A repair functional composition HAmella containing sodium hyaluronate and tremella polysaccharide

Abstract

The present invention discloses a repair functional composition HAmella containing sodium hyaluronate and tremella polysaccharide, belonging to the field of biomedical technology. The preparation method includes: mixing sodium hyaluronate and tremella polysaccharide, dissolving them in water to prepare a polysaccharide aqueous solution; wherein, the molecular weight of the sodium hyaluronate is 100 - 300 KD, and the molecular weight of the tremella polysaccharide is 50 - 200 KD; dissolving saponin in water to prepare an aqueous phase, and then adding camellia oil to the aqueous phase to prepare a camellia oil microemulsion; mixing the polysaccharide aqueous solution and the camellia oil microemulsion evenly, and spray drying to obtain a gastric mucosa protectant. The preparation method of the protectant of the present invention is simple, shows excellent performance in gastrointestinal protection, and has an injury inhibition rate of about 50% in an ethanol-induced gastric ulcer model.

Description

Technical Field

[0001] The present invention relates to a repair functional composition HAmella containing sodium hyaluronate and tremella polysaccharide, belonging to the field of biomedical technology. Background Art

[0002] Gastric mucosal injury is a common digestive disease with a high recurrence rate and is difficult to completely cure, afflicting millions of people worldwide. Excessive alcohol consumption, long-term use of non-steroidal anti-inflammatory drugs, excessive stress, and Helicobacter pylori infection can all lead to gastric mucosal injury. Exploring safer and more effective gastric protection products or finding raw materials with auxiliary protection functions to protect gastric mucosal injury has attracted much attention.

[0003] Microemulsion is a homogeneous, low-viscosity, thermodynamically stable transparent or semi-transparent dispersion system spontaneously formed by surfactants, co-surfactants, oil, and water in appropriate proportions. Camellia oil is a type of high-quality edible oil. Numerous studies have shown that camellia oil has health care functions such as antioxidant, immune regulation, and gastrointestinal regulation. Preparing camellia oil into an oral microemulsion can not only improve the greasy taste of camellia oil but also solve the problem of its easy oxidation.

[0004] Polysaccharides, such as sodium hyaluronate and tremella polysaccharide, can combine with the surface of the gastric mucosa to form a mucosal barrier, reducing the secretion of gastric acid and pepsin or the inflammatory reaction of the gastric mucosa. Therefore, they are often used to prepare gastrointestinal protection products.

[0005] Tea saponin is a type of natural glycoside compound, mainly a mixture of oleanane-type pentacyclic triterpenoid saponins. Its basic structure consists of three parts: a hydrophobic aglycone, a hydrophilic sugar body, and an organic acid. Studies have shown that tea saponin itself has obvious anti-leakage and anti-inflammatory functions.

[0006] The polysaccharide structure is very complex. Its complexity not only refers to the chemical structure but mainly to the variability of the molecular conformation. For example, polysaccharide molecules can have a coil structure, a single-chain structure, a double helix structure, a triple helix structure, etc., and there are unique interactions between these structures, especially secondary bond interactions such as hydrogen bond, hydrophobic interaction, electrostatic interaction, and van der Waals force. Many studies have confirmed that the physiological activity of polysaccharides is closely related to their structure. Even the loss of activity will occur after the stereostructure of some polysaccharides is changed. However, the higher-order structure of polysaccharides is not immutable. Changes in many factors such as polysaccharide molecular weight, polysaccharide concentration, monosaccharide composition, and pH value can lead to changes in the stereoconformation and asymmetry of polysaccharides in solution, thereby causing changes in activity. In the O / W type camellia oil microemulsion prepared with tea saponin, the hydrophilic sugar body bond in the molecular structure of tea saponin is in the external aqueous phase and may non-covalently bind to the polysaccharide molecules in the aqueous phase. The present invention unexpectedly found in the study that the microemulsion system obtained by adding sodium hyaluronate and tremella polysaccharide with a specific molecular weight to this microemulsion system has a more excellent gastrointestinal protection effect. Summary of the Invention

[0007] To solve the above problems, the present invention provides a gastrointestinal protection product containing sodium hyaluronate and tremella polysaccharide and a preparation method thereof, and the product has the remarkable characteristic of protecting the gastrointestinal tract.

[0008] The first object of the present invention is to provide a preparation method of a repair functional composition HAmella containing sodium hyaluronate and tremella polysaccharide, comprising the following steps:

[0009] S1. Mix sodium hyaluronate and tremella polysaccharide, dissolve them in water, and prepare a polysaccharide aqueous solution; wherein, the molecular weight of the sodium hyaluronate is 100 - 300KD, and the molecular weight of the tremella polysaccharide is 50 - 200KD;

[0010] S2. Dissolve tea saponin in water to prepare an aqueous phase, and then add camellia oil to the aqueous phase to prepare a camellia oil microemulsion;

[0011] S3. Mix the polysaccharide aqueous solution obtained in S1 and the camellia oil microemulsion obtained in S2 evenly, and perform spray drying to obtain the repair functional composition.

[0012] Further, in step S1, the mass ratio of tremella polysaccharide to sodium hyaluronate is 5:1 - 1:5.

[0013] Further, in step S1, the concentration of the polysaccharide aqueous solution is 0.05 - 0.15 g / 100 mL.

[0014] Further, in step S2, the mass ratio of tea saponin to the volume of water is 1 - 5 g / 100 mL.

[0015] Further, in step S2, the mass ratio of camellia oil to the aqueous phase is 1:50 - 1:10.

[0016] Further, in step S2, the camellia oil microemulsion is prepared by high - speed shearing followed by high - pressure homogenization.

[0017] Further, the high - speed means the rotation speed ≥ 10000 rpm.

[0018] Further, the high - pressure means the pressure is 100 - 120 psi.

[0019] Further, in step S3, the mass ratio of the camellia oil microemulsion to the polysaccharide aqueous solution is 1:30 - 1:60.

[0020] Further, in step S3, the inlet air temperature of the spray drying is 180 - 220 °C, the outlet air temperature is 60 - 90 °C, and the flow rate is 5 - 10 mL / min.

[0021] The second object of the present invention is to provide a repair functional composition obtained by the above preparation method.

[0022] The third object of the present invention is to provide the application of the above repair functional composition in the preparation of drugs for gastrointestinal diseases, such as drugs for treating gastric ulcers.

[0023] Furthermore, the drug for gastrointestinal diseases is a drug for treating gastric mucosal injury.

[0024] Advantages of the present invention:

[0025] The present invention uses tea saponin to prepare an O / W type tea oil microemulsion. The hydrophilic glycoside bond in the molecular structure of tea saponin is in the outer aqueous phase and may form a non-covalent bond with the polysaccharide molecules in the aqueous phase. The present invention unexpectedly discovers in the research that the microemulsion system obtained by adding sodium hyaluronate with a specific molecular weight and tremella polysaccharide in this microemulsion system has a more excellent gastrointestinal protection effect. The injury inhibition rate is about 50% in the ethanol-induced gastric ulcer model, and the preparation method is simple and convenient for industrialization. Specific embodiments

[0026] The following combines specific embodiments to further illustrate the present invention, so that those skilled in the art can better understand the present invention and be able to implement it, but the exemplified embodiments do not limit the present invention.

[0027] Example 1

[0028] Take sodium hyaluronate (purchased from Shandong Focus Frida Biotechnology Co., Ltd.) and 50KD tremella polysaccharide (purchased from Shandong Focus Frida Biotechnology Co., Ltd.) powders, mix them evenly according to a weight ratio of 1:1, and configure the mixed powder into a first aqueous solution at a concentration of 0.1g / 100ml; dissolve tea saponin (purchased from Guizhou Camellia Comprehensive Development Co., Ltd.) in water according to a ratio of 2g / 100mL, and stir evenly; mix camellia oil (purchased from Guizhou Camellia Comprehensive Development Co., Ltd.) and the tea saponin solution evenly according to a ratio of 1:50, shear at a high speed of 1000rpm, and subject the obtained solution to high-pressure homogenization at a pressure of 100psi; mix the obtained solution and the first aqueous solution evenly according to a weight ratio of 1:50 and then spray dry, with an inlet air temperature of 180°C, an outlet air temperature of 90°C, and a flow rate of 6ml / min.

[0029] In the above steps, sodium hyaluronate refers to sodium hyaluronate with different molecular weights.

[0030] Table 1 Composition of different sodium hyaluronate formulations

[0031] Number Average molecular weight (kd) A 5 B 100 C 150 D 300 E 500

[0032] Raw materials A - E prepared with different sodium hyaluronates were used. Referring to the health product evaluation method, an ethanol-induced gastric mucosal injury rat model was used as the evaluation method. The gastric ulcer index and injury inhibition rate observed macroscopically and histologically in tissues were used as evaluation indicators to study and compare the gastric mucosal protective effects of different raw materials. The specific method was as follows: SD rats were adaptively fed for one week and randomly divided into a blank group, a model group, and an experimental group. The blank and model groups were intragastrically administered distilled water every day; the experimental group was intragastrically administered different raw materials at a dose of 100 mg / kg BW once a day for 21 consecutive days. After the last intragastric administration, the rats were fasted and allowed free access to water for 12 h. The model group and the experimental group were intragastrically administered absolute ethanol (10 mL / kg), and the blank group was not intragastrically administered. After 1 h, the rats were sacrificed, and the gastric tissues were taken out. The occurrence of ulcers in the gastric mucosal layer was carefully observed, and the cumulative score of the length of erosions, ulcers, bleeding, etc. limited to the gastric epithelium was used, and the length and width of the ulcer stripes were measured with a ruler. Based on the following scoring criteria: normal gastric mucosa was 0 points; the presence of spotted perforations was counted as 1 point; bleeding length > 4 mm was counted as 5 points, and the sum of each score was the total injury index score of the animal. The injury index was the average value of the total injury index scores of the animals in that group. The injury inhibition rate was calculated according to the following formula:

[0033] Injury inhibition rate = (A0 - A1) / A0

[0034] where A0 was the gastric ulcer index of the model group and A1 was the gastric ulcer index of the experimental group. The closer the inhibition rate was to 1, the stronger the gastrointestinal protective effect.

[0035] Group Injury inhibition rate A 15％ B 35％ C 45％ D 33％ E 16％

[0036] Example 2

[0037] 150 kd sodium hyaluronate and tremella polysaccharide powder were mixed evenly at a weight ratio of 1:1, and the mixed powder was prepared into a No. 1 aqueous solution at a concentration of 0.1 g / 100 ml; tea saponin was dissolved in water at a ratio of 2 g / 100 mL and stirred evenly; camellia oil and the tea saponin solution were mixed evenly at a ratio of 1:50 and sheared at a high speed of 1000 rpm, and the obtained solution was homogenized under high pressure at 100 psi; the obtained solution was mixed evenly with the No. 1 aqueous solution at a weight ratio of 1:50 and then spray-dried, with an inlet air temperature of 180 °C, an outlet air temperature of 90 °C, and a flow rate of 6 ml / min.

[0038] In the above steps, tremella polysaccharide refers to tremella polysaccharides with different molecular weights.

[0039] Table 2 Composition of different tremella polysaccharide formulations

[0040] Number Molecular weight (kd) F 35 G 50 H 120 I 200 J 800

[0041] Raw materials F - J prepared from different tremella polysaccharides were used as raw materials for animal experiments according to the method in Example 1, and the injury inhibition rate was measured.

[0042]

[0043]

[0044] Example 3

[0045] 150 kd sodium hyaluronate and 100 kd tremella polysaccharide powder were mixed evenly at a weight ratio of 5:1, and the mixed powder was prepared into a first aqueous solution at a concentration of 0.1 g / 100 ml; the surfactant was dissolved in water at 2 g / 100 mL and stirred evenly; camellia oil and tea saponin solution were mixed evenly at a ratio of 1:50, sheared at a high speed of 1000 rpm, and the obtained solution was homogenized under high pressure at 100 psi; the obtained solution was mixed evenly with the first aqueous solution at a weight ratio of 1:50 and then spray-dried, with an inlet air temperature of 180 °C, an outlet air temperature of 90 °C, and a flow rate of 6 ml / min.

[0046] The surfactant includes the following:

[0047] Number Type K Tea saponin L Sodium dodecyl sulfonate M Fatty alcohol polyoxyethylene ether N Disodium monolauryl sulfosuccinate

[0048] Raw materials K - N prepared from different materials were used as raw materials for animal experiments according to the method in Example 1, and the injury inhibition rate was measured.

[0049] Group Inhibition rate K 43％ L 13％ M 12％ N 14％

[0050] Example 4

[0051] 150 kd sodium hyaluronate and 100 kd tremella polysaccharide powder were mixed evenly at a weight ratio of 1:1, and the mixed powder was prepared into a first aqueous solution at a concentration of 0.1 g / 100 ml; tea saponin was dissolved in water at 2 g / 100 mL and stirred evenly; the oil raw material and tea saponin solution were mixed evenly at a ratio of 1:50, sheared at a high speed of 1000 rpm, and the obtained solution was homogenized under high pressure at 100 psi; the obtained solution was mixed evenly with the first aqueous solution at a weight ratio of 1:50 and then spray-dried, with an inlet air temperature of 180 °C, an outlet air temperature of 90 °C, and a flow rate of 6 ml / min.

[0052] The oil raw material specifically includes the following:

[0053] Number Type O Camellia oil P Rapeseed oil Q Soybean oil R Corn oil

[0054] O - R raw materials were prepared from different materials. Referring to the method in Example 1, animal experiments were carried out using raw materials O - R as raw materials, and the injury inhibition rate was measured.

[0055] Group Inhibition rate O 46％ P 14％ Q 11％ R 13％

[0056] Example 5

[0057] 150 kd sodium hyaluronate and 100 kd tremella polysaccharide powder were mixed evenly according to a weight ratio of 5:1. The mixed powder was prepared into a first aqueous solution at a concentration of 0.1 g / 100 ml; tea saponin was dissolved in water at 2 g / 100 mL and stirred evenly; the oil raw material and the tea saponin solution were mixed evenly according to a ratio of 1:50, and high - speed shearing was carried out at 1000 rpm. The obtained solution was homogenized under high pressure at 100 psi; the obtained solution and the first aqueous solution were mixed evenly according to different weight ratios and then spray - dried. The inlet air temperature was 180 °C, the outlet air temperature was 90 °C, and the flow rate was 6 ml / min.

[0058] The different weight ratios are shown in the following table:

[0059] Number Ratio S 1：200 T 1：30 U 1：60 V 1：10

[0060] Different raw materials S - V were obtained in different proportions. Referring to the method in Example 1, animal experiments were carried out using raw materials S - V as raw materials, and the injury inhibition rate was measured.

[0061] Group Inhibition rate S 12％ T 47％ U 42％ V 11％

[0062] Comparative Example 1

[0063] 150 kd sodium hyaluronate was prepared into a first aqueous solution at a concentration of 0.1 g / 100 ml; tea saponin was dissolved in water at 2 g / 100 mL and stirred evenly; camellia oil and the tea saponin solution were mixed evenly according to a ratio of 1:50, and high - speed shearing was carried out at 1000 rpm. The obtained solution was homogenized under high pressure at 100 psi; the obtained solution and the first aqueous solution were mixed evenly according to a weight ratio of 1:50 and then spray - dried. The inlet air temperature was 180 °C, the outlet air temperature was 90 °C, and the flow rate was 6 ml / min.

[0064] Comparative Example 2

[0065] Take 100 kd tremella polysaccharide powder and prepare it into the first aqueous solution at a concentration of 0.1 g / 100 ml; dissolve tea saponin in water at a ratio of 2 g / 100 mL and stir evenly; mix camellia oil and the tea saponin solution evenly at a ratio of 1:50, and perform high-speed shearing at 1000 rpm. The obtained solution is subjected to high-pressure homogenization at a pressure of 100 psi; the obtained solution is mixed evenly with the first aqueous solution at a weight ratio of 1:50 and then spray-dried. The inlet air temperature is 180 °C, the outlet air temperature is 90 °C, and the flow rate is 6 ml / min.

[0066] Comparative Example 3

[0067] According to the ratio in Example 1, take 150 kd sodium hyaluronate, 100 kd tremella polysaccharide, camellia oil, and tea saponin, stir and mix them, and then spray-dry.

[0068] Compare the injury inhibition rates of Example 1 and Comparative Examples 1 to 3. At the same time, take gastric mucus to measure the activity of pepsin (gastric mucosal protein is a proteolytic enzyme that has the function of decomposing proteins and is one of the attack factors causing gastric mucosal damage. Therefore, reducing the activity of pepsin is an important aspect of anti-gastric mucosal damage).

[0069] Raw material Injury inhibition rate Pepsin activity Example 1 56％ 210U / ml Comparative example 1 14％ 232U / ml Comparative example 2 13％ 234U / ml Comparative example 3 11％ 240U / ml

[0070] The above-described embodiments are merely preferred embodiments given to fully illustrate the present invention, and the protection scope of the present invention is not limited thereto. Equivalent substitutions or transformations made by those skilled in the art on the basis of the present invention are all within the protection scope of the present invention. The protection scope of the present invention shall be subject to the claims.

Claims

1. A preparation method of a repair function composition containing sodium hyaluronate and tremella polysaccharide for treating gastric mucosal injury, characterized in that, It includes the following steps: S1. Mix sodium hyaluronate and tremella polysaccharide, dissolve them in water to prepare a polysaccharide aqueous solution; wherein, the molecular weight of the sodium hyaluronate is 100 - 300 KD, and the molecular weight of the tremella polysaccharide is 50 - 200 KD; S2. Dissolve tea saponin in water to prepare an aqueous phase, and then add camellia oil to the aqueous phase to prepare a camellia oil microemulsion; S3. Mix the polysaccharide aqueous solution obtained in S1 and the camellia oil microemulsion obtained in S2 evenly, and perform spray drying to obtain the repair functional composition; In step S1, the mass ratio of tremella polysaccharide to sodium hyaluronate is 5:1 - 1:5, and the concentration of the polysaccharide aqueous solution is 0.05 - 0.15 g / 100mL, In step S2, the mass - to - volume ratio of tea saponin to water is 1 - 5 g / 100mL, and the mass ratio of camellia oil to the aqueous phase is 1:50 - 1:10, In step S3, the mass ratio of the camellia oil microemulsion to the polysaccharide aqueous solution is 1:30 - 1:

60.

2. The preparation method according to claim 1, characterized in that: In step S3, the inlet air temperature for spray drying is 180 - 220°C, and the outlet air temperature is 60 - 90°C.

3. The repair functional composition obtained by the preparation method according to claim 1 or 2.

4. Use of the repair functional composition according to claim 3 in the preparation of drugs for gastrointestinal diseases, characterized in that: The gastrointestinal disease drug is a drug for treating gastric mucosal injury.

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