Composition capable of protecting gastric mucosa as well as preparation method and application of composition

Abstract

The invention particularly discloses a dendrobium officinale polysaccharide composition with a gastric mucosa protection effect and a preparation method and application thereof, the composition is prepared on the basis of dendrobium officinale polysaccharide and a Brazilian ginseng extract, and the mass ratio of the Brazilian ginseng extract to the dendrobium officinale polysaccharide is (1-2): (3-4). The composition prepared by synergistic compatibility of the dendrobium officinale polysaccharide and the Brazilian ginseng extract has a good gastric mucosa protection effect.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and specifically relates to a composition for protecting gastric mucosa, a preparation method and an application thereof. Background Art

[0002] With the awakening of people's health awareness, residents have increased their yearning for a healthy life, and the number of people with stomach diseases has gradually increased. Scientific stomach care is urgent. The main means of stomach care are diet and medicine. The use of diet and nutritional intervention and regulation has become an important and economical measure. The stomach is one of the important organs of the human body. Its main physiological function is to receive and decompose water and grain, which is the basic guarantee for normal human life. It plays an important role in the healthy development of human body and mind. An unhealthy lifestyle often causes damage to the gastric mucosa, which will cause various stomach diseases in the long run. Therefore, for the increasing number of sub-healthy people with gastric mucosal damage, it is of positive significance to provide them with a composition that promotes the repair of gastric mucosal damage.

[0003] In recent years, natural polysaccharides from plants have been proven to have many physiological effects such as anti-oxidation and anti-inflammatory. There are also many studies on the protection of gastric mucosal damage by these polysaccharides, and the effect is remarkable. Dendrobium candidum can nourish the stomach, and many studies have shown that Dendrobium candidum polysaccharides have the effect of restoring and protecting gastric mucosal damage in various models. Brazilian ginseng is native to South America and can invigorate qi and strengthen the spleen. Modern research shows that it mainly contains triterpenes, triterpenoid saponins, steroids and other chemical components, and has pharmacological activities such as anti-tumor and anti-inflammatory. Summary of the invention

[0004] The present invention is to solve at least one of the technical problems existing in the prior art. The first purpose of the present invention is to provide a composition for protecting gastric mucosa.

[0005] The second purpose is to provide a method for preparing a composition for protecting gastric mucosa, which has simple process, convenient operation and is suitable for large-scale industrial production.

[0006] The third purpose is to provide an application of a composition for protecting gastric mucosa.

[0007] To achieve the above object, the present invention provides the following solutions:

[0008] The invention provides a composition with the function of protecting gastric mucosa. The composition is prepared based on dendrobium officinale polysaccharide and Brazilian ginseng extract, and the mass ratio of the Brazilian ginseng extract to the dendrobium officinale polysaccharide is 1:2:3-4.

[0009] A method for preparing a composition for protecting gastric mucosa, comprising:

[0010] Preparation of Brazilian ginseng extract: weigh Brazilian ginseng root powder with a mass ratio of 1:1-1:3, add ethanol solution and reflux extract for several times, collect and combine the extract after filtering, and concentrate it in a rotary evaporator to an extract with a solid-liquid ratio relative density of 1.0-1.2, so as to obtain Brazilian ginseng extract;

[0011] Preparation of Dendrobium officinale polysaccharide:

[0012] After extracting the Dendrobium officinale with water and precipitating it with alcohol, Sevage reagent was used to remove protein to obtain a crude polysaccharide solution, and water and organic solvent were rotary evaporated and freeze-dried to obtain Dendrobium officinale polysaccharide;

[0013] The Dendrobium officinale polysaccharide and the Brazilian ginseng extract are fully mixed to obtain the composition.

[0014] Furthermore, before the water extraction and alcohol precipitation, the method further comprises crushing the Dendrobium officinale, mixing and soaking the mixture with the ethanol solution, and centrifuging the mixture to obtain a first precipitate;

[0015] Furthermore, the water extraction and alcohol precipitation comprises mixing the first precipitate with water, heating, centrifuging, taking the supernatant to obtain a total polysaccharide extract; mixing the total polysaccharide extract with an ethanol aqueous solution with a volume fraction of 80%, standing, and obtaining a second precipitate.

[0016] Preferably, the volume ratio of the crushed Dendrobium officinale to the ethanol solution is 1 g: (15-20) mL.

[0017] Preferably, the volume ratio of the first precipitate to water is 1 g: (15-20) mL.

[0018] Preferably, the heating treatment is carried out at 95-100°C for 2-3 hours and the extraction is performed 2-3 times.

[0019] Preferably, the volume ratio of the total polysaccharide extract and the ethanol aqueous solution is 1:(3-4).

[0020] The present invention also provides an application of a composition having a gastric mucosa protecting function in preparing a product for nourishing the stomach.

[0021] According to the theory of traditional Chinese medicine, the present invention compounds Dendrobium candidum polysaccharide and Brazilian ginseng extract into a composition, which has a significant effect on the protection of gastric mucosal damage after being matched in a certain proportion. Animal experiments show that the composition disclosed by the present invention can reduce the content of TNF-α, IL-6, PG-I, PG-II and GAS in the serum of rats with ethanol-induced gastric mucosal damage model, and has a protective effect on gastric mucosal damage. The composition of the present invention is of great significance for the development of health products for auxiliary protection of gastric mucosal damage and drugs for treating gastric mucosal damage.

[0022] The functions of various Chinese medicines in the technical solution of the present invention are:

[0023] Dendrobium officinale: The dried stem of Dendrobium officinale, an orchid plant, has multiple biological properties such as antioxidant, anti-inflammatory and immunomodulatory, and has multiple health benefits such as anti-cancer, anti-diabetic, cardiovascular protection, gastrointestinal regulation, liver protection, lung protection and neuroprotection.

[0024] Brazilian ginseng: The dried root of Brazilian ginseng, a herbaceous plant of the genus Phaphiaceae in the Amaranthaceae family, is widely used to treat chronic fatigue syndrome, immune disorders, blood circulation, inflammation, asthma, hyperglycemia, hypertension and other diseases. It has high health and medicinal value. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 It is the fitting curve diagram of Example 1 and Example 2.

[0026] Figure 2 These are cell pictures of each group in Example 3 and Example 4, A: blank group, B: model group, C: positive group, D: polysaccharide group; E: Brazilian ginseng group, F: composition a, G: composition b.

[0027] Figure 3 This is a comparison chart of TNF-α, IL-β, PG-I, PG-II, and GAS levels in each group of Example 3 and Example 4. DETAILED DESCRIPTION

[0028] In order to make the purpose and advantages of the present invention more understandable, the present invention is further explained below in conjunction with specific embodiments, but the specific embodiments do not limit the present invention in any way, and those skilled in the art can make similar generalizations without violating the connotation of the present invention.

[0029] Example 1 Preparation of Dendrobium officinale polysaccharide

[0030] After the Dendrobium candidum was crushed by a grinder, 15 times the volume of ethanol solution was added, stirred evenly, and soaked at room temperature for 12 hours. After centrifugation, the supernatant was discarded, the powder was dried, 15 times the volume of water was added, and heated at 95°C for 2 hours. After cooling to room temperature, the supernatant was centrifuged. Repeat the extraction twice, combine the supernatants, and obtain the total polysaccharide extract. Add 3 times the volume of ethanol solution to the supernatant and let it settle for 12 hours. After centrifugation, take the precipitate and re-dissolve it with water to obtain a crude polysaccharide solution. The Sevage method was used to remove protein, chloroform-n-butanol (5:1), mixed and stirred with the crude polysaccharide solution, and the supernatant was centrifuged. After the end, it was concentrated under reduced pressure at 60°C and freeze-dried for 48 hours to obtain Dendrobium candidum polysaccharide.

[0031] Example 2 Preparation of Dendrobium officinale polysaccharide

[0032] After the Dendrobium officinale is crushed by a grinder, add 20 times the volume of ethanol solution, stir evenly, and soak at room temperature for 12 hours. After centrifugation, discard the supernatant, dry the powder, add 20 times the volume of water, and heat at 100°C for 3 hours. After cooling to room temperature, centrifuge and take the supernatant. Repeat the extraction 3 times, combine the supernatants, and obtain the total polysaccharide extract. Add 4 times the volume of ethanol solution to the supernatant and let it settle for 12 hours. After centrifugation, take the precipitate and dissolve it with water to obtain a crude polysaccharide solution. Use the Sevage method to deproteinize, chloroform-n-butanol (5:1), mix and stir with the crude polysaccharide solution, and centrifuge to obtain the supernatant. After completion, concentrate under reduced pressure at 60°C and freeze-dry for 48 hours to obtain Dendrobium officinale polysaccharide.

[0033] Determination of Molecular Weight of Polysaccharides from Dendrobium officinale

[0034] 1. Preparation of reference solution and test solution

[0035] Take dextran molecular weight standard (D1-D2000) and dissolve it in mobile phase to make 10 mg ml -1 Weigh 10 mg of Dendrobium officinale polysaccharide and dissolve it in mobile phase to make 0.5 mg ml -1 The solution is filtered to obtain.

[0036] 2. Liquid phase conditions

[0037] Chromatographic column: Asahipak GF-7M HQ

[0038] Mobile phase: 0.71% Na2SO4-0.02% NaN 3

[0039] Flow rate: 0.5 mL min -1

[0040] Column temperature: 40°C

[0041] RID detector cell temperature: 40°C

[0042] 3. Determination

[0043] The above solution was taken and tested according to the chromatographic conditions. A standard curve was drawn by GPC software, with the logarithmic value of the relative molecular mass of the reference substance as the ordinate and the retention time of the corresponding chromatographic peak as the abscissa, and regression fitting analysis was performed to calculate the molecular weight distribution of the polysaccharide from Dendrobium officinale.

[0044] 4. Experimental results

[0045] By testing the retention time of dextran standards with different molecular weights and performing curve fitting analysis with GPC software, the regression equation LgMw = -0.000744tR3 + 0.0387tR 2-1.095tR+14.90(R=0.998), standard and fitting curves see Figure 1 and Figure 2 According to the standard curve, the weight average molecular weight (Mw) of the polysaccharide of iron stone is between 716KDa and 102KDa.

[0046] Example 3 Preparation of the composition

[0047] The Dendrobium officinale polysaccharide obtained above and the Brazilian ginseng extract were fully mixed in a mass ratio of 1:4 to prepare a composition.

[0048] Example 4 Preparation of the composition

[0049] The Dendrobium officinale polysaccharide obtained above and the Brazilian ginseng extract were fully mixed in a mass ratio of 2:3 to prepare a composition.

[0050] Animal experiment on the protection of the composition against ethanol-induced acute gastric injury

[0051] 1. Experimental Animals:

[0052] 8-week-old female SPF SD rats, weighing (180-220) g, were purchased from Jiangxi University of Traditional Chinese Medicine.

[0053] 2. Experimental steps:

[0054] After 7 days of adaptive feeding, all rats were randomly divided into 6 groups, with 12 rats in each group, namely blank control group, model group, positive drug group, composition a group, composition b group, polysaccharide group and Brazilian ginseng group.

[0055] Composition a: the mass ratio of Dendrobium officinale polysaccharide and Brazilian ginseng extract is 1:4;

[0056] Composition b: The mass ratio of Dendrobium officinale polysaccharide and Brazilian ginseng extract is 2:3.

[0057] Each group was given the drug once a day for 15 consecutive days. During the drug administration period, the weight, food intake, water intake, and mental state of the rats were recorded. Before the last drug administration, the rats in each group were strictly fasted but not watered for 12 hours. One hour after the last drug administration, except for the blank group, the model group, positive drug group, combination group, polysaccharide group, and Brazilian ginseng group were all given 75% ethanol (6 ml kg -1 ) to construct a gastric mucosal injury model. One hour later, all rats were anesthetized and blood was collected from the abdominal aorta. The stomach of the rats was collected to detect the pathological changes of the stomach and the changes of serum inflammatory factors.

[0058] 3. Experimental results:

[0059] (1) HE staining was used to observe the pathological morphology of rat stomach. Figure 2As shown in the figure, the gastric mucosal tissue structure of rats in the blank control group was intact and clear, the glands were arranged regularly, and there were no abnormal lesions such as tissue defects and cell infiltration. The gastric mucosal tissue glandular structure of rats in the model group was disordered, there was local bleeding, a large amount of epithelial cells fell off, and obvious inflammatory infiltration was also seen. After pretreatment with medication, the gastric mucosa of rats in the positive group, combination group, polysaccharide group and Brazilian ginseng group was well protected. The gastric mucosa was able to maintain structural integrity and regular arrangement of glands, and there were no obvious pathological changes. The symptoms of gastric mucosal injury were alleviated, the shedding of gastric mucosal epithelial cells, gastric mucosal congestion and glandular structure disorder were alleviated, and the infiltration of inflammatory cells was reduced. (2) Effects on the levels of serum TNF-α, IL-6, PG-I, PG-II and GAS in rats, as shown in the figure. Figure 3 As shown, the levels of TNF-α, IL-6, PG-I, PG-II and GAS in the serum of rats in the model group were significantly higher than those in the blank control group, and the difference was statistically significant (P < 0.05, P < 0.01); compared with the model group, the levels of TNF-α, IL-6, PG-I, PG-II and GAS in the serum of rats in the positive group, combination group, polysaccharide group and Brazilian ginseng group were significantly reduced (P < 0.05), and the callback ability of the combination group was better than that of the polysaccharide group and Brazilian ginseng group. Note: Compared with the normal control group, ** P<0.01, *** P<0.01; compared with the model group, # P<0.05, ## P<0.01, ### P<0.05.

[0060] The present invention adopts Dendrobium officinale and Brazilian ginseng, which are both yin-nourishing and stomach-nourishing, to extract polysaccharides and ethanol extracts to prepare a composition. The efficacy of the composition on rats with ethanol-induced gastric mucosal injury model is studied, and the results show that the composition containing Dendrobium officinale polysaccharides and Brazilian ginseng extracts has a good effect of protecting gastric mucosa.

[0061] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. These modifications or replacements will not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A composition having a gastric mucosal protective effect, characterized in that: The composite material is prepared based on dendrobium officinale polysaccharide and Brazilian ginseng extract; the mass ratio of the Brazilian ginseng extract to the dendrobium officinale polysaccharide is 1-2:3-4.

2. A method for preparing a composition having a gastric mucosal protective effect as claimed in claim 1, characterized in that: Preparation of Brazilian ginseng extract: weigh Brazilian ginseng root powder with a mass ratio of 1:1-1:3, add ethanol solution and reflux extract for several times, collect and combine the extract after filtering, and concentrate it into extract liquid in a rotary evaporator to obtain Brazilian ginseng extract; Preparation of Dendrobium officinale polysaccharide: extracting Dendrobium officinale with water and precipitating with alcohol, removing protein with Sevage reagent to obtain a crude polysaccharide solution, rotary evaporating water and organic solvent, and freeze-drying to obtain Dendrobium officinale polysaccharide; The Dendrobium officinale polysaccharide and the Brazilian ginseng extract are fully mixed to obtain the composition.

3. The preparation method according to claim 2, characterized in that: Before the water extraction and alcohol precipitation, the method also includes crushing the Dendrobium officinale, mixing and soaking it with the ethanol solution, and centrifuging it to obtain a first precipitate.

4. The preparation method according to claim 3, characterized in that: The water extraction and alcohol precipitation comprises mixing the first precipitate with water, heating for extraction, centrifuging, taking the supernatant to obtain a total polysaccharide extract; mixing the total polysaccharide extract with an ethanol aqueous solution with a volume fraction of 80%, standing, and taking a second precipitate.

5. The preparation method according to claim 4, characterized in that: The volume ratio of the crushed Dendrobium officinale to the ethanol solution is 1 g: (15-20) mL.

6. The preparation method according to claim 4, characterized in that: The heating treatment conditions are 95-100° C., 2-3 hours, and the extraction times are 2-3 times.

7. The preparation method according to claim 4, characterized in that: The volume ratio of the total polysaccharide extract and the ethanol aqueous solution is 1:(3-4).

8. The preparation method according to claim 4, characterized in that: The molecular weight of the total polysaccharide is 100KDa~800KDa.

9. Use of the composition having gastric mucosa protective effect according to claim 1 in the preparation of a product for nourishing the stomach.

Images