Freeze-dried powder preparation of NK cell extract combined with traditional Chinese medicine and application of freeze-dried powder preparation in treatment of chronic gastritis

Abstract

The invention discloses a freeze-dried powder preparation of an NK cell extract combined with a traditional Chinese medicine and application of the freeze-dried powder preparation in treating chronic gastritis. On the basis of researching the effect of an NK cell extract on treating chronic gastritis, the NK cell extract is wrapped and protected by a nano material, and meanwhile, endothelium corneum gigeriae galli small molecule peptide and a bletilla striata extract are added for compounding to prepare a freeze-dried powder dosage form, so that the effect of treating chronic gastritis is better, the immunity can be improved, and stomach helicobacter pylori infection can be prevented and treated.

Description

Technical Field

[0001] The present invention relates to the technical field of clinical application of cell biology, and in particular to a freeze-dried powder preparation of NK cell extract combined with traditional Chinese medicine and application thereof in treating chronic gastritis. Background Art

[0002] Helicobacter pylori is a gram-negative bacterium that is mainly distributed in the gastric mucosal tissue. 67%-80% of gastric ulcers and 95% of duodenal ulcers are caused by Helicobacter pylori. Common symptoms of patients with chronic gastritis and peptic ulcers are: fullness, discomfort or pain in the upper abdomen after eating, often accompanied by other adverse symptoms such as belching, bloating, acid reflux and loss of appetite. Some patients may also experience recurrent severe abdominal pain, a small amount of upper gastrointestinal bleeding, etc. Clinical studies have shown that Helicobacter pylori infection increases the risk of gastric cancer by 2.7-12 times. If there is no Helicobacter pylori infection, at least 35%-89% of gastric cancer will not occur.

[0003] Helicobacter pylori infects people of different races and regions around the world, and can be said to be the most widespread chronic bacterial infection among adults. The general trend is that the infection rate of Helicobacter pylori increases with age, about 80% in developing countries and about 40% in developed countries, and slightly higher in men than in women.

[0004] At present, most hospitals adopt triple or quadruple therapy of "2 antibiotics + 1 acid inhibitor + 1 prokinetic agent" under the advice of doctors. The course of treatment is 7-14 days. Helicobacter pylori can be turned negative in a short period of time, and stomach discomfort can be relieved. However, due to the large doses of antibiotics used during the treatment, it is extremely destructive to the intestinal flora. Therefore, it didn't take long for the patient to experience symptoms such as acid reflux, stomach bloating, and dull pain. Another examination found that Helicobacter pylori was positive again, and Helicobacter pylori in the body had drug resistance, which increased the difficulty of re-treatment.

[0005] Therefore, treating Helicobacter pylori infection effectively and safely is a difficult problem in today's medical community. Summary of the invention

[0006] The main purpose of the present invention is to provide a freeze-dried powder preparation of NK cell extract combined with traditional Chinese medicine, and its application in treating chronic gastritis. It not only solves the several problems described in the above technical background, but also innovatively develops a product that is more effective than existing gastritis treatment drugs.

[0007] To achieve the above purpose, the present invention, based on the study of the effect of NK cell extracts on the treatment of chronic gastritis, uses nanomaterials to wrap and protect the NK cell extracts, and adds chicken gizzard lining small molecule peptides and bletilla striata extracts to compound them to make a freeze-dried powder dosage form, which not only has a better effect on the treatment of chronic gastritis, but also can prevent gastric infection, enhance immunity, and provide health care and anti-aging.

[0008] NK cell extracts contain NK cell exosomes and NK cell lysates, which have similar biological functions to NK cells. NK cells can not only remove cancerous and diseased cells, but also remove senescent cells, bacteria, viruses, and free radicals. NK cells can enhance human immunity, resist aging, improve sleep, and comprehensively improve metabolic function.

[0009] Chicken gizzard lining has the function of strengthening the stomach and digesting food, which can improve the situation of food stagnation and help relieve the symptoms of indigestion. When chronic superficial gastritis has symptoms of indigestion such as abdominal distension and early satiety, especially when the stomach empties slowly after a meal, resulting in food stagnation, chicken gizzard lining can be eaten.

[0010] Bletilla striata has the effect of astringency and hemostasis, has anti-ulcer effect, and is highly viscous, which can form a gelatinous film in the stomach, which can promote the healing of ulcer surface. In addition, the beneficial ingredients contained in Bletilla striata have antibacterial effects on Staphylococcus, Streptococcus, etc., and can form a protective film, thereby preventing stomach infection.

[0011] The Helicobacter pylori-free gastritis powder (lyophilized powder) prepared with NK cell extract, chicken gizzard lining small molecule peptide and bletilla striata extract as core raw materials has the characteristics of treating Helicobacter pylori infection and chronic atrophic gastritis. Usually, Helicobacter pylori infection can be turned negative and chronic gastritis can be restored to normal after three to five times. It is the most effective means to prevent and treat chronic gastritis and gastric cancer. DETAILED DESCRIPTION

[0012] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0013] In an embodiment of the present invention, a freeze-dried powder preparation of NK cell extract combined with traditional Chinese medicine is provided, wherein the weight proportions of the combined traditional Chinese medicine are: 0.8-1.3 parts of NK cell extract, 0.8-1.3 parts of chicken gizzard lining small molecule peptide, and 0.8-1.3 parts of Bletilla striata extract. Among them:

[0014] NK cell extracts contain NK cell exosomes and NK cell lysates, which have similar biological functions to NK cells. NK cells can not only remove cancerous and diseased cells, but also remove senescent cells, bacteria, viruses, and free radicals. NK cells can enhance human immunity, resist aging, improve sleep, and comprehensively improve metabolic function.

[0015] NK cells, also known as natural killer cells (NK cells), are important immune cells in the body. NK cells are large granular lymphocytes in morphology and are derived from the bone marrow. They are the third largest type of lymphocytes besides T cells and B cells, accounting for about 15% of all immune cells (white blood cell count) in the blood. They are core cells of the innate immune system and are mainly distributed in peripheral blood, liver and spleen. The main characteristics of NK cells in the human body are CD3-CD56+ lymphocyte populations, of which the blood is mainly CD16+CD56dim subtype (based on the difference in the expression density of CD56 molecules on cells, NK cells are divided into two subgroups, CD56dim and CD56bright; CD56dim accounts for more than 90% of NK cells, mainly for cytotoxic effects, expressing moderately affinity IL-2 receptors (IL-2R), and has stronger killing activity; CD56bright can produce a large number of cytokines, mainly for immunomodulatory effects, and highly expresses IL-2R).

[0016] NK cells are the main "fighters" responsible for killing abnormal cells such as aging, virus-infected, and tumor cells in the body. NK cells are different from more than 150 other white blood cells in the human body. They do not need to receive special instructions from the immune system, nor do they need the cooperation of other cells. They can identify and attack foreign cells, cancer cells, and viruses on their own. Circulating NK cells are usually dormant. Once activated, they will infiltrate into tissues, secrete perforin and tumor necrosis factor, and attack tumor cells and virus-infected cells. NK cells are the first line of defense for the human body against cancer cells and viral infections. They can directly kill tumor cells non-specifically. This natural killing activity does not require antigen sensitization or antibody participation, and is not restricted by MHC. In addition to its powerful killing function, it also has a strong immune regulation function, interacting with a variety of other immune cells in the body to regulate the body's immune status and immune function.

[0017] NK cells can release perforin and granzyme. Perforin perforates the surface of target cells, allowing granzyme b to enter target cells and induce apoptosis of target cells. At the same time, they secrete a large number of cytokines, such as IFN-γ, TNF-ɑ, GM-CSF, IL-3, M-CSF, etc., which directly act on target cells or further activate other types of immune cells to attack target cells. They can also express proteins that can induce cell apoptosis (FAS-L) and tumor necrosis factor-related apoptosis-inducing ligand (Trail), causing target cells to enter a state of programmed apoptosis.

[0018] Chicken gizzard lining extract has pharmacological effects such as regulating digestive system function, blood system function, improving blood rheology, and inhibiting the growth of myoma.

[0019] 1) Effects on human gastric function

[0020] Healthy people take 5g of chicken gizzard powder, and after 45-60 minutes, the gastric juice secretion increases by 30-37% compared with the control value, and returns to normal within 2 hours. The acidity of gastric juice also increases significantly. Free acid or total acidity generally begins to rise 1 hour after taking chicken gizzard, reaches the highest value in 1-2 hours, and then gradually decreases, and returns to normal after 3 hours. Among them, the highest value of free acid is 0.19-0.24%, which is 32-113% higher than the control value; the highest value of total acidity is 0.2-0.32, which is 25-75% higher than the normal value. Although the increase in digestive power is slow, it lasts for a long time. Gastric motility can be significantly enhanced, which is manifested in prolonged gastric motility and enhanced peristaltic waves, so the gastric emptying rate is accelerated. Chicken gizzard itself contains only trace amounts of pepsin and amylase. After taking the medicine, it can increase the secretion of gastric juice and enhance gastric motility. It is believed that this may be caused by the excitement of the nerves and muscles of the stomach wall through humoral factors after digestion and absorption of chicken gizzard, and it is also believed that gastric hormones promote gastric secretion function.

[0021] 2) Regulate the secretion of digestive juice

[0022] There are also many studies on the effect of chicken gizzard lining on gastric secretion. Its main mechanism is to regulate the secretion of digestive juice by significantly increasing the activity of gastric pepsin. Taking gastric juice secretion and pepsin excretion as reference indicators, the effects of raw products and different processed products on the secretion function of rat digestive juice were investigated. The results showed that each drug group could significantly increase the above two indicators; and compared with the raw product, each processed product could regulate the secretion of digestive juice to varying degrees.

[0023] 3) Gastrointestinal health

[0024] Gastric mucosal damage is a pathological manifestation of patients with gastric diseases. Experimental studies have confirmed that the commonly used chicken gizzard lining can activate factors related to the protection of gastric mucosa, improve the discomfort caused by gastric mucosal damage, adjust gastrointestinal function, and have fewer adverse reactions. The charcoal powder small intestine propulsion method was used to study the effect of gastrin, an active ingredient in chicken gizzard lining extract, on the optimization of the intestinal health function of mice. It was found that different doses of the extract had different degrees of optimization of the intestinal health function of mice, indicating that the high-dose group can significantly improve gastrointestinal function and enhance the propulsion movement of the small intestine, while there was no significant change in the low and medium dose groups.

[0025] 4) Anti-cancer effect

[0026] In vitro tests have shown that chicken's gizzard lining has the effect of inhibiting tumor cells. Oral administration of chicken's gizzard lining increases gastric juice secretion, acidity and digestive capacity, strengthens gastric motility, accelerates emptying rate, and has a secretion-inducing effect that is even stronger than meat powder. The pathway of action is that it enters the blood after digestion and absorption, and excites the neuromuscular apparatus of the stomach wall through humoral factors.

[0027] Bletilla striata has the effect of astringency and hemostasis, has anti-ulcer effect, and is highly viscous, which can form a gelatinous film in the stomach, which can promote the healing of ulcer surface. In addition, the beneficial ingredients contained in Bletilla striata have antibacterial effects on Staphylococcus, Streptococcus, etc., and can form a protective film, thereby preventing stomach infection.

[0028] Studies have shown that Bletilla polysaccharide can reduce the malondialdehyde content in rats with stress ulcers. The principle of its protective effect on gastric mucosa is to increase the activity of superoxide dismutase. Studies have found that Bletilla polysaccharide has a significant therapeutic effect on acetic acid gastric ulcers in rats. Its mechanism is related to enhancing the antioxidant capacity of gastric mucosa, promoting the proliferation of local gastric mucosal epithelial cells in gastric ulcers, inhibiting the generation of free radicals, and strengthening the repair of damaged tissues. In the experiment on rat stress gastric ulcers, it was found that Bletilla polysaccharide has a significant anti-stress gastric ulcer effect. This effect is related to the fact that Bletilla polysaccharide enhances the barrier and defense function of gastric mucosa, reduces the damage of gastric mucosa to attack factors, and enhances the ability to scavenge free radicals.

[0029] Bletilla striata has bactericidal and anti-inflammatory effects. The ethanol extract of Bletilla striata diluted by the plate dilution method has an inhibitory effect on Bacillus subtilis at 1:20 and on Staphylococcus aureus and Mycobacterium tuberculosis at 1:100. The decoction of Bletilla striata at 1g / mL can inhibit Streptococcus mutans. Dihydrophenanthrene and biphenyl compounds isolated from Bletilla striata tubers have an inhibitory effect on Staphylococcus aureus, Bacillus cirrhosa QM248 and Bacillus subtilis. Studies have found that the structure of pathogenic microorganisms is related to the antibacterial activity of Bletilla striata, and its antibacterial activity against hydroxybenzyl compounds is enhanced, while its antibacterial activity against methoxy compounds is weakened.

[0030] The Helicobacter pylori-free gastritis powder (lyophilized powder) prepared with NK cell extract, chicken gizzard lining small molecule peptide and Bletilla striata extract as core raw materials is effective in treating Helicobacter infection and chronic atrophic gastritis. Usually, three to five times can turn the Helicobacter pylori infection to negative and chronic gastritis back to normal. It is the most effective means to prevent and treat chronic gastritis and gastric cancer.

[0031] The present invention is specially compared with Xiangsha Yangwei Pills in the efficacy of treating spleen and stomach deficiency and cold syndrome.

[0032] There were 50 cases in this group, which were randomly divided into a treatment group of 25 cases and a control group of 25 cases. In order to make the two groups comparable, we compared the age, gender, course of disease, and condition of the two groups, as shown in Tables 1-4.

[0033] Table 1 Comparison of age distribution between the two groups (years)

[0034]

[0035] As can be seen from Table 1, there is no significant difference in the age distribution between the two groups, P>0.05, which means that the age distribution of the two groups is comparable.

[0036] Table 2 Comparison of gender distribution between the two groups

[0037]

[0038] As can be seen from Table 2, there was no significant difference in gender distribution between the two groups after statistical processing, P>0.05.

[0039] Table 3 Comparison of disease course distribution between the two groups (months)

[0040]

[0041] As can be seen from Table 3, there was no significant difference in the distribution of disease course between the two groups after statistical analysis, P>0.05.

[0042] Table 4 Comparison of disease severity between the two groups before treatment

[0043]

[0044] As can be seen from Table 4, the comparison of the disease severity between the two groups before treatment, after statistical processing, P>0.05, indicating that the disease severity of the two groups is balanced and comparable.

[0045] 1. Selection of subjects

[0046] 1.1. TCM diagnostic criteria:

[0047] (1) Epigastric pain and gastrointestinal symptoms;

[0048] (2) recurrent attacks;

[0049] (3) Symptoms occur or worsen after exertion or exposure to cold.

[0050] 1.2. Chinese medicine dialectics:

[0051] Dull pain in the stomach, continuous, prefers warmth and heat, worse on an empty stomach, relieved by food, vomiting of clear water, fatigue, poor appetite, cold hands and feet, loose and thin stools, pale tongue with white coating, and weak or slow pulse.

[0052] 1.3. For details on the severity of TCM symptoms, see Table 5.

[0053] Table 5 TCM symptom severity classification table

[0054]

[0055] 2. Test methods

[0056] 2.1 Treatment methods

[0057] (1) The treatment group of 25 cases orally took the lyophilized powder of the Chinese medicine composition of the present invention, once a day, 1 bottle each time, for 3 days. (2) The control group of 25 cases selected Xiangsha Yangwei Pills, 2 bags each time, 3 times a day, for 3 days.

[0058] 2.2 Observation indicators

[0059] (1) The degree, nature and duration of dull pain in the stomach.

[0060] (2) Symptoms and signs associated with dull pain in the stomach.

[0061] 2.3 Test results

[0062] 2.4. Criteria for evaluating efficacy

[0063] (1) Recovery: The main symptoms disappear and do not recur within six months to one year.

[0064] (2) Significantly effective: Most of the main symptoms disappear and do not recur within 6 months to 1 year.

[0065] (3) Effective: The main symptoms basically disappear, and there will be recurrence within half a year to a year, but the severity will be reduced and the duration will be shortened. (4) Ineffective: The main symptoms remain unchanged.

[0066] 2.5. Experimental results and efficacy analysis

[0067] The efficacy of 50 patients observed in this study is shown in Table 6.

[0068] Table 6 Comparison of the efficacy between the two groups

[0069]

[0070] The results showed that the total effective rate of the treatment group was 100%, and the total effective rate of the control group was 76.00%, with a P>0.05 between the two groups. However, there was a significant difference in the cure rate between the treatment group and the control group, P<0.01.

[0071] The test results show that the efficacy of the Chinese medicine combination of the present invention and Xiangsha Yangwei Pills in treating spleen and stomach deficiency and cold syndrome is significantly different, and the cure rate of the Chinese medicine combination of the present invention is significantly higher than that of the control group. This provides an objective test basis for guiding clinical medication.

[0072] 3. Toxicity test

[0073] Take 20 mice, half of which are male and half are female, and administer the drug of the present invention to the mice three times a day at a concentration of 30.6% and a volume of 40 ml / kg. After administration, the mice are observed for seven consecutive days. None of the animals died, and the appearance, behavior, mental state, appetite, urine and stool, color, fur, skin color, and breathing were normal. There was no abnormal secretion in the nose, eyes, and mouth, and the body weight increased from 18.2±0.5g to 21.7±1.0g (X±SD). This result shows that the Chinese medicine combination of the present invention administered to mice by oral administration did not cause acute toxic reactions in the animals.

[0074] Example 1

[0075] Liu XX, male, 57 years old, self-reported: he has a history of gastric disease for more than 20 years. The results of the hospital test at the end of 2014 were: Helicobacter pylori positive, and the conclusion of gastroscopy was severe atrophy, severe intestinal metaplasia and mild atypical hyperplasia. He had been treated with triple and quadruple therapy three times in a row since 2013, and Helicobacter pylori was still positive. On January 21, 2015, he began to use the freeze-dried powder preparation of the NK cell extract combined with traditional Chinese medicine of the present invention for treatment. From January 21 to 23, he took the freeze-dried powder preparation of the NK cell extract combined with traditional Chinese medicine of the present invention orally every day for 3 consecutive days. On the second day after taking the medicine, he felt that his symptoms such as stomach pain, abdominal distension, acid reflux and loss of appetite were improved. As of November 2018, he had multiple physical examinations for Helicobacter pylori and they were all negative. In November 2018, the hospital test results were: Helicobacter pylori negative, and the conclusion of gastroscopy was chronic superficial gastritis.

[0076] Example 2

[0077] Zhao XX, male, 36 years old, self-reported: in the past 3-5 years, he occasionally had symptoms such as upper abdominal burning pain, abdominal distension, acid reflux and irregular bloody stools. The results of the hospital test in 2015: diagnosed with gastritis and duodenal ulcer, Helicobacter pylori positive, etc. After more than a year of conventional treatment, the condition recurred and the Helicobacter pylori indicators did not improve. In January 2017, he took oral NK cell extract combined with freeze-dried powder preparations of traditional Chinese medicine for treatment. After treatment, he felt that the symptoms of upper abdominal burning pain, abdominal distension, acid reflux, blood in the stool were significantly improved. On the 30th day after treatment, the Helicobacter pylori test was still positive, and on the 60th day, the hospital test for Helicobacter pylori turned negative. As of October 2018, the latest hospital test result: Helicobacter pylori negative.

[0078] Example 3

[0079] Guan XX, female, 32 years old, self-reported: stomach pain, abnormal stool, severe oral odor and other symptoms in the past six months. The results of the hospital test in 2016: diagnosed with atrophic gastritis, Helicobacter pylori positive, anemia, etc. After two months of regular Chinese and Western medicine treatment, the condition recurred, and the symptoms and indicators did not improve. In March 2017, she began to use NK cell extract combined with Chinese medicine freeze-dried powder preparations for treatment. After treatment, the symptoms of stomach pain, abnormal stool, severe oral odor, etc. were significantly improved. On the 30th day after treatment, the Helicobacter pylori test turned negative, and all clinical symptoms disappeared.

[0080] The above description is only a preferred embodiment of the present invention, and does not limit the patent scope of the present invention. All equivalent structural changes made by using the contents of the present invention specification under the inventive concept of the present invention, or directly / indirectly applied in other related technical fields are included in the patent protection scope of the present invention.

Claims

1. A freeze-dried powder preparation of NK cell extract combined with traditional Chinese medicine, characterized in that: The weight proportions of the combined traditional Chinese medicine are as follows: 0.8-1.3 parts of NK cell extract, 0.8-1.3 parts of chicken gizzard lining small molecule peptide, and 0.8-1.3 parts of bletilla striata extract.

2. A freeze-dried powder preparation of NK cell extract combined with traditional Chinese medicine according to claim 1, characterized in that: The weight proportions of the combined traditional Chinese medicine are as follows: 0.9-1.2 parts of NK cell extract, 0.9-1.2 parts of chicken gizzard lining small molecule peptide, and 0.9-1.2 parts of bletilla striata extract.

3. The freeze-dried powder preparation of NK cell extract combined with traditional Chinese medicine according to claim 1, characterized in that: The weight proportions of the combined traditional Chinese medicine are as follows: 1 part of NK cell extract, 1 part of chicken gizzard lining small molecule peptide, and 1 part of bletilla striata extract.

4. The freeze-dried powder preparation of NK cell extract combined with traditional Chinese medicine according to claim 1, characterized in that: The weight proportions of the combined traditional Chinese medicine are as follows: 0.8 parts of NK cell extract, 1 part of chicken gizzard lining small molecule peptide, and 1.2 parts of bletilla striata extract.

5. The NK cell extract combined with traditional Chinese medicine according to claim 1, characterized in that: The weight proportions of the combined traditional Chinese medicine are as follows: 1.1 parts of NK cell extract, 0.9 parts of chicken gizzard lining small molecule peptide, and 1 part of bletilla striata extract.

6. Application of a freeze-dried powder preparation of NK cell extract combined with traditional Chinese medicine in the treatment of chronic gastritis, characterized in that: A freeze-dried powder preparation of the NK cell extract combined with traditional Chinese medicine as described in any one of claims 1 to 5.

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