content in the human body can be effectively increased, thereby playing an anti-inflammatory and wound growth and healing role. In addition, an OSHP prescription combination with a buffered pH value also has the advantages of high safety and few side effects, and is suitable for long-term use by the general population.

[0020] 2. The present invention utilizes solid-state hydrogenation technology to produce a hydrogen-rich OSHP, which has the characteristic of slowly and continuously releasing hydrogen when in contact with water. After external application, it can slowly and continuously release hydrogen on the body surface to promote rapid healing of wounds.

[0021] 3. The present invention utilizes solid-state hydrogenation technology to produce a H2-rich oyster shell powder, which has the characteristic of slowly and continuously releasing H2 after contacting water. Oral administration through the gastrointestinal tract can slowly and continuously release H2 with certain health-care effects, thereby achieving significant anti-inflammatory, immune and metabolic regulation effects and promoting rapid wound healing.

[0022] 4. The composition of the present invention uses OSHP, citric acid, hydroxyethyl methylcellulose (HEMC), and starch as the contents of an OSHP prescription combination with a buffered pH value. The characteristic of OSHP in releasing H2 for a long time is utilized, and citric acid (CA) is added for acid-base buffering, so that the pH value of an OSHP prescription combination for anti-inflammatory, immune regulation, metabolism regulation, and rapid wound healing is adjusted to 7. Hydroxyethyl methylcellulose (HEMC) is then added to improve the stability of the drug and increase the sustained release effect of the drug, thereby achieving the purpose of enhancing the safety of the drug.

[0023] 5. The oyster shell hydrogen powder composition with a buffered pH value of the present invention can immediately neutralize and buffer the strong alkaline substance Ca(OH)2 originally produced after oral administration of OSHP, generating calcium citrate (Ca(C6H5O6)2) as organic calcium that is more conducive to the body's absorption, and while releasing H2, the liquid pH is always maintained at about 7, and Ca(OH)2 no longer causes irritation and damage to the gastrointestinal mucosa. The present invention proposes an OSHP prescription composition with a pH buffering function. This technical feature allows the biological health care effect of H2 to be scientifically combined with the calcium supplement effect to produce a synergistic health care effect. The technology is highly mature, and the results of small-scale tests have confirmed that it has met the requirements of large-scale industrial production.

[0024] 6. The powdering process of the preparation method of the composition of the present invention adopts a mechanical powdering method, and the raw materials such as oyster hydrogen powder and citric acid are powdered with a high-speed grinder. This method is faster and more efficient than the traditional method, and can also ensure that the powder is uniform and fine. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 is a trend diagram of H2 release concentration of different contents of OSHP in the composition of Example 1 of the present invention over 26 hours;

[0026] Figure 2 is a 26-hour pH value variation trend diagram of the composition of Example 1 of the present invention when different contents of OSHP release H2;

[0027] Figure 3A It is a comparative observation diagram of the anti-inflammatory and anti-allergic effects of OSHP and dexamethasone in the composition of the present invention on mice with atopic dermatitis;

[0028] Figure 3B is a schematic diagram of the comparison results of the effects of OSHP and dexamethasone in the composition of the present invention on the scores of atopic dermatitis mice;

[0029] Figure 3C is a schematic diagram of the comparison of the effects of OSHP in the composition of the present invention and dexamethasone on the scratching frequency of mice with atopic dermatitis;

[0030] Figure 3Dis a schematic diagram of the comparison results of the effects of OSHP in the composition of the present invention and dexamethasone on the body weight changes of mice with atopic dermatitis;

[0031] Figure 3E It is a schematic diagram of the comparison results of the effects of OSHP and dexamethasone in the composition of the present invention on the changes in spleen coefficients of mice with atopic dermatitis;

[0032] Figure 4A It is a schematic diagram of the comparison results of the effects of OSHP and dexamethasone in the composition of the present invention on the pathological morphology of skin ulcers in mice with atopic dermatitis;

[0033] Figure 4B It is a schematic diagram of the comparative results of the effects of OSHP and dexamethasone in the composition of the present invention on the degree of skin ulcers in mice with atopic dermatitis;

[0034] Figure 4C It is a schematic diagram of the comparative results of the effects of OSHP and dexamethasone in the composition of the present invention on the pathological morphology of epidermal hyperplasia in mice with atopic dermatitis;

[0035] Figure 4D is a schematic diagram of the comparative results of the effects of OSHP and dexamethasone in the composition of the present invention on the degree of epidermal hyperplasia in mice with atopic dermatitis;

[0036] Figure 5A is a schematic diagram of the comparison of the effects of OSHP and dexamethasone in the composition of the present invention on the serum IgE level of mice with atopic dermatitis;

[0037] Figure 5B is a schematic diagram of the comparison of the effects of OSHP and dexamethasone in the composition of the present invention on the serum IL-17 level in mice with atopic dermatitis;

[0038] Fig. 6A It is a comparative schematic diagram of the effects of OSHP and dexamethasone in the composition of the present invention on the absolute number of lymphocytes in the peripheral blood of mice with atopic dermatitis;

[0039] Figure 6B It is a comparative schematic diagram of the effects of OSHP and dexamethasone in the composition of the present invention on the absolute number of monocytes in the peripheral blood of mice with atopic dermatitis;

[0040] Figure 6C It is a comparative schematic diagram of the effects of OSHP in the composition of the present invention and dexamethasone on the absolute number of neutrophils in the peripheral blood of mice with atopic dermatitis;

[0041] Fig.6D It is a comparative schematic diagram of the effects of OSHP and dexamethasone in the composition of the present invention on the proportion of lymphocytes in the peripheral blood of mice with atopic dermatitis;

[0042] Fig. 6E It is a comparative schematic diagram of the effects of OSHP and dexamethasone in the composition of the present invention on the proportion of monocytes in the peripheral blood of mice with atopic dermatitis;

[0043] Fig. 6F is a comparative schematic diagram of the effects of OSHP and dexamethasone in the composition of the present invention on the proportion of neutrophils in the peripheral blood of mice with atopic dermatitis;

[0044] Figure 7 This is a schematic diagram of the observation results of the back wound recovery effects of scalded mice in each group of the present invention;

[0045] Figure 8 It is a schematic diagram of the comparison results of body weight changes of scalded mice in various groups of the present invention. DETAILED DESCRIPTION

[0046] The present invention will be further described below through the accompanying drawings and specific embodiments.

[0047] The present invention provides an oyster shell hydrogen powder composition with a buffered pH value, an OSHP buffer composition of a strong alkaline substance Ca(OH)2 in the process of CaH2+2H2O=2H2↑+Ca(OH)2 can be instantly adjusted by an acid-base buffer and a preparation method thereof, wherein the pH value can be sustainably stabilized at about 7, and the pH value is neutral. The preparation method of the present invention is simple and easy to implement, and is suitable for mass industrial production.

[0048] The present invention aims at the bottleneck problem that the existing product OSHP generates Ca(OH)2 when releasing H2, which causes obvious irritation and damage to the gastrointestinal mucosa. The present invention provides a scientific solution and preparation method verified by multiple experiments, and relates to the field of food and health products. The raw materials are: oyster shell hydrogen powder, starch, citric acid, and hydroxyethyl methylcellulose. Experiments have shown that the present invention can continuously release H2 for 26 hours after OSHP is mixed with water, and the pH value of the liquid is stable at about 7. Animal experimental studies have shown that the OSHP prescription combination significantly improves the symptoms of atopic dermatitis and has a good final therapeutic effect; the OSHP prescription combination accelerates the healing of the wounds of scalded mice, repairs the affected skin, and promotes the growth and healing of the wounds. The present invention adopts the following technical scheme: according to the specification of 0.45g / capsule of content and 1000 capsules, the components are composed of the following components by mass percentage: 40% to 50% of oyster shell hydrogen powder, 20% to 25% of starch, 10% to 15% of citric acid, and 15% to 20% of hydroxyethyl methyl cellulose, and the sum of the percentages of the above components is 100%.

[0049] The present invention also provides a method for preparing an oyster shell hydrogen powder composition with a buffered pH value, the steps of which are as follows:

[0050] (1) Pass OSHP, starch, citric acid and hydroxyethyl methylcellulose through a 100-mesh sieve respectively for later use;

[0051] (2) According to the formula, OSHP, starch, citric acid, and hydroxyethyl methylcellulose were accurately weighed and mixed;

[0052] (3) filling the total mixed material of (2) into hollow capsules;

[0053] (4) Packing the filled capsules.

[0054] The OSHP prescription combination of the present invention is a hydrogen ion-rich oyster shell extract powder produced by solid-state hydrogenation technology, which is produced domestically and can release H2 in large quantities over a long period of time after contacting water. Selection of pH regulator: In order to control the pH of the preparation and facilitate the release of H2, a pH regulator is required. Initially, sodium dihydrogen phosphate was selected, but uneven mixing occurred after addition. In addition, considering that adding a large amount of phosphorus will affect the in vivo absorption of calcium and thus affect the preparation effect, citric acid was finally selected. Citric acid is more acidic and can more effectively adjust the pH value, and the generated calcium citrate is more conducive to in vivo absorption. Under the premise of not causing stomach damage, the drug utilization rate is improved. Therefore, the innovation of the present invention is that the acidity regulator is citric acid, with the molecular formula of C6H8O6, which is an organic weak acid. When Ca(OH)2 in OSHP meets the citric acid aqueous solution, calcium citrate (Ca(C6H5O6)2) and water (H2O) produce a neutralization reaction. The chemical reaction is Ca(OH)2+2C6H8O6=Ca(C6H5O6)2+2H2O. This neutralization reaction is one-way and there is no secondary reaction. At the same time, calcium citrate is an organic calcium acid with a good absorption rate and does not cause irritation to the gastrointestinal mucosa, further improving the calcium supplementation, analgesia and anti-inflammatory effects. Hydroxyethyl methylcellulose (HEMC) is a non-ionic surfactant with auxiliary and sustained-release agent functions such as thickening, suspension, bonding, floating, dispersion, and film formation. It can play a role in improving the stability and solubility of drugs and controlling the release rate of drugs to achieve a sustained-release effect. Starch is a basic excipient for oral solid preparations. On the one hand, it acts as a filler to expand the volume, and on the other hand, it acts as a disintegrant to promote the rapid release of drugs. The bonding, dispersing and lubricating properties of starch are essential to ensure the stability and homogeneity of the preparation. Its hygroscopicity provides additional protection for the drug, effectively resisting the erosion of moisture and maintaining the stability of the drug. In addition, starch can also improve the taste of the drug. The innovation of the powdering process is the use of mechanical powdering method, using a high-speed grinder to powder the raw materials and auxiliary materials such as oyster hydrogen powder and citric acid. This method is faster and more efficient than the traditional method, while also ensuring that the powder is uniform and fine.

[0055] The technical solution of the present invention is further described below through embodiments and drawings.

[0056] Example 1

[0057] This embodiment adopts the following technical scheme: according to the specification of 0.45g / capsule of content and 1000 capsules, the components and dosages are as follows: 30 parts (45%) of oyster shell hydrogen powder, 16 parts (24%) of starch, 8.8 parts (13%) of citric acid, and 12 parts (18%) of hydroxyethyl methylcellulose.

[0058] The preparation method thereof comprises the following steps:

[0059] (1) Pass OSHP, starch, citric acid and hydroxyethyl methylcellulose through a 100-mesh sieve respectively for later use;

[0060] (2) According to the formula, OSHP, starch, citric acid, and hydroxyethyl methylcellulose were accurately weighed and mixed;

[0061] (3) filling the total mixed material of (2) into hollow capsules;

[0062] (4) Packing the filled capsules.

[0063] Example 2

[0064] This embodiment adopts the following technical scheme: according to the specification of 0.45g / capsule of content and 1000 capsules, the components and dosages are as follows: 26.7 parts (40%) of oyster shell hydrogen powder, 20 parts (30%) of starch, 10 parts (15%) of citric acid, and 10 parts (15%) of hydroxyethyl methylcellulose.

[0065] The preparation method thereof comprises the following steps:

[0066] (1) Pass OSHP, starch, citric acid and hydroxyethyl methylcellulose through a 100-mesh sieve respectively for later use;

[0067] (2) According to the formula, OSHP, starch, citric acid, and hydroxyethyl methylcellulose were accurately weighed and mixed;

[0068] (3) filling the total mixed material of (2) into hollow capsules;

[0069] (4) Packing the filled capsules.

[0070] Example 3

[0071] This embodiment adopts the following technical scheme: according to the specification of 0.45g / capsule of content and 1000 capsules, the components and dosages are as follows: 33.4 parts (50%) of oyster shell hydrogen powder, 13.4 parts (20%) of starch, 6.7 parts (10%) of citric acid, and 13.4 parts (20%) of hydroxyethyl methylcellulose.

[0072] The preparation method thereof comprises the following steps:

[0073] (1) Pass OSHP, starch, citric acid and hydroxyethyl methylcellulose through a 100-mesh sieve respectively for later use;

[0074] (2) According to the formula, OSHP, starch, citric acid, and hydroxyethyl methylcellulose were accurately weighed and mixed;

[0075] (3) filling the total mixed material of (2) into hollow capsules;

[0076] (4) Packing the filled capsules.

[0077] The effects of the composition of the present invention are verified by experiments in combination with the accompanying drawings. The following embodiments are for illustrative purposes and are not intended to limit the scope of the present invention. Other embodiments based on the embodiments of the present invention that are not innovatively developed belong to the protection scope of the present invention.

[0078] The present invention provides an OSHP prescription combination with a buffered pH value, comprising: a capsule shell made of edible and easily digestible gelatin, and a capsule with a content of 0.45 g / capsule. The components of the capsule content medicine are: oyster shell hydrogen powder, starch, citric acid, and hydroxyethyl methylcellulose.

[0079] like Figure 1 As shown, the hydrogen-enriching effect of the composition provided in Example 1 was verified:

[0080] During the experiment, the same distilled water, the same temperature, the same measuring instrument, and the same person were used to perform the same technical standard measurement to ensure that there was only one variable, which was the H2 release amount of the powder content of the OSHP prescription combination rich in hydrogen ions. Figure 1 The figure is a line graph of hydrogen release in water of a hydrogen-rich OSHP prescription combination powder at different concentrations, i.e., 1 mg / mL, 5 mg / mL, 10 mg / mL, 20 mg / mL and 100 mg / mL. It can be seen from the figure that the OSHP prescription combination can continuously release H2 in water for 26 hours. In the first 6 hours, the H2 release of different concentration groups of an OSHP prescription combination with a buffered pH value generally shows a slow upward trend. After 6 hours, the hydrogen release of different concentration groups of capsule content powder gradually decreases, and the test is 0 at the 26th hour. It can be seen from the figure that the H2 release is related to the concentration of the capsule content powder. The higher the concentration, the higher the hydrogen release. The 100 mg / mL group can reach a maximum of 523 ppb. And within 16 hours from the start of the experiment, the H2 release of the five groups of different concentration test groups can basically reach more than 200 ppb.

[0081] like Figure 2 As shown, the acid-base buffering effect of the composition provided in Example 1 was verified:

[0082] During the experiment, the same distilled water, the same temperature, the same measuring instrument, and the same person were used to measure the same technical standards to ensure that there was only one variable, pH, of the hydrogen-rich OSHP prescription combination. Figure 2 The figure is a line graph of the pH values ​​of the powder contents of a hydrogen ion-rich OSHP prescription combination in water at different concentrations, i.e., 1 mg / mL, 5 mg / mL, 10 mg / mL, 20 mg / mL and 100 mg / mL. From the figure, it can be seen that a hydrogen ion-rich OSHP prescription combination can maintain a stable pH value of 7 during the process of continuously releasing H2 in water.

[0083] Experiment 1: Experimental study on the anti-inflammatory effect of the composition of Example 1 on atopic dermatitis

[0084] Atopic dermatitis (AD) is a chronic skin disease, also known as atopic eczema or genetic allergic dermatitis. The main clinical manifestations are eczematous rashes on the local skin, accompanied by obvious itching symptoms, and fluid exudation. After scratching, there may be symptoms such as bleeding and erosion. After the fluid exudates, it will gradually form scabs and desquamation, which seriously affects the quality of life of patients and their families. At present, the treatment of AD mainly aims to avoid allergens, relieve clinical symptoms, reduce complications and recurrence rates. Mild AD patients generally use glucocorticoids (Topic corticosteroid, TCS) and calcineurin inhibitors (Topical calcineurin inhibitors, TCL), while moderate and severe patients, in addition to the maintenance use of TCS and TCL, also need to use antihistamines, cyclosporine and phototherapy to control itching symptoms. Experiment 1 established an AD mouse model by inducing 2,4-dinitrochlorobenzene (DNCB) to evaluate the improvement of the OSHP prescription combination on AD mice, providing a theoretical basis for verifying the anti-inflammatory and anti-allergic effects of the OSHP prescription combination.

[0085] 1. Reagent Preparation

[0086] Preparation of 0.5% sodium carboxymethyl cellulose: Dissolve 1 g of sodium carboxymethyl cellulose in 150 ml of hot water and place in a hot magnetic stirrer for high temperature dissolution until all solids are dissolved and then adjust the volume to 200 ml.

[0087] Acetone and olive oil mixed solution: Acetone and olive oil are evenly mixed in a ratio of 3:1 by volume.

[0088] 1% DNCB solution: Weigh 1.2 g of DNCB solid and dissolve it in a mixed solution of 12 ml of acetone and olive oil (9 ml of acetone + 3 ml of olive oil), and mix thoroughly.

[0089] 0.4% DNCB solution: Weigh 48 mg of DNCB solid and dissolve it in a mixed solution of 12 ml of acetone and olive oil (9 ml of acetone + 3 ml of olive oil), and mix thoroughly.

[0090] OSHP prescription combination solution: effective concentration 50 mg / kg / d, dissolved in 0.5% sodium carboxymethyl cellulose.

[0091] 2. DNCB animal specific dermatitis modeling, grouping and drug administration

[0092] The experimental grouping was mainly divided into 4 groups, normal group, model group, dexamethasone group, and OSHP prescription combination group. Use an electric shaver to shave 2.5cm×2.5cm of the mouse back skin, remove the mice with back skin problems to avoid interference, and use depilatory cream to remove hair twice in the shaved area to ensure that the mouse back skin can be completely exposed. After using the depilatory cream, wash with water to prevent the depilatory cream from corroding the mouse skin and affecting the modeling effect. On the first day of the experiment, the model group, the dexamethasone positive control group, and the OSHP prescription combination were smeared with 1% DNCB solution, and each was smeared with 200 microliters for sensitization. The normal group was smeared with a mixed solution of acetone and olive oil (3:1 acetone: olive oil) for 3 consecutive days. On the 8th day, 0.4% DNCB solution was applied to the back of the mouse to induce atopic dermatitis. The normal control was given a mixed solution of acetone and olive oil (3:1 acetone: olive oil) for 4 consecutive days. Drug treatment was started on the day of modeling, and each group was given drugs by gavage for 12 consecutive days, and experimental samples were collected on the 13th day.

[0093] 3. Changes in atopic dermatitis scores

[0094] According to the AD scoring standard, the severity of the skin lesions on the back of the mice was graded, and the scores were based on the presence of erythema / hemorrhage, scratches / erosion, skin edema, scaling or lichenification on the back. Each level of severity was scored from 0 to 4 points. The scores of different levels were added together to get the final score. The scores were scored every 2 days according to the above method. The higher the score, the more severe the symptoms. The average value was finally taken. The scoring content and standards are as follows:

[0095] (1) Skin erythema / hemorrhage: 4 points if the area is larger than 75% of the exposed skin area; 3 points if the area is between 40% and 75%; 2 points if the area is between 10% and 40%; 1 point if the area is less than 10%; 0 points if there is no bleeding or erythema.

[0096] (2) Skin scratches / erosions: 4 points if the area is larger than 75% of the exposed skin area; 3 points if the area is between 40% and 75%; 2 points if the area is between 10% and 40%; 1 point if the area is less than 10%; 0 points if there is no bleeding or erythema.

[0097] (3) Skin edema: dark red skin with a moist surface, obvious swelling, and no new hair was scored as 4 points; light red skin with a slightly dry surface, reduced edema, and no new hair was scored as 3 points; pink skin, slightly darker than normal mice, with an uneven surface and new hair was scored as 2 points; skin color close to the normal skin color of mice, with a large amount of new hair and smooth skin was scored as 1 point; none of the above symptoms was scored as 0 points.

[0098] (4) Skin scaling or lichenification: 4 points if the area is larger than 75% of the exposed skin area; 3 points if the area is between 40% and 75%; 2 points if the area is between 10% and 40%; 1 point if the area is less than 10%; 0 points if there is no bleeding or erythema.

[0099] like Figure 3A , Figure 3B As shown, the skin of mice in the normal group was smooth and free of dermatitis, with a dermatitis score of 0. The dermatitis score of the model group was relatively high (P<0.05). Erythema and edema appeared on the back skin. The dermatitis scores of mice in the dexamethasone group and the OSHP prescription combination group were lower than those in the model group. The erythema on the back of the mice was reduced, the desquamation was improved, and the area of ​​scabs gradually shrank. The OSHP prescription combination group and the dexamethasone group had a good therapeutic effect on atopic dermatitis in mice. The back skin of the mice did not show redness and swelling, but only small patches of erythema and scaling. Compared with the model group, the symptoms of mice in the treatment group were significantly improved, proving that the OSHP prescription combination has a good therapeutic effect on atopic dermatitis.

[0100] 4. Changes in the scores of behavioral indicators (scratching times) of AD mice

[0101] The scratching behavior of AD mice is also an important indicator of the severity of mouse dermatitis. The observation method is to place them alone in a transparent mouse cage ten minutes after DNCB sensitization and observe the scratching behavior of the mice on the back and ears. A series of one or more scratching movements of the mouse's hind paws toward the ears is counted as one scratch, and the scratch ends when the mouse's hind paws are placed back on the floor. The timer is 10 minutes, and the data is recorded and the average value is finally taken.

[0102] like Figure 3C As shown, the mice in the normal group had no scratching behavior because their skin was smooth without sensitization. The model group showed obvious scratching behavior with a very high frequency, indicating that the mice had a high degree of skin inflammation. The dexamethasone group and the OSHP capsule group scratched less frequently than the model group and showed significant improvement.

[0103] 5. Changes of spleen index in AD mice

[0104] The mice were killed by dislocation and the spleen was taken out and placed on filter paper for weighing. After weighing, the spleen index was calculated = spleen mass (mg) / mouse body weight (mg). The larger the spleen coefficient, the more severe the inflammatory response of the mouse. Therefore, the spleen index is also an indicator of the severity of mouse specific dermatitis.

[0105] like Figure 3D , Figure 3E As shown, the spleen index of the OSHP prescription combination group and the dexamethasone group was lower than that of the model group mice, indicating that the OSHP prescription combination can reduce the inflammation of mice and play a role in the treatment of atopic dermatitis.

[0106] 6. Pathological changes of skin tissue in AD area of ​​mice

[0107] On the 13th day of the experiment, the mice were killed by dislocation, and the damaged skin on the back was removed and fixed with 4% paraformaldehyde, routinely dehydrated, paraffin-embedded, sectioned, and stained with hematoxylin-eosin (HE). The pathological changes of the back skin tissue were observed under an optical microscope.

[0108] like Figure 4A , Figure 4B , Figure 4C , Figure 4D As shown in the figure, the back skin tissue of the mice in the normal group was normal without obvious changes, the structure of each layer of the skin was intact, the dermis and epidermis were clearly demarcated, and no inflammatory cell infiltration was found. The back skin of the mice in the model group showed severe hyperkeratosis, thickening of the acanthus layer, and a large number of inflammatory cell infiltrations in the upper dermis, which fully met the characteristics of AD. Compared with the model group, the pathological tissue results of the mice in the OSHP prescription combination group showed that the phenomenon of epidermal keratinization was reduced and the severity of dermatitis lesions was reduced.

[0109] 7. Effects of OSHP prescription combination on cytokines in AD skin lesions

[0110] At the end of the experiment, when we were collecting mouse tissues, we collected blood from the mice's eyes, centrifuged it, and put it in a -80-degree refrigerator. We then used ELISA to detect the blood concentrations of Ig E and IL-17 in the mouse serum. We compared the concentrations of Ig E and IL-17 between different groups to compare the degree of inflammation in the mice, and measured the therapeutic effect of the OSHP prescription combination on atopic dermatitis by measuring the concentrations of immune factors in the serum. This experiment mainly used Mouse Immunoglobulin E (IgE) ELISA Kit and Mouse IL-17 ELISA Kit to detect the expression levels of Ig E and IL-17, respectively.

[0111] like Figure 5A , Figure 5B As shown, compared with the model group, after treatment, the expression levels of Ig E and IL-17 in the mice of the OSHP prescription combination group and the positive control dexamethasone group were significantly lower than those in the model group. Therefore, the OSHP prescription combination may reduce the release of inflammatory factors in the process of atopic dermatitis through the effect of immune regulation, so as to achieve the effect of reducing inflammation and relieving dermatitis. Compared with the positive control, it can be seen that the OSHP prescription combination and dexamethasone have basically the same effect in relieving inflammation. Therefore, the OSHP prescription combination can treat AD by reducing the expression of inflammatory factors such as Ig E and IL-17 in the serum during inflammation.

[0112] like Fig. 6A As shown in Figure 2, the absolute number of lymphocytes in the peripheral blood of mice in the model group was significantly higher than that in normal mice. Fig.6D Compared with the model group, the absolute number of lymphocytes in the peripheral blood of the dexamethasone-treated group increased significantly, such as Fig. 6A As shown in Figure 2, the OSHP prescription combination group tended to suppress the proportion and absolute number of lymphocytes in peripheral blood compared with the model group, as shown in Figure 2 Fig.6D It is noteworthy that when comparing the OSHP prescription combination treatment group with the dexamethasone treatment group, there were statistically significant differences in the proportion and absolute number of suppressed lymphocytes, as shown in Figure 2. Fig. 6A and 6D Secondly, compared with the normal group mice, the proportion and absolute number of monocytes and neutrophils in the peripheral blood of the model group mice showed an upward trend, as shown in Figure 6B , Figure 6C , Fig. 6E and Fig. 6F As shown, the dexamethasone treatment group showed an inhibitory trend in the absolute number and proportion of monocytes and granulocytes (the average change was more obvious, but no significant statistical difference was found due to the large intra-group difference). The OSHP prescription combination group had no effect on the proportion and absolute number of monocytes and granulocytes in mice. Since lymphocytes are the main cell type in peripheral blood, accounting for 70-90% of peripheral blood monocytes, it can be seen that the OSHP prescription combination group mainly inhibits AD by inhibiting the proportion and absolute number of lymphocytes and the cytokines they secrete.

[0113] The above experimental results show that the OSHP prescription combination has a significant improvement on the treatment of AD at a concentration of 50 mg / kg. The research results show that the OSHP prescription combination has an improvement effect on AD, can reduce the epidermal hyperplasia of AD mice, and play a role in repairing the skin of mice. In addition, the H2 released by an oyster shell hydrogen powder prescription combination with a buffered pH value in the mouse body can also play an anti-inflammatory role, reduce the release of inflammatory factors, and alleviate the damaged skin on the back of AD mice.

[0114] Experiment 2: Experimental study on the effect of the composition of Example 1 on promoting wound healing in scalded mice

[0115] Scald is an injury caused by high-temperature liquid, high-temperature solid or high-temperature steam. The degree of scald is divided into three degrees. First-degree scald only damages the surface of the skin, with mild redness and swelling without blisters; second-degree scald damages the dermis, with blisters of varying sizes; third-degree scald is a subcutaneous injury involving fat, muscle layer and bone. At present, small-area mild burns should be rinsed with room temperature water for a few minutes at the first time, and then the affected area should be disinfected and bandaged; small-area deep burns should be disinfected and bandaged after disinfection and bandaging to promote wound recovery according to the actual condition; large-area burns need to be supplemented with fluid within 24 hours after medical treatment, maintain the patient's body fluid balance, keep the wound clean, change the dressing bandage on time, etc., and perform one or more skin grafting operations depending on the degree of burns and recovery. Example 2 The improvement of the OSHP prescription combination on scalded mice was evaluated by establishing a scalded mouse model, providing data support for the application of an OSHP prescription combination with a buffered pH value in functional products.

[0116] 1. Reagent Preparation

[0117] Preparation of 2% sodium alginate: 1 g of sodium alginate was dissolved in 50 ml of water and placed on a magnetic stirrer to dissolve.

[0118] OSHP prescription combination gel: effective concentration 100mg / ml, dissolved with 2% sodium alginate.

[0119] 2. Animal burn modeling, grouping and medication

[0120] The experiment was divided into three groups: model group, empty load group (gel was applied only on the affected area), and OSHP prescription combination group. Use an electric shaver to shave 2.5cm×2.5cm of the mouse back skin, remove the mice with back skin problems to avoid interference, and use depilatory cream to remove hair twice on the shaved area to ensure that the mouse back skin can be completely exposed. After using the depilatory cream, wash with water to prevent the depilatory cream from corroding the mouse skin and affecting the modeling effect. On the first day of the experiment, all three groups of mice were modeled under anesthesia using a scalding instrument. Drug treatment was started on the day of modeling, and the drug concentration of the OSHP prescription combination group was 100mg / ml. The medication was administered externally to ensure that the drug efficacy could fully exert its effect. The medication was administered for a total of 26 days, and the experimental materials were collected on the 26th day.

[0121] 3. Healing of the back wound of scalded mice

[0122] In this experiment, the wound surface of mice was photographed and recorded every other day to compare the surface healing effect. The efficacy of a prescription combination of oyster shell hydrogen powder with a buffered pH value was judged based on the changes in the healing status of the mouse wound surface.

[0123] like Figure 7 As shown, the OSHP prescription combination group had a significant improvement in the treatment of scalded mouse wounds at a concentration of 100 mg / ml. The wounds of mice in the model group and the empty group were completely healed on the 20th day, and the wounds of mice in the OSHP prescription combination group were completely healed on the 14th day. The wound healing time was one week shorter than that of other groups. This may be related to the hydrogen released by OSHP in the wounds of mice, which reduces the release of inflammatory factors, accelerates the healing of scalded wounds in mice, repairs mouse skin, and promotes skin growth and healing. It is proved that an OSHP prescription combination with a buffered pH value can quickly promote wound healing.

[0124] 4. Mouse weight changes

[0125] In this experiment, the mice were weighed every day. The effect of an OSHP prescription combination with a buffered pH value on the wound healing of scalded mice was determined based on the changes in the mice's weight.

[0126] Mouse weight is an important indicator for evaluating mouse growth and development, as well as an evaluation indicator for mouse quality of life and disease recovery. Figure 8 As shown in the figure, the weight gain curve of scalded mice showed that the weight gain trend of mice in the OSHP prescription combination group was better than that in other groups. This may be related to the faster wound healing speed of mice in the OSHP prescription combination group, better disease recovery of mice, and improved quality of life due to reduced pain caused by disease recovery.

[0127] The oyster shell hydrogen powder composition with pH buffering function of the present invention has significant anti-inflammatory and wound healing effects, and can provide hydrogen to the body stably for a long time. By taking the composition of the present invention orally, the H2 content in the human body can be effectively increased, thereby playing an anti-inflammatory and wound growth and healing role. The composition of the present invention also has the advantages of high safety and small side effects, and is suitable for long-term use by a large number of people.

[0128] The present invention utilizes solid-state hydrogenation technology to produce a hydrogen-rich OSHP, which has the characteristic of slowly and continuously releasing hydrogen when in contact with water. After external application, it can slowly and continuously release hydrogen on the body surface to promote rapid healing of wounds.

[0129] A H2-rich oyster shell powder produced by solid-state hydrogenation technology has the property of slowly and continuously releasing H2 when it comes into contact with water. Oral administration through the gastrointestinal tract can slowly and continuously release H2 with certain health benefits, thereby achieving significant anti-inflammatory, immune regulation, and metabolic regulation effects.

[0130] The present invention adopts OSHP, citric acid, hydroxyethyl methyl cellulose (HEMC) and starch as the contents of an OSHP prescription combination with a buffered pH value, utilizes the characteristic of OSHP that can release H2 for a long time, adds citric acid (CA) for acid-base buffering, and adjusts the pH value of an OSHP prescription combination for anti-inflammation, regulating immunity, regulating metabolism and promoting rapid wound healing to 7. Hydroxyethyl methyl cellulose (HEMC) is then added to improve the stability of the drug and increase the sustained release effect of the drug, thereby achieving the purpose of enhancing the safety of the drug.

Claims

1. An oyster shell hydrogen powder composition having a pH buffering function, characterized in that: The invention is composed of the following components by mass percentage: 40% to 50% oyster shell hydrogen powder, 20% to 30% starch, 10% to 15% citric acid, and 15% to 20% hydroxyethyl methyl cellulose, and the sum of the percentages of the above components is 100%.

2. The oyster shell hydrogen powder composition with pH buffering function according to claim 1, characterized in that: The invention is composed of the following components by mass percentage: 45% oyster shell hydrogen powder, 24% starch, 13% citric acid and 18% hydroxyethyl methyl cellulose.

3. The oyster shell hydrogen powder composition with pH buffering function according to claim 1, characterized in that: The citric acid acts as an acidity regulator to adjust the pH value of the composition to a stable neutral state.

4. The oyster shell hydrogen powder composition with pH buffering function according to claim 1, characterized in that: The hydroxyethyl methylcellulose is used as a sustained-release preparation to improve the stability of the composition and enhance the safety of the composition.

5. The oyster shell hydrogen powder composition with pH buffering function according to any one of claims 1 to 4, characterized in that: The oyster shell hydrogen powder composition is packaged in capsule shells, the capsule shells are made of edible and easily digestible gelatin, and are made into capsules with a specification of 0.45 g of content per capsule.

6. The method for preparing the oyster shell hydrogen powder composition having pH buffering function according to claim 5, characterized in that: Prepare according to the following steps: Step 1, sieve oyster shell hydrogen powder, starch, citric acid and hydroxyethyl methylcellulose through a 100-mesh sieve for later use; Step 2: According to the formula, accurately weigh oyster shell hydrogen powder, starch, citric acid, and hydroxyethyl methylcellulose and mix them; Step 3, filling the total mixed material into hollow capsules; Step 4: Pack the filled capsules.

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