Medicinal and edible composition with function of treating helicobacter pylori related gastritis as well as preparation method and application of medicinal and edible composition

Abstract

The invention discloses a medicinal and edible composition with a function of treating helicobacter pylori related gastritis as well as a preparation method and application thereof, and the medicinal and edible composition comprises the following components in parts by weight: 6-12 parts of ginseng, 12-24 parts of poria cocos, 9-18 parts of raw liquorice, 9-18 parts of pogostemon cablin, 9-18 parts of pericarpium citri reticulatae and 15-30 parts of dandelion. The prescription of the helicobacter pylori-related stomach-protecting paste follows the traditional Chinese medicine theory, has the effects of replenishing qi to invigorate the spleen and eliminating dampness and heat, and is used for carrying out auxiliary conditioning on the pathogenesis of helicobacter pylori-related gastritis spleen and stomach qi deficiency and damp-heat accumulation; the product not only has certain efficacy, but also has color, aroma, taste and shape as food, and is better in taste; the preparation process is simple, is beneficial to industrial production and has wide market prospects.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of health food, in particular to a kind of homologous composition of medicine and food with the function of treating Helicobacter pylori associated gastritis and its preparation method and application. BACKGROUND

[0002] Helicobacter pylori (H.pylori) is a gram-negative, microaerophilic bacteria, which colonizes between gastric mucus layer and antral mucosal epithelial cells. It can destroy the gastric mucus-bicarbonate barrier, induce immune response, promote the release of inflammatory mediators, induce gastric mucosal injury, and cause DNA damage, leading to genomic instability and gastric cancer. The inflammation of gastric mucosa after H.pylori infection is called Helicobacter pylori associated gastritis (HPAG), which is characterized by different degrees of stomach pain, abdominal distension, acid vomiting and other symptoms. Guidelines recommend a triple therapy regimen (i.e. proton pump inhibitor combined with clarithromycin, amoxicillin or metronidazole) or a quadruple therapy regimen containing bismuth. Due to the widespread and long-term use of antibiotics, there are many adverse reactions, such as increasing drug resistance, digestive dysfunction, and increasing recurrence rate. Therefore, it is urgent to develop new safe and low-toxic health products. Studies have shown that traditional Chinese medicine has significant advantages in improving H.pylori eradication rate and can improve gastric mucosal damage caused by H.pylori infection. Traditional Chinese medicine has long held the belief that "medicines and food are homologous". Both medicines and food come from nature, and the concept of homology between medicine and food can be traced back to ancient times. Medicinal herbs in the homology of medicine and food are increasingly popular among the public due to their multiple effects, and are a trend in functional foods.

[0003] Traditional Chinese medicine decoction is commonly used to treat Helicobacter pylori associated gastritis. However, traditional decoction has poor taste and is difficult to adhere to long-term use, resulting in low patient compliance. In recent years, homologous products of medicine and food have been widely promoted in the market with the support of relevant policies and the development of related universities and hospitals. Functional foods of medicine and food mainly use medicinal herbs that can be used for both medicine and food. Under the guidance of theories of traditional Chinese medicine, nutrition, pharmacy and other disciplines, they are made from cereals, fruits and vegetables and other food materials through process optimization to have specific health care functions. They not only have the function of preventing and protecting health, but also meet the food properties of color, aroma and taste.

[0004] Sijunzi Decoction is from Taiping Huimin Heji Ju Fang, and it is recorded in the book that "Rongweiqi is weak, Zangfu is weak …… all not thinking about food and drink, intestinal rumbling, diarrhea, vomiting, too suitable for taking it". It is used for treating spleen-stomach qi deficiency syndrome. The symptoms include sallow complexion, shortness of breath, lack of strength, less food and loose stool, pale tongue, white fur and weak pulse. Sijunzi Decoction has the remarkable effect of benefiting qi and invigorating the spleen, and is often used as a basic prescription for the treatment of diseases related to spleen deficiency, such as gastric ulcer, gastritis, duodenal gastritis, ulcerative colitis, etc. Spleen and stomach are in the middle of the body, and they are mutually beneficial and complementary. The ascending and descending of qi, the transportation of water and food, and the absorption of nutrients are all related to each other. Therefore, spleen and stomach are also called the basis of acquired constitution. The Su Wen-Jing Mai Bian Lun Chapter states that "spleen and stomach are the officials of the granary, and five flavors come from them", which discusses the function of spleen and stomach. The Su Wen-Jing Mai Bian Lun Chapter records that "food enters the stomach and spreads essence to the liver …… drink enters the stomach and spreads essence …… spleen qi spreads essence and returns to the lung". These discussions point out that both spleen and stomach have the function of transporting and transforming water and essence.

[0005] The Shanhaijing states that "taste is good and all slippery is paste", which points out that paste is the essence of the material and is also nourishing. There are eight kinds of traditional Chinese medicine dosage forms, including pill, powder, paste, pill, decoction, wine, dew and tablet. "Paste" refers to a thick paste, which contains the meaning of nourishment, and can be divided into external use and internal use. External paste is generally called "plaster"; paste prescription is also called "decoction paste" and "paste", which is one of the oldest dosage forms. Internal paste is prepared under the guidance of traditional Chinese medicine theory, in accordance with the traditional prescription principles, using medicinal materials with the comprehensive effects of nourishing and treatment and prevention. On the basis of compound decoction, the prescription is formulated according to the different constitutions and clinical manifestations of patients. After cleaning, soaking, decocting, evaporating and concentrating of traditional Chinese medicine decoction pieces, or adding refined honey, refined sugar and other auxiliary materials, a semi-liquid preparation is prepared. Therefore, a kind of food-medicine combination composition is needed for the auxiliary treatment of Helicobacter pylori related gastritis. SUMMARY

[0006] The purpose of the present application is to solve the problems in the prior art, and to provide a food-medicine combination composition with the function of treating Helicobacter pylori related gastritis, and its preparation method and application. The composition has the functions of benefiting qi, invigorating the spleen and clearing and draining damp heat, and is suitable for the auxiliary treatment of Helicobacter pylori related gastritis.

[0007] In order to solve the above technical problems, the technical scheme of the present application is as follows:

[0008] A food-medicine combination composition with the function of treating Helicobacter pylori related gastritis, the food-medicine combination composition comprises, by weight, 6-12 parts of ginseng, 12-24 parts of poria, 9-18 parts of raw licorice, 9-18 parts of patchouli, 9-18 parts of dried tangerine or orange peel, and 15-30 parts of dandelion.

[0009] The Qingyou Hugan Paste takes ginseng as the monarch drug, tangerine peel and poria as the minister drugs, pogostemonis herba and dandelion as the auxiliary drugs, and raw licorice as the ministerial drug. Ginseng can nourish fluid and tonify spleen qi, poria can slightly penetrate and drain dampness, tangerine peel can dry dampness and invigorate the spleen, pogostemonis herba can resolve dampness and harmonize the stomach, dandelion can resolve toxicity and dry dampness, and raw licorice can regulate qi and relieve pain, so that the whole prescription can invigorate the stomach and nourish the spleen, resolve dampness and resolve toxicity, and is used for treating the spleen-stomach qi deficiency syndrome and the damp-heat accumulation syndrome of HPAG.

[0010] In a preferred embodiment of the present application, the medicinal and edible composition further comprises honey 60-120 parts by weight.

[0011] Preferably, the medicinal and edible composition comprises ginseng 6-10 parts, poria 12-16 parts, raw licorice 9-12 parts, pogostemonis herba 9-12 parts, tangerine peel 9-12 parts, dandelion 15-20 parts, and honey 60-80 parts. Further preferably, the medicinal and edible composition comprises ginseng 6-8 parts, poria 12-14 parts, raw licorice 9-10 parts, pogostemonis herba 9-10 parts, tangerine peel 9-10 parts, dandelion 15-16 parts, and honey 60-70 parts.

[0012] Preferably, the Qingyou Hugan Paste comprises the following raw materials by weight:

[0013] Ginseng 6 parts, poria 12 parts, raw licorice 9 parts, pogostemonis herba 9 parts, tangerine peel 9 parts, dandelion 15 parts, and honey 60 parts.

[0014] The present application further discloses a preparation method of the medicinal and edible composition with the function of treating H. pylori associated gastritis.

[0015] S1, soaking ginseng, poria, raw licorice, pogostemonis herba, tangerine peel and dandelion;

[0016] S2, mixing ginseng, poria, raw licorice, pogostemonis herba, tangerine peel and dandelion after soaking in S1 and then performing water extraction;

[0017] S3, mixing the extraction liquid obtained through water extraction in S2 and then concentrating to obtain a clear paste;

[0018] S4, heating and concentrating honey and then cooling to obtain refined honey;

[0019] S5, adding the refined honey in S4 to the clear paste prepared in S3 and then continuing to concentrate.

[0020] In a preferred embodiment of the present application, the soaking time in step S1 is at least 120 minutes. After the medicinal materials are soaked for 120 minutes, the water absorption of the medicinal materials has basically reached a saturated state.

[0021] In a preferred embodiment of the present application, in step S2, the decoction ratio is 1: (8-15), the decoction time is 1-3 hours, and the decoction is performed 2-4 times.

[0022] Preferably, in step S2, the decoction ratio is 1: (9-11), the decoction time is 1-2 hours, and the decoction is performed 3-4 times. Under this condition, the dry extract yield is obviously higher than that under other conditions.

[0023] In a preferred embodiment of the present application, in step S3, the relative density of the clear extract is 1.1-1.3.

[0024] In a preferred embodiment of the present application, in step S4, the honey is heated in a pot, the temperature reaches 116-118℃, the water content is 14-16%, and the relative density is 1.2-1.4 (20℃). The refined honey can remove impurities, kill microorganisms, reduce the water content in the extract, destroy enzymes, increase adhesion, and prolong the storage period of the extract.

[0025] In S2, the water extraction is performed at least three times, and in S3, the extraction liquids obtained by multiple water extractions are mixed.

[0026] Further, in step S3, the concentration is performed under the conditions of 45-55℃, 60-80R / min, and 35-45h Pa.

[0027] In S5, the mass ratio of the clear extract and the refined honey is (1-2) : (1-2).

[0028] In S5, the heating is stopped after the "stringing" state appears.

[0029] S6, the extract obtained in S5 is placed in a ceramic bowl for high-temperature sterilization treatment, the extract is cooled for 12-16 hours, and then is filled into a high-temperature sterilized glass bottle and stored in a 4℃ refrigerator.

[0030] The present application also discloses a kind of medicinal and edible composition or medicinal and edible composition prepared by the preparation method described in the present application is applied to prepare medicinal and edible preparation for treating Helicobacter pylori related gastritis.

[0031] In a preferred embodiment of the present application, the medicinal and edible composition is applied to prepare a preparation for reversing the decrease of local pro-inflammatory factor IL-18 and / or TNF-α and / or the increase of IL-10 in intestinal inflammation.

[0032] In a preferred embodiment of the present application, the medicinal and edible composition is applied to prepare a preparation for reversing the increase of local oxidative stress factor SOD and / or the decrease of MDA in intestinal inflammation.

[0033] Qingyou Hugan Gao is modified from the classic Sijunzi Decoction, which is recorded in the Treatise on Cold Pathogenic and Sudden Febrile Diseases. The Sijunzi Decoction is from the Imperial Medical Bureau Prescriptions for Popular and Domestic Relief, which records that “the vital energy of the lung and heart is weak, and the viscera are weak and feeble …… the patient has no desire to eat and drink, has loose bowels, and vomits. It is suitable to take the Sijunzi Decoction”. The Sijunzi Decoction is used to treat deficiency of the spleen and stomach. The patient has a pale face, shortness of breath, lack of energy, less food and loose bowels, pale tongue, white coating, and weak pulse. The Sijunzi Decoction has a significant effect of tonifying the spleen and stomach and is often used as a basic prescription for the treatment of diseases related to deficiency of the spleen and stomach, such as gastric ulcer, gastritis, duodenal gastritis, and ulcerative colitis.

[0034] The spleen and stomach are located in the middle of the body, and the yin and yang, dryness and dampness, and the ascending and descending of the qi are mutually related. The water and food are absorbed, digested, and absorbed by the spleen and stomach, and the refined substances are transported to the whole body. Therefore, the spleen and stomach are also called the foundation of acquired constitution. The Treatise on the Channels and Vessels states that “the spleen and stomach are the officials of the granary, and the five flavors come from them”. The Treatise on the Channels and Vessels records that “food enters the stomach and is refined in the liver …… drink enters the stomach and is refined in the liver …… the spleen qi disperses the refined substances and returns to the lung”. These statements point out that the spleen and stomach have the function of refining the refined substances of water and food. The Treatise on the Channels and Vessels points out that “sweetness nourishes the spleen” and that the spleen deficiency should be treated with sweetness, which points out the basic treatment plan for the spleen deficiency.

[0035] The modified prescription of the paste is composed by the doctor according to the constitution and symptoms of the patient, which fully embodies the characteristics of syndrome differentiation and treatment, and the treatment according to the person, time, and place. The paste is generally added with sugar or honey to adjust the taste, and the taste is better than that of the general decoction. After extraction and concentration, the volume of the medicine is reduced, the concentration of the medicine is improved, and the medicine is easy to store. The traditional purpose of the paste is to adjust and relieve the symptoms, and the paste can be used for a long time, especially for the patients with chronic diseases, such as the people with weak spleen and stomach, to achieve the purpose of strengthening the body resistance and expelling the evil.

[0036] The traditional Chinese medicine prescription of the Qingyou Hugan Gao is modified from the Sijunzi Decoction. The invention is a food with the same origin as medicine, and the non-food-medicine-attributed drug atractylodes is removed. Considering that the gastric antrum helicobacter related gastritis has symptoms such as abdominal pain caused by qi stagnation, the invention replaces the prepared licorice with raw licorice, and the whole prescription has the same effect.

[0037] The prescription used in the invention is modified from the Sijunzi Decoction and is composed of six medicines, including ginseng, poria, licorice, dried tangerine or orange peel, patchouli, and dandelion, which are all medicines with the same origin as food. The ginseng supplements the qi of five viscera, the poria invigorates the spleen and removes dampness, the licorice supplements the middle jiao and detoxifies and moves qi, the dried tangerine or orange peel dries dampness and moves qi, the patchouli resolves dampness and harmonizes the stomach, and the dandelion clears heat and resolves toxins. The whole prescription has the functions of invigorating the spleen and stomach, resolving dampness and detoxifying.

[0038] Ginseng is the dried root and rhizome of Panax ginseng C. A. Mey. of the Araliaceae family. It has a sweet and slightly bitter taste, and is warm in nature. It is associated with the spleen, lung, heart and kidney meridians. It has the functions of tonifying vital energy and invigorating the spleen. It is used to treat deficiency of vital energy, poor appetite, fatigue, and other deficiency syndromes. It has a long history of use as a medicine, and was first recorded in the Shennong Bencao Jing. It is said that long-term use can prolong life. Studies have found that ginseng polysaccharides have an anti-H. pylori adhesion effect. Another study found that red ginseng extract inhibited the release of inflammatory mediators (IL-1β, iNOS) in the gastric mucosa of infected Mongolian gerbils, and reduced lipid peroxide (LPO) and myeloperoxidase (MPO) levels.

[0039] Gancao is derived from the dried roots and rhizomes of Glycyrrhiza uralensis Fisch., Glycyrrhiza flata Bat., and Glycyrrhiza glabra L. of the Leguminosae family. It is neutral in nature and sweet in taste. It is associated with the heart, lung, and spleen-stomach meridians. It has the functions of tonifying the spleen and invigorating qi, clearing heat and resolving toxins, and relieving acute and chronic cough. It is used for spleen-stomach weakness, fatigue, and abdominal pain. The main active components of gancao are total flavonoids, polysaccharides, and triterpenoid saponins. The pharmacological effects mainly include anti-inflammatory, anti-tumor, antibacterial, antioxidant, and immune regulation. Animal experiments have found that glycyrrhizic acid has a protective effect on the gastric mucosa of H. ploryi-infected Mongolian gerbils. Glycyrrhizic acid can reduce the expression levels of TNF-α, IL-1β, COX-2, and iNOS mRNA in the gastric mucosa of the model group, and reduce inflammation.

[0040] Fuling is the dried sclerotium of Poria cocos (Schw.) Wolf of the Polyporaceae family. It has a sweet and bland taste, and is neutral in nature. It has the functions of promoting water excretion and dampness, invigorating the spleen and stomach, and calming the heart and soothing the spirit. Modern research has found that it contains triterpenes, polysaccharides, and other chemical components, and has anti-inflammatory, liver-protective, antioxidant, and anti-aging effects. Animal experiments have found that the extract of Fuling can promote the proliferation of human gastric mucosa epithelial cells damaged by alcohol. Fuling total triterpenes and Fuling acidic polysaccharides can enhance the repair capacity of the gastric mucosa of spleen-deficient rats by increasing the expression of epidermal growth factor (EGF) and epidermal growth factor receptor (EGFR) in the serum.

[0041] Pericarpium Citri Reticulatae is the dried mature pericarp of Citrus reticulata Blanco and its cultivated varieties. Studies have shown that Pericarpium Citri Reticulatae mainly contains flavonoids, alkaloids, volatile oils and other active ingredients, among which the main flavonoids are nobiletin and citropten. Pericarpium Citri Reticulatae has a unique aroma and is widely used in food and medicine. Studies have found that nobiletin can effectively reduce the levels of IL-6, IL-1β and TNF-α in mouse stomach tissue, achieving the effect of repairing ethanol-induced ulcer mucosa damage.

[0042] Taraxacum mongolicum Hand.-Mazz., Taraxacum borealisinense Kitam. or several species of the same genus, is a dry whole plant with bitter and sweet taste, cold nature, and liver and stomach meridians. It has the effects of clearing heat and resolving toxins, resolving and removing stasis, and diuresis and blood circulation. It is mainly used in the treatment of furuncle, sore, sore throat, mastitis, lung abscess, intestinal abscess, and other diseases. Its active ingredients include flavonoids, phenolic acids, poly-alpha sugars, terpenes, sterols, etc. Its pharmacological effects include anti-inflammatory, antioxidant, and anti-tumor effects.

[0043] Pogostemon cablin (Blanco) Benth. is a dry aboveground part of the Lamiaceae plant Pogostemon cablin (Blanco) Benth. It has a bitter and sweet taste, a mild warm nature, and a spleen, stomach, and lung meridian. It has the effects of aromatic and turbidity, and stomach and vomiting, and can dispel heat and relieve the exterior. In ancient books, Pogostemon cablin (Blanco) Benth. is mainly used as Pogostemon cablin (Blanco) Benth. It originated in Southeast Asian countries and was initially widely used as a spice. Later, it was gradually used as a good medicine for dispelling heat and relieving dampness. Since the Song Dynasty, it has been introduced to Guangdong and Guangxi regions of China and is one of the famous "Ten South Medicines". The main active ingredient of Pogostemon cablin (Blanco) Benth. is volatile oil, and pogostol is one of the main components of volatile oil. Pogostol has anti-inflammatory, antibacterial, and gastrointestinal protection effects. Its treatment indications are similar to the symptoms of HPA, such as hiccup, halitosis, and fullness in the epigastric region.

[0044] Honey has the effects of seasoning and tonifying. Refined honey can remove impurities, kill microorganisms, reduce the water content in the paste, destroy enzymes, increase adhesion, and prolong the shelf life of the paste.

[0045] Jing Zhangxugui said: "Wet-heat evil, although the beginning of external, eventually return to the spleen and stomach also". "Ling Sou: abscesses" "big heat, heat, then the meat rot". Plus wet evil heavy turbidity sticky, wet heat mutual cementation of the stomach and intestines, then easy to breed outside evil. As "medical" said: "wet heat, worms, like a ditch dirty accumulated for a long time not flow, then the insects in it". If the spleen and stomach weakness, plus wet heat invasion, the disease is easy to repeat infection difficult to eradicate. Studies have shown that the highest infection rate of chronic gastritis syndrome of spleen and stomach dampness, wet evil and H. pylori gas together, the two interact, so the disease is lingering difficult to cure. H. pylori listed in the category of TCM "evil", and "wet heat", "toxic evil" most closely related.

[0046] Qingyouhugan paste with four gentlemen decoction as the basis of the prescription, with food and medicine similar traditional Chinese medicine, with the effect of benefiting qi and invigorating the spleen, clearing and removing damp-heat, aiming at the pathogenesis of H. pylori related gastritis with spleen and stomach qi deficiency and internal accumulation of damp-heat, and giving equal emphasis to eliminating evil and strengthening the body resistance.

[0047] TCM believes that the spleen and stomach is the basis of acquired, the source of qi and blood. The experiment uses the prescription by four gentlemen decoction, composed of six drugs, including ginseng, poria, licorice, dried tangerine or orange peel, patchouli and dandelion, all of which are food and medicine similar traditional Chinese medicine. Ginseng tonifies the qi of five zang organs, poria invigorates the spleen and removes dampness, licorice supplements the middle jiao, removes toxicity and promotes qi, dried tangerine or orange peel dries dampness and promotes qi, patchouli resolves dampness and harmonizes the stomach, and dandelion clears heat and removes toxicity. The whole prescription can invigorate the spleen and stomach, resolve dampness and remove toxicity. TCM treatment of H. pylori infection is flexible and diverse, not limited to the idea of "seeing bacteria and inhibiting bacteria". Food and medicine similar traditional Chinese medicine is a kind of medicine approved by the state, which is both medicine and food. It has the characteristics of high safety, small side effects, no drug resistance, long-term use and easy acceptance by patients. This exactly solves the current problems in the clinical treatment of H. pylori related gastritis. Food and medicine similar traditional Chinese medicine can target the symptoms of H. pylori related gastritis, such as dysfunction of the spleen and stomach, heat and toxic evil, dampness and qi stagnation, and give equal emphasis to eliminating evil and strengthening the body resistance, inhibit the level of inflammatory factors, block the disease and improve the eradication rate of H. pylori.

[0048] Qingyouhugan paste can improve the pathological damage of gastric mucosa in mice with H. pylori related gastritis. Qingyouhugan paste can significantly inhibit the expression of Cag A, a virulence factor of H. pylori in the stomach tissue of mice with H. pylori related gastritis. Qingyouhugan paste can significantly improve the oxidative stress and inflammatory response in mice with H. pylori related gastritis. The concept of TCM prevention and health care has been deeply rooted in the hearts of the people and has become a hot spot of public concern. Food and medicine have the characteristics of high safety, few side effects, suitable for prevention, long-term use and other characteristics, and have obvious advantages in the intervention of aging related diseases. Food and medicine is a unique healthy resource, and the development of food and medicine industry will promote the further development of China's health industry.

[0049] Compared with the prior art, the application has the following beneficial effects:

[0050] The Qingyouhugan paste of the application follows the theory of Chinese dietetics, and has the effects of tonifying qi and spleen and clearing and removing damp heat, and is suitable for the adjuvant treatment of H. pylori related gastritis. The selected Chinese medicines are all from the list of medicinal and edible materials, and are safe and non-toxic. The honey is used to improve the taste of the Qingyouhugan paste, improve the acceptance of the product by the public, and has certain effects and color, aroma, taste and shape as food, and better taste. The medicinal material extraction process is scientific and stable, the production process is simple, and the product has wide market prospects. BRIEF DESCRIPTION OF DRAWINGS

[0051] Figure 1 is a photo of culturing and identifying H. pylori.

[0052] Figure 2 is a photo of HE staining of gastric mucosa of H. pylori.

[0053] Figure 3 is a photo of HE staining of gastric mucosa of mice in different experimental groups in Example 5.

[0054] Among them, NC: control group; MC: model group; LD: low-dose Qingyouhugan paste group; MD: medium-dose Qingyouhugan paste group; HD: high-dose Qingyouhugan paste group; PC: positive drug group.

[0055] Figure 4 is a total ion chromatogram of a blank control solution (negative ion).

[0056] Figure 5 is a total ion chromatogram of a test solution of Qingyouhugan paste extract (negative ion).

[0057] Figure 6 is a total ion chromatogram of a blank control solution (positive ion).

[0058] Figure 7 is a total ion chromatogram of a test solution of Qingyouhugan paste extract (positive ion). DETAILED DESCRIPTION

[0059] The application will be described in detail below in combination with examples. It should be noted that the examples in the application and the features in the examples can be combined with each other without conflict.

[0060] Example 1

[0061] The example provides a Qingyouhugan paste, which mainly comprises:

[0062] Medicinal and edible Chinese medicine ingredients: 6-12 parts of ginseng, 12-24 parts of poria, 9-18 parts of raw licorice, 9-18 parts of patchouli, 9-18 parts of dried tangerine or orange peel, and 15-30 parts of dandelion.

[0063] Food materials: 60-120 parts of honey.

[0064] The preparation process of the Qingyou Hugan Paste is as follows:

[0065] S1, soak the medicinal materials ginseng, poria, raw licorice, patchouli, dried tangerine or orange peel, and dandelion, and measure the water absorption rate.

[0066] S2, use orthogonal test method to investigate the three factors (cooking times, cooking time, and water addition multiple) that affect the extraction effect of the Qingyou Hugan Paste, and optimize the extraction process by taking the dry paste yield as the investigation index.

[0067] S3, mix the first, second, and third extraction liquids, and concentrate under reduced pressure to a clear paste with a relative density of 1.2 (20℃);

[0068] S4, heat the honey in a pot until the temperature reaches 116-118℃, and a large amount of light yellow bubbles appear, and the relative density is about 1.37. Cool and reserve for use.

[0069] S5, add an equal amount of refined honey, continue to concentrate, and stop heating when the "stringing" state appears. The paste collection is completed.

[0070] S6, place the paste obtained in S5 in a high-temperature sterilized ceramic bowl, cool the paste for 12-16 hours, pour it into a high-temperature sterilized glass bottle, and store it in a 4℃ refrigerator.

[0071] Example 2

[0072] This example provides a Qingyou Hugan Paste, which mainly includes:

[0073] Medicinal and edible Chinese medicine ingredients: 6 parts of ginseng, 12 parts of poria, 9 parts of raw licorice, 9 parts of patchouli, 9 parts of dried tangerine or orange peel, and 15 parts of dandelion; food materials: 60 parts of honey.

[0074] The preparation process of the Qingyou Hugan Paste is the same as that of Example 1.

[0075] Example 3

[0076] The embodiment provides a Qingyou Hugan ointment, which mainly comprises

[0077] Medicinal and edible Chinese medicinal ingredient: 12 parts of ginseng, 24 parts of poria, 18 parts of raw licorice, 18 parts of patchouli, 18 parts of dried tangerine or orange peel, and 30 parts of dandelion.

[0078] Food material: 120 parts of honey;

[0079] The preparation process of the Qingyou Hugan ointment is the same as that in Embodiment 1.

[0080] Embodiment 4: Investigation of water extraction process parameters in the preparation process of the Qingyou Hugan ointment of the application

[0081] 1Instrument

[0082] Table 1 Instrument equipment

[0083]

[0084] 2Medicinal materials and reagents

[0085] Ginseng, raw licorice, poria, dried tangerine or orange peel, dandelion and patchouli are purchased from Changsha Haosheng Chinese Herbal Medicine Slice Co., Ltd. (batch numbers are 230901, 231101, 230901, 231101, 231101, 20231101 respectively), and are identified to meet the relevant provisions in the first part of the 2020 edition of Chinese Pharmacopoeia. Octadecylsilane bonded silica gel (Ultimate XB-C18 type chromatographic column, 5 μm, 4.6*250 mm) is purchased from Shanghai Moonstar Technology Co., Ltd. Methanol (chromatographic pure) and acetonitrile (chromatographic pure) are purchased from Thermo Fisher Scientific Company. Methanol (mass spectrometric pure) and acetonitrile (mass spectrometric pure) are purchased from OCEANPAK Company.

[0086] 3Soaking process research

[0087] Ginseng, poria, raw licorice, patchouli, dried tangerine or orange peel and dandelion are weighed according to the prescription amount, 5 parts of medicinal materials and slices are added with 8 times the amount of water, and then soaked at room temperature for 30 min, 60 min, 120 min, 240 min, 480 min and 720 min respectively. The volume of filtered water is measured, and the water absorption rate is calculated. The water absorption rate of medicinal materials = (weight of medicinal materials after water absorption - weight of medicinal materials before water absorption) / weight of medicinal materials before water absorption * 100 / %.

[0088] 4Optimization of extraction process

[0089] The orthogonal test method L9(3)4 was used to investigate the three factors (cooking times, cooking time, and water multiple) affecting the extraction effect of Qingyou Hugan Paste, and the extraction process was optimized with the dry paste yield as the investigation index. According to the prescription amount, 9 parts of traditional Chinese medicine decoction pieces were soaked for 2 hours. According to the test design scheme, decoction was carried out, and the decoction liquid was filtered and concentrated to 60 mL. 60 mL of the extraction liquid in the 250 mL volumetric flask was precisely taken, placed in an evaporating dish dried to a constant weight, and volatilized in a water bath at 105℃ for 3 h. It was moved to a desiccator and cooled for 30 min, and the weight was precisely determined. The dry paste yield was calculated. Dry paste yield % = total weight of solid (g) / total weight of medicinal materials (g) x 100%.

[0090] Table 2 orthogonal design table of extraction process

[0091]

[0092] 5 Ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF / MS) analysis of components in Qingyou Hugan Paste stock solution

[0093] 1) The blank control solution was obtained by centrifuging 12000 r / min for 10 min. The test solution of Qingyou Hugan Paste was prepared by reducing pressure concentration to 1 g / mL, filtering and centrifuging, and finally freeze-drying to dry extract powder. 0.1 g of dry extract powder was precisely weighed and placed in a 10 mL centrifuge tube, and methanol was added to 10 mL. Filter, centrifuge 12000 r / min for 10 min, and take the supernatant to a sample vial.

[0094] 2) Chromatographic conditions: octadecylsilane bonded silica gel as filler, methanol as mobile phase A, 0.1% formic acid aqueous solution as mobile phase B. Column temperature 35℃, flow rate 0.3 mL / min. Gradient elution, gradient elution procedure 0-10 min, 10% A, 10-45 min, 10%→90% A, 45-49 min, 90% A. 49-50 min, 90→10% A, 50-60 min, 10% A.

[0095] 3) Detection by quadrupole time-of-flight mass spectrometer, electrospray ion source (ESI), multiple ion scan of each compound IDA mode parent ion, ion source temperature 500℃. The positive ion mode conditions are first cone hole voltage (80) ± 0V, first collision voltage (10) ± 0V, second cone hole voltage (80) ± 0V, second collision voltage (35) ± (15) V, and the negative ion mode is first cone hole voltage (-80) ± 0V, first collision voltage (-10) ± 0V, second cone hole voltage (-80) ± 0V, second collision voltage (-35) ± (-15) V. The scan mass-to-charge ratio range is 100-1000 amu, the scan gap is 0.661 sec, and the collection time is 0 min and 60 min, respectively. Precise suction of blank control solution 5 μL, supply of test solution 10 μL, injection into liquid chromatograph-mass spectrometer for analysis for 5-45 minutes, and then obtained.

[0096] 6 Paste collection

[0097] Honey has the effects of seasoning and tonifying. Refined honey can remove impurities, kill microorganisms, reduce the water content in the paste, destroy enzymes, increase adhesion, and prolong the shelf life of the paste. The amount of honey used in the paste is generally high, and the ratio of the amount of clear paste to refined honey used during the paste collection process is generally 1:1-1:3. The method of refining honey is to place honey in a pot and heat it to a temperature of 116-118℃, with a large amount of light yellow bubbles appearing and the relative density being about 1.37. The traditional paste concentrate collection is determined by practical experience, such as using the "flag hanging" in summer and "silk hanging" in winter. The extraction solution of the optimized Qingyouhugan Paste is concentrated under reduced pressure to a clear paste with a relative density of 1.2 (20℃), and an equal amount of refined honey is added. Continue to concentrate until the "silk drawing" state appears, stop heating, and the paste collection is completed. Then, the paste is placed in a high-temperature sterilized ceramic bowl, cooled for 12-16 hours, and then poured into a high-temperature sterilized glass bottle and sealed for storage in a 4℃ refrigerator.

[0098] 7 Preliminary quality control study

[0099] 7 Results

[0100] 7.1 Soaking process

[0101] Experiments have proved that the water absorption rate of medicinal materials reaches 103% after soaking for 120 minutes, indicating that the water absorption of medicinal materials has basically reached a saturated state. Therefore, the soaking time of Qingyouhugan Paste is determined to be 120 minutes. The results are shown in Table 3.

[0102] Table 3 Water absorption rate and soaking time

[0103]

[0104] 7.2 Water extraction process optimization results

[0105] Table 4 orthogonal experiment analysis of water extraction process of Qingyouhugan paste

[0106]

[0107]

[0108] Table 5 analysis of variance table

[0109]

[0110] 6.2 verification test

[0111] The results are shown in Tables (4, 5), and the range analysis shows that the factors affecting the extraction effect in order are decoction time > decoction times > liquid ratio. The variance analysis indicates that the effects of A, B, and C on the dry paste yield do not reach a significant level (P>0.05). The decoction time K2 is greater than K1 and K3, so the optimal decoction time is 1 hour; the decoction times K3 is greater than K2 and K1, so the optimal decoction times is 3 times. The water ratio K1 is greater than K2 and K3, and the variance analysis has no statistical difference, so considering the cost saving and dry paste yield, it is appropriate to add 10 times water. Based on the above results, taking the dry paste yield as the evaluation index, the optimal water extraction process of the paste is determined as A2B2C3, i.e. adding 10 times water, extracting for 1 hour each time, and decocting for 3 times.

[0112] 6.3 verification of the optimal extraction process

[0113] According to the optimal process, accurately weigh 6 parts of ginseng, 12 parts of poria, 9 parts of raw licorice, 9 parts of patchouli, 9 parts of dried tangerine or orange peel, and 15 parts of dandelion, soak for 2 hours, add 10 times the amount of water, extract for 1 hour, and decoct for 3 times. Calculate the dry paste yield of 5 parallel samples according to the optimized process. The RSD value of the dry paste yield of the 5 samples extracted according to the optimized process is 1.53%, indicating that the optimized process is stable.

[0114] Table 6 verification results of water extraction process

[0115]

[0116] 6.4 UPLC-Q-TOF / MS analysis of Qingyouhugan paste stock solution composition results

[0117] The total ion chromatograms of the blank control solution and the Qingyouhugan paste extract solution are shown in Figure 4 Under the proposed analysis conditions, 128 components were identified in the Qingyouhugan paste stock solution, including natural product components such as polysaccharides, organic acids, alkaloids, flavonoids, and saponins. The top 20 components in terms of peak area are shown in Table 7.

[0118] Table 7 top 20 components in terms of peak area in the Qingyouhugan paste stock solution

[0119]

[0120] Example 5: Animal experiment of Qingyouhugan Paste of the application

[0121] 1 Experimental instruments and materials

[0122] 1.1 Animals and feed

[0123] 1 Experimental materials

[0124] 1.1 Experimental strains

[0125] Helicobacter pylori standard strain (ATCC43504, North China Biology Technology Co., Ltd., China). The standard strain is also known as NCTC 11637, derived from human antrum, containing Cag A and Vac A.

[0126] 1.2 Experimental animals

[0127] 60 SPF C57 / BL6 mice, half male and half female, 6-7 weeks old, body weight 18-22 g, purchased from Hunan Slike Jingda Experimental Animal Co., Ltd., animal qualification certificate number: 430727231102253251, license number: SCXK(Xiang)2019-0004. The animals were adaptively fed for 7 days, and the environment was kept at room temperature 25℃, humidity 50%~70%. This experiment was approved by the Ethics Committee of Hunan Anshengmei Pharmaceutical Research Institute, and the ethics number was IACUC-KY2023014.

[0128] 1.3 Experimental drugs

[0129] Qingyouhugan Paste obtained in Experiment 2.4.1 was stored in a 4℃ refrigerator. Omeprazole enteric-coated capsules (Changzhou Four Medicine Pharmaceutical Co., Ltd., batch number H10950086); Amoxicillin capsules (ZhuHaiLianBiao Pharmaceutical Co., Ltd., batch number H20003263); Clarithromycin dispersible tablets (Yangtze River Pharmaceutical Group Co., Ltd., batch number H9990376).

[0130] 1.4 Main experimental reagents and consumables

[0131] Table 8 Main experimental reagents

[0132]

[0133]

[0134] 1.3 Main test instruments

[0135] Table 9 Main experimental reagents

[0136]

[0137] 2 Indicators and selection basis

[0138] 2.1 Bacterial culture

[0139] Weigh 3.9 g of Columbia blood agar into a conical flask with 100 mL of distilled water, autoclave at 121 °C for 30 minutes, cool to 40-50 °C, add 5% sterile defibrinated sheep blood, mix well and immediately prepare the blood agar plate. Take out the stored H. pylori strain, dissolve with appropriate amount of sterile PBS (containing 15% glycerol), and all operations related to H. pylori are performed in a biosafety level II laboratory. Use a sterile inoculation loop to inoculate the bacterial solution onto the base culture plate, evenly spread, then invert the blood plate in a three-gas (85% N2, 5% O2, 10% CO2) incubator for culture, and observe the growth of H. pylori after 3-5 days.

[0140] 2.2 Bacterial identification

[0141] 2.2.1 Urease detection

[0142] Use a sterile cotton swab to pick up the colonies on the blood agar plate and add them to the substrate solution for urease detection, seal with a parafilm, and place in a 37 °C environment for 3-5 min. Observe the color change of the reagent, if it changes to pink to rose red, it is positive.

[0143] 2.2.2 Gram staining

[0144] 1) Preparation and fixation: Drop a small amount of PBS buffer in the center of the slide, use a sterile cotton swab to pick up the colonies and repeatedly spread them. Use the flame of an alcohol lamp to fix the bacteria.

[0145] 2) Follow the steps in the instructions to perform the staining.

[0146] 3) Microscopic observation: Air dry the slide and observe under a 1000x optical microscope. The bacteria are stained blue-violet for Gram-positive bacteria and red for Gram-negative bacteria.

[0147] 2.3 Model preparation

[0148] 60 mice were evenly divided into groups according to body weight, with 10 in the control group (NC) and 50 in the model group, half male and half female. After 1 week of adaptive feeding, modeling began. According to the literature, the H. pylori was administered by gavage, and the mice were fasted for 12 h and deprived of water for 4 h before gavage. The model group was first given 0.2 mL of 5% NaHCO3 solution intragastrically to neutralize stomach acid, and 1 h later, 0.2 mL of H. pylori solution (concentration 1 x 10 9CFU / mL)0.3mL, the control group mice were given an equal volume of normal saline gavage, gavage after continuing to fast, water 4h, and then given normal diet. Gavage every other day, a total of 5 times, the last gavage after the fixed value 2 weeks.

[0149] 2.4 Model evaluation

[0150] After the fixed value 2 weeks, the model group randomly selected 10 mice, do Giemsa staining and urease experiment, to test whether the modeling is successful.

[0151] 2.4.1 Urease detection

[0152] A small piece of gastric mucosa near the pylorus of the stomach was cut and placed in a test tube containing urease reagent, sealed with a sealing film, and incubated at 37℃ for 3-5 min. The color change of the microwell strip was observed, and if it changed to light red to rose color, it was positive.

[0153] 2.4.2 Giemsa staining

[0154] After dehydration and embedding of the gastric tissue, the wax block was cut into 4μm sections and placed in hot water to flatten the sections. Then they were placed in an incubator for 1h, then in xylene, and then in 95%, 85%, and 70% ethanol for about 10min each time until the paraffin was completely dissolved. According to the steps of the modified Giemsa staining kit, the sections were stained, air-dried, and analyzed under a light microscope.

[0155] 2.5 Grouping, administration

[0156] After the modeling was successful, the model mice were divided into control group (normal control group, NC), model group (model control group, MC), low dose group of Qingyou Hugan paste (low dose group, LD), middle dose group of Qingyou Hugan paste (middle dose group, MD), high dose group of Qingyou Hugan paste (high dose group, HD), and positive drug group (positive control group, PC). According to the mouse and human equivalent dose conversion formula in Pharmacological Experimental Methods, the gavage amount (g / kg·d) = 9.1 × total amount of human drug (g) / 70. The equivalent dose of Qingyou Hugan paste and positive drug group was finally calculated. Qingyou Hugan paste was given at low, middle, and high doses of 3.9mg / kg, 7.8mg / kg, and 15.6mg / kg, respectively, which was equivalent to 0.5, 1, and 2 times the clinical dose, twice a day, for 14 consecutive days. The positive drug group was given omeprazole (2mg / kg) + amoxicillin (100mg / kg) + clarithromycin (50mg / kg) gavage, twice a day, for 14 consecutive days.

[0157] 2.6 Sample collection

[0158] After the end of administration, the mice in each group were fasted for 12 h without water. The surgical instruments and dressings were sterilized by high-pressure steam sterilization before the operation, and strict aseptic operation was strictly followed. The eyeballs of the mice in each group were removed to collect blood, and the blood was collected in an EP tube and left at room temperature for 1 h, then centrifuged at 3000 r / min for 10 min to separate the serum, which was stored in a -80℃ refrigerator for testing. After the blood sample was collected, the mice were treated by cervical dislocation, and the stomach tissue was collected, fixed in 4% paraformaldehyde solution or stored in a -80℃ refrigerator for standby.

[0159] 2.7 Index detection

[0160] 2.7.1 Observation of the physical signs, symptoms and body weight of mice in each group

[0161] Observe whether the mice in each group have death, body length, hair, diet, spirit, and activity, and record the body weight changes every week.

[0162] 2.7.2 Observation of the pathological structure of the gastric mucosa of mice in each group

[0163] 1) Dehydration and embedding: after being fixed in 4% paraformaldehyde for 24 h, the tissue was taken out, dehydrated with alcohol (from low concentration to high concentration), and then placed in xylene. The treated stomach tissue was placed in melted paraffin, and the embedded wax block was quickly placed in a refrigerator for cooling.

[0164] 2) Sectioning and patching: the wax block was cut into 4 μm sections, placed in hot water to flatten, transferred to a glass slide, dried in a constant temperature oven, and sealed and stored in a 4℃ refrigerator.

[0165] 3) De-waxing and hydration: the section was placed in a constant temperature oven for 1 h, then placed in xylene, and the xylene was replaced and continued to soak; sequentially placed in 95%, 85%, and 70% ethanol for about 10 min each time, until the paraffin was completely dissolved.

[0166] 4) HE staining: the hydrated section was sequentially washed with distilled water for 3 times; transferred to hematoxylin for 3 min and tap water for 5 min. Transferred to differentiation solution, the differentiation time was 3 sec, the tissue section was faded to light blue red, then washed with water for 15 min, and transferred to eosin staining solution for 2 min; the dehydrated sample section was soaked in xylene for 2 times, 5 min each time; and sealed with neutral balsam.

[0167] 2.7.3 Detection of Cag A mRNA expression in the stomach tissue of mice in each group by RT-PCR

[0168] 1) Take 50 mg of each group of mouse stomach tissue from the ultra-low temperature refrigerator for frozen preservation and place it in a 1.5 mL sterile enzyme-free centrifuge tube. Extract total RNA according to the instructions.

[0169] 2) Use a microspectrophotometer to measure A260, A280 and A260 / A280, and calculate the purity and concentration of RNA.

[0170] 3) Prepare the reverse transcription reaction system according to the reverse transcription kit, mix and centrifuge, then incubate at 50°C for 15 min, 85°C for 5 sec, and terminate the reaction. Store the extracted cDNA in a -80°C refrigerator or immediately perform RT-PCR detection.

[0171] 4) Configure the reaction system according to the SYBR Green qPCR kit. Complete amplification on a fluorescence quantitative PCR instrument, and the reaction conditions are as follows: ① pre-denaturation: 95°C, 5 min, cycle 1; ② denaturation: 95°C, 10 sec, cycle 40; ③ annealing: 58°C, 20 sec, cycle 40; ④ extension: 72°C, 20 sec, cycle 40.

[0172] 5) The final data is analyzed by 2-ΔΔCt, and the primers are designed by Hunan Aikuerui Biological Engineering Co., Ltd. The sequences are shown in the following table.

[0173] Table 10 Primer Sequence Table

[0174]

[0175] 2.7.4 ELISA method for detecting IL-8, IL-10, TNF-α expression in serum of each group of mice

[0176] The content is detected according to the method in the ELISA kit instructions. The specific steps are as follows: set standard wells, sample wells and blank control wells. Add serum according to the kit instructions, add horseradish peroxidase HRP labeled antibody to the standard wells and sample wells, and only add PBS buffer to the blank wells. After covering and sealing, incubate in a 37°C incubator for 60 min. Discard the reaction solution, wash the plate 5 times (1 min each time). Add substrate A and B to each well in turn, incubate in a water bath for 15 min, then add stop solution. Within 30 minutes, use a microplate reader to measure the absorbance of each well (wavelength 450 nm), and calculate the sample concentration.

[0177] 2.7.4 ELISA method for detecting IL-18, IL-1β expression in stomach tissue of each group of mice

[0178] The content was detected according to the method in the ELISA kit instruction. The specific steps are as follows: set standard hole, sample hole and blank control hole. Add gastric tissue according to the kit instruction, add horseradish peroxidase HRP labeled antibody in standard hole and sample hole, and only add PBS buffer in blank hole. After covering film and sealing, incubate in 37℃ incubator for 60min. Discard the reaction solution, wash the plate 5 times (1min / time). Add substrate A and B to each hole in turn, incubate in water bath for 15min, and then add stop solution. Within 30min, use enzyme label instrument to detect the absorbance of each microwell (wavelength 450nm), and calculate the sample concentration.

[0179] 2.7.5 SOD and MDA detection of gastric tissue of mice in each group

[0180] 1) Protein extraction: take 20mg of mouse gastric mucosa, wash with ice-cold PBS for 2-3 times, and move to EP tube. Add 200μl of pre-cooled lysis buffer (add twice), mechanically cut the tissue on ice, and oscillate for 10min intermittently, then homogenize on ice until complete lysis. Centrifuge at 4℃, 12000r / min for 10min, and collect the supernatant as total protein sample.

[0181] 2) BCA protein concentration determination: prepare BCA working solution according to the proportion of A and B liquid 50:1, and mix well. Dilute 5mg / mL BSA standard solution with PBS to 1.0mg / mL, and then dilute to obtain 0, 62.5, 125, 250, 500 and 1000μg / mL protein standards, and take 20μL of each into the microwell plate. Dilute the sample at a ratio of 1:20 (1μL sample+19μL PBS), and take 20μL of each into the 96-well plate, and add 200μl of BCA working solution to each well. After 37℃ dark reaction for 30min, measure the absorbance at 562nm with enzyme label instrument, and calculate the protein concentration through the standard curve.

[0182] 3) WST-1 method for detecting gastric mucosa SOD. The specific steps are as follows:

[0183] ① Homogenate preparation: weigh 100mg of mouse gastric tissue, add 9 times the volume of normal saline, cut the tissue, and prepare homogenate in ice water bath, 2500-4000r / min, frozen centrifugation for 10min, and take the supernatant as 10% homogenate supernatant.

[0184] ② Prepare each reagent according to the kit requirements, and the operation is as follows:

[0185] Table 11 SOD detection working solution configuration parameters

[0186]

[0187] ③ Mix well, incubate at 37℃ for 20min, and measure the OD value at 450nm with enzyme label instrument.

[0188] SOD inhibition rate (%) =

(control OD value - control blank OD value) - (determination OD value - determination blank OD value)

[0189] SOD activity (U / mg) = SOD inhibition rate / 50%*reaction system dilution ratio (0.24 mL) / (0.02 mL) / protein concentration of sample to be tested (mg / mL)*dilution ratio before sample test

[0190] 4) TBA method for detecting MDA level in gastric mucosa tissue

[0191] Table 12 MDA detection working solution configuration

[0192]

[0193] According to the instruction process, the OD value at 532 nm of the microplate reader. MDA content (nmol / mg prot) = (determination OD value - control OD value) / (standard OD value - blank OD value) x standard concentration (10 nmol / mL) / sample protein concentration.

[0194] 2.8 Statistical analysis

[0195] SPSS 27.0 statistical software was used for processing and analysis. If the measurement data conforms to the normal distribution, it is represented by mean ± standard deviation, and the comparison between multiple groups uses one-way ANOVA, and the LSD test is used for variance homogeneity post-test, and the Tamhane's T2 test is used for variance inhomogeneity. If it does not conform to the normal distribution, it is statistically analyzed by Kruskal-Wallis H rank sum test. The test level is taken as α = 0.05, and P < 0.05 is statistically different.

[0196] 3 Experimental results

[0197] 3.1 Helicobacter pylori identification results

[0198] Helicobacter pylori was cultured on solid medium for 3-4 days, showing transparent colonies of needle size (A). Urease can quickly decompose urea to produce NH3 and CO2, and the rapid urease test shows a light red to rose red change, which is positive (B); the grown Helicobacter pylori colonies were picked and stained by Gram staining, and identified as Gram-negative bacteria (C) Figure 1 Figure 1 Figure 1

[0199] 3.2 Model evaluation results

[0200] As Figure 2 ​​​The urease staining showed pink to rose red color change, indicating positive (A), and the model group showed local ulceration in the gastric mucosa layer, unclear structure of the mucosa layer, and a large number of Helicobacter pylori in the ulcer focus, showing blue color and short rod shape (B).

[0201] 3.3 Influence of Qingyouhugan Paste on the signs, symptoms and body weight of mice in each group

[0202] Through experimental observation, the control group mice were active, had bright eyes, and had good appetite. The hair was shiny. On the third day of modeling, the mice began to have decreased appetite and reduced activity. During the continued modeling period, the hair gradually lost its luster, the animals were tired and liked to huddle, the activity was reduced, the food intake was poor, and the body weight increased slowly. After 2 weeks of drug intervention, some animals in the low-dose group of Qingyouhugan Paste still had tired expression and liked to huddle, the animals in the medium and high-dose groups had shiny hair, increased activity, and increased food intake. The animals in the positive drug group had increased activity and increased food intake.

[0203] The body weight of mice in each group was observed and recorded. After 2 weeks of drug intervention, the results showed that compared with the control group, the body weight of mice in the model group was significantly decreased (P<0.05). After intervention with Qingyouhugan Paste and positive drugs, the body weight of mice in each group showed a gradually increasing trend. Compared with the model group, the differences in the medium and high-dose groups and the positive drug group were statistically significant (P<0.05). Compared with the high-dose group, the body weight of mice in the low-dose group was different (P<0.05), and there was no significant difference in the body weight of mice in the medium-dose group (P>0.05). Compared with the positive drug group, the body weight of mice in the low-dose group was different (P<0.05), and there was no significant change in the body weight of mice in the medium and high-dose groups (P>0.05).

[0204] Table 13 Body weight of mice in each group after 14 days of drug administration

[0205]

[0206] Note: The data is represented by mean ± standard deviation (n=6). NC: control group; MC: model group; LD: low-dose Qingyouhugan Paste group; MD: medium-dose Qingyouhugan Paste group; HD: high-dose Qingyouhugan Paste group; PC: positive drug group. # P<0.05, compared with NC; * P<0.05, compared with MC; △ P<0.05, compared with HD; ○ P<0.05, compared with PC.

[0207] 3.4 Influence of Qingyouhugan Paste on Cag A mRNA in the gastric tissue of mice in each group

[0208] Table 14 Effect of Qingyouhugan Paste on Cag A mRNA of each group of mice

[0209]

[0210] Note: Data is expressed as mean ± standard deviation, (n = 3). NC: control group; MC: model group; LD: Qingyouhugan Paste low-dose group; MD: Qingyouhugan Paste medium-dose group; HD: Qingyouhugan Paste high-dose group; PC: positive drug group. # P<0.05, compared with NC; * P<0.05, compared with MC; △ P<0.05, compared with HD; ○ P<0.05, compared with PC.

[0211] The experimental results suggest that compared with the control group, the expression level of Cag A mRNA in the stomach tissue of the model group of mice increased (P<0.05); compared with the model group, the expression level of Cag A mRNA in the stomach tissue of the medium and high dose groups of Qingyouhugan Paste and the positive drug group decreased (P<0.05). Compared with the high dose group, the expression level of Cag A mRNA in the stomach tissue of the medium and low dose groups was different (P<0.05). Compared with the positive drug group, the expression level of Cag A mRNA in the stomach tissue of the high dose group of Qingyouhugan Paste decreased, and the difference was statistically significant (P<0.05).

[0212] 3.5 Effect of Qingyouhugan Paste on the pathological structure of the gastric mucosa of each group of mice

[0213] HE Figure 3 staining showed that in the control group of mice, the gastric mucosa epithelium was complete, the columnar epithelial cells were arranged in order, the gastric glands in the lamina propria were arranged in order and closely, the chief cells and parietal cells had no obvious abnormalities, the connective tissue in the submucosa was relatively fine and dense, there was no obvious edema, the muscular layer had no obvious abnormalities, there was no obvious ulcer, and there was no obvious inflammatory cell infiltration. In the model group of mice, superficial ulcers were visible in the gastric tissue, the mucosa layer was thin, the mucosa epithelial cells were necrotic and exfoliated, accompanied by neutrophil infiltration, and occasionally basophilic granular substances were seen, suspected to be bacteria, and scattered inflammatory cell infiltration was visible in the local submucosa. In the Qingyouhugan Paste dose groups, no obvious ulcers were visible, the gastric mucosa epithelium was complete, the columnar epithelial cells were arranged in order, the gastric glands in the lamina propria were arranged in order and closely, and scattered inflammatory cell infiltration was visible in the local submucosa. In the positive drug group, the gastric mucosa epithelium was complete, the columnar epithelial cells were arranged in order, the gastric glands in the lamina propria were arranged in order and closely, the chief cells and parietal cells had no obvious abnormalities, a small amount of inflammatory cell infiltration was visible in the local submucosa, and the muscular layer had no obvious abnormalities.

[0214] 3.6 Effect of Qingyouhugan Paste on IL-8, IL-10, and TNF-α in the serum of each group of mice

[0215] Table 15 Effect of Qingyouhugan Paste on IL-8, IL-10, TNF-α in serum of each group of mice

[0216]

[0217]

[0218] Note: Data are expressed as mean ± SD, (n = 3). NC: control group; MC: model group; LD: Qingyouhugan Paste low-dose group; MD: Qingyouhugan Paste middle-dose group; HD: Qingyouhugan Paste high-dose group; PC: positive drug group. # P < 0.05, compared with NC; * P < 0.05, compared with MC; △ P < 0.05, compared with HD; ○ P < 0.05, compared with PC.

[0219] The experimental results (Table 15) suggest that compared with the control group, the IL-8 and TNF-α in serum of the model group of mice increased (P < 0.05); compared with the model group, the IL-8 level in serum of the Qingyouhugan Paste middle-dose group and high-dose group and the positive drug group decreased, with statistical significance (P < 0.05), and the TNF-α in the Qingyouhugan Paste groups and the positive drug group decreased to different degrees (P < 0.05). Compared with the high-dose group, the IL-8 and TNF-α in serum of the low-dose group of mice had differences (P < 0.05), and the middle-dose group had no significant difference (P > 0.05). Compared with the positive drug group, the TNF-α in serum of the low-dose group of mice had differences (P < 0.05), and the middle-dose and high-dose groups had no significant changes (P > 0.05). Compared with the positive drug group, the IL-8 level in serum of the Qingyouhugan Paste groups had no significant changes (P > 0.05). Compared with the control group, the IL-10 in serum of the model group of mice decreased significantly (P < 0.05); compared with the model group, the IL-10 in serum of the Qingyouhugan Paste groups and the positive drug group increased to different degrees (P < 0.05). Compared with the high-dose group, the IL-10 content in serum of the middle-dose group had no significant changes (P > 0.05), and the low-dose group had differences (P < 0.05). Compared with the positive drug, the IL-10 in serum of the low-dose group had differences (P < 0.05). The experimental results show that Figure 3IL-18, IL-1β in serum of mice in model group were significantly higher than those in control group (P<0.05); IL-18, IL-1β in serum of mice in each dose group of Qingyouhugan Paste and positive drug group were decreased to different degrees compared with model group (P<0.05). Compared with high dose group, IL-1β in serum of mice in middle dose group was decreased (P<0.05), and there was no significant change in low dose group (P>0.05). Compared with high dose group, IL-18 in serum of mice in low dose group was different (P<0.05), and there was no significant change in middle dose group (P>0.05). Compared with positive drug group, IL-18, IL-1β in serum of mice in middle dose group were different (P<0.05), and there was no significant change in low and high dose groups (P>0.05).

[0220] 3.7 Effect of Qingyouhugan Paste on SOD and MDA in gastric tissue of mice in each group

[0221] Table 16 Effect of Qingyouhugan Paste on SOD and MDA in gastric tissue of mice in each group

[0222]

[0223] Compared with control group, SOD in gastric tissue of mice in model group was significantly decreased (P<0.05); compared with model group, SOD in gastric tissue of mice in high dose group of Qingyouhugan Paste and positive drug group was significantly increased (P<0.05). Compared with high dose group of Qingyouhugan Paste, SOD in gastric tissue of mice in middle and low dose groups was different (P>0.05). Compared with positive drug group, SOD in gastric tissue of mice in each dose group of Qingyouhugan Paste was not statistically significant (P>0.05). Compared with control group, MDA in gastric tissue of mice in model group was significantly increased (P<0.05); compared with model group, MDA in gastric tissue of mice in middle and high dose groups of Qingyouhugan Paste and positive drug group was decreased (P<0.05), and there was no difference in MDA in gastric tissue of mice in middle and low dose groups compared with high dose group (P>0.05). Compared with positive drug group, MDA in gastric tissue of mice in low, middle and high dose groups of Qingyouhugan Paste was different (P<0.05). There was no significant change in positive drug group (P>0.05).

[0224] 4 Results

[0225] Through experimental observation, the mice in the control group were active, had bright eyes, and had good appetite. The hair was shiny. On the third day of modeling, the mice began to have decreased appetite and reduced activity. During the continued modeling, the hair gradually lost its luster, the expression was tired, and the animals liked to huddle. The activity was reduced, the food intake was poor, and the body weight increased slowly. After 2 weeks of drug intervention, some animals in the low-dose group of Qingyouhugan Paste still had tired expression and liked to huddle, while the animals in the medium and high-dose groups and the positive drug group had shiny hair, increased activity, and increased food intake. After 2 weeks of drug intervention, compared with the control group, the body weight of the mice in the model group decreased, which may be related to the infection of Helicobacter pylori and the damage to the gastric mucosa. The body weight of the mice in the Qingyouhugan Paste groups increased, indicating that the paste can repair the gastric mucosa and promote the recovery of the mice's signs, symptoms, and body weight.

[0226] The gastric mucus barrier is the first line of defense of the gastric mucosal protective barrier. It and the bicarbonate secreted by the epithelial cells on the surface of the gastric mucosa together form the mucus-bicarbonate barrier, which antagonizes the reverse diffusion of hydrogen ions and the digestion of digestive enzymes, and participates in the defense against damage factors and the repair after damage. The gastric mucosal epithelial cells are in a dynamic process of continuous growth, migration, and shedding, which helps to maintain the integrity of the gastric mucosal epithelium. The experiments showed that compared with the control group, the gastric tissue of the model group mice had superficial ulcers, thin mucosa, necrosis and shedding of mucosal epithelial cells, neutrophil infiltration, and occasional basophilic granular material suspected to be bacteria. Local submucosal layer showed scattered inflammatory cell infiltration. After treatment with Qingyouhugan Paste, no obvious ulcer was observed in each dose group, the gastric mucosal epithelium was complete, the columnar epithelial cells were arranged in order, the gastric glands in the lamina propria were arranged in order and closely, and only scattered inflammatory cell infiltration was observed in the submucosal layer. The pathological damage was recovered, and the results showed that Qingyouhugan Paste could improve the structural damage caused by inflammatory infiltration of the gastric mucosa of HPAG mice to a certain extent from the morphological point of view.

[0227] Helicobacter pylori virulence factors are transported into the host's gastric mucosal cells through a needle-like structure. Activation of NF-κB can lead to the release of pro-inflammatory cytokine interleukin IL-8. IL-8 recruits macrophages, neutrophils and basophils to the site of infection to play an anti-inflammatory role. After invading the host's gastric mucosal cells, H. pylori can activate the recombinant myeloid differentiation factor 88 (MyD88) and tumor necrosis factor (TNF) receptor 1 signaling pathway, inducing macrophages to secrete IL-1, IL-6, IL-12, TNF-α and interferon-γ, and pro-inflammatory cytokines can induce ROS production, further exacerbating H. pylori-induced inflammation and gastric mucosal damage, and excessive ROS production can also lead to mitochondrial dysfunction. Macrophages can phagocytose and destroy pathogens, and as antigen-presenting cells, these cells are usually polarized into classically activated macrophages (M1) and immunoregulatory macrophages (M2). M1 macrophages produce high inducible nitric oxide synthase and pro-inflammatory molecules such as TNF-α, IL-6, IL-12, IL-1β, IL-18, while M2 macrophages produce IL-10, transforming growth factor-β (TGF-β), vascular endothelial growth factor, epidermal growth factor, etc.

[0228] The experiment found that the Cag A mRNA of the model group mice was significantly increased by detecting the Cag A mRNA of the gastric mucosa tissue of HPAG mice. After the intervention of Qingyou Hugan Paste, the content of Cag A mRNA decreased, suggesting that the mechanism of Qingyou Hugan Paste in reducing the gastric mucosal damage of HPAG mice may be related to the inhibition of the expression of H. pylori virulence factors. The ELISA results of this study showed that the pro-inflammatory cytokines TNF-α and IL-8 in the serum of HPAG mice increased to varying degrees, and the immunosuppressive factor IL-10 decreased, while Qingyou Hugan Paste treatment significantly reversed the increase of TNF-α and IL-8, and increased the level of IL-10. Therefore, we speculate that the mechanism of Qingyou Hugan Paste in repairing the gastric mucosal damage of HPAG may rely on the inhibition of the release of pro-inflammatory cytokines, the increase of IL-10 level, and the slowing down of the inflammatory process, thereby playing a protective role.

[0229] Oxidative stress (OS) is closely related to acute gastric mucosal injury. The accumulation of reactive oxygen species (ROS) can have a negative impact on antioxidant enzymes such as superoxide dismutase (SOD) and reduced glutathione (GSH), leading to oxidative stress damage to cells. SOD is a highly active antioxidant enzyme that can reduce neutrophil-mediated inflammatory reactions and inhibit the release of ROS and chemotactic factors from endothelial cells damaged by inflammatory mediators. Malondialdehyde (MDA) is a product of lipid peroxidation, mainly derived from the peroxidation of saturated fatty acids, and is an important biomarker of oxidative stress. After Helicobacter pylori infection of the gastric mucosa, the level of ROS increases, causing oxidative stress, and the increase in the level of ROS can cause gastric mucosal injury, delay the healing of gastric mucosal tissue, and produce precancerous lesions of the gastric mucosa. By detecting the oxidative stress indicators of the gastric mucosa of HPAG mice, it was found that the SOD level of the gastric tissue of the model group mice was significantly lower than that of the control group, and the MDA content was significantly increased. After the intervention of Qingyou Hugan Paste, the SOD level of the gastric tissue of the mice was increased, and the MDA content was decreased, suggesting that the mechanism by which Qingyou Hugan Paste improves the gastric mucosal injury of HPAG mice may be related to the regulation of oxidative stress.

Claims

1. A medicinal and edible composition having the function of treating Helicobacter pylori-related gastritis, characterized in that: By weight, the medicine-food composition includes 6 to 12 parts of ginseng, 12 to 24 parts of Poria cocos, 9 to 18 parts of raw licorice, 9 to 18 parts of patchouli, 9 to 18 parts of tangerine peel, and 15 to 30 parts of dandelion.

2. The medicine-food composition according to claim 1, further comprising 60-120 parts by weight of honey.

3. A method for preparing the medicinal and edible composition for treating Helicobacter pylori-related gastritis as claimed in claim 2, comprising the following steps: S1. Soak ginseng, poria cocos, raw licorice, patchouli, tangerine peel, and dandelion; S2, mixing the ginseng, poria, raw licorice, patchouli, tangerine peel, and dandelion soaked in S1 and extracting with water; S3, mixing the extracts obtained by water extraction in S2, and concentrating to obtain a clear paste; S4, heating the honey, concentrating it, and cooling it to obtain refined honey; S5. Add the refined honey prepared in S4 to the clear paste prepared in S3 and continue to concentrate.

4. The preparation method according to claim 3, characterized in that In step S1, the soaking time is at least 120 minutes.

5. The preparation method according to claim 3, characterized in that In step S2, the decoction material-liquid ratio is 1:(8-15), the decoction time is 1-3 hours, and the decoction is performed 2-4 times.

6. The preparation method according to claim 4, characterized in that In step S3, the relative density of the clear paste is 1.1-1.

3.

7. The preparation method according to claim 2, characterized in that In step S4, the honey is placed in a pot and heated to a temperature of 116° C. to 118° C., a water content of 14% to 16%, and a relative density of 1.2 to 1.

4.

8. Use of the edible and medicinal composition of claims 1 to 2, or the edible and medicinal composition prepared by the preparation method of any one of claims 3 to 7, in preparing an edible and medicinal preparation for treating Helicobacter pylori-associated gastritis.

9. The application according to claim 8, characterized in that: The medicine-food combination is used in preparing a preparation for reversing the decrease of local pro-inflammatory factors IL-18 and / or TNF-α, and / or the increase of IL-10 in enteritis.

10. The use according to claim 8, characterized in that: The medicine-food combination is used in preparing a preparation for reversing the increase of SOD and / or the decrease of MDA, a local oxidative stress factor in enteritis.

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