Traditional Chinese medicine composition for treating chronic atrophic gastritis

Abstract

The composition provided by the invention takes bear gall wood as a monarch drug and takes earthworm, bletilla striata, bighead atractylodes rhizome, Chinese yam and white paeony root as ministerial drugs, the earthworm and bletilla striata jointly play the effects of promoting blood circulation to remove meridian obstruction, dispersing blood stasis, removing stasis, astringing to stop bleeding and repairing injury, and the bighead atractylodes rhizome and Chinese yam are used as ministerial drugs and are used for strengthening spleen and stomach, replenishing qi and blood, providing fundamental power for the whole prescription and supporting healthy qi; liver softening, spasm relieving and pain relieving can be achieved; meanwhile, the traditional Chinese medicine composition has the effects of nourishing blood and astringing yin, can prevent other medicines in the formula from being too warm and dry or consuming yin blood, and forms dynamic and static combination compatibility with the minister medicine earthworm for promoting blood circulation to remove meridian obstruction; hairyvein agrimony, herba violae, vernonia graveolens, cuttlebone, mangnolia officinalis, semen raphani and endothelium corneum gigeriae A large number of pathological experiments prove that the traditional Chinese medicine composition can reduce stomach tissue inflammation, oxidative stress and activate apoptosis signal channels by acting on MAPK, PI3K-Akt and HIF-1 signals, so that the effect of treating chronic atrophic gastritis is achieved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of traditional Chinese medicine, and particularly relates to a traditional Chinese medicine composition for treating chronic atrophic gastritis and a preparation method thereof. BACKGROUND

[0002] Chronic atrophic gastritis (CAG) is a common chronic gastritis clinically, which refers to a chronic gastric disease that the gastric mucosa epithelium suffers from repeated damage to cause the decrease of the intrinsic glands, the thinning of the gastric mucosa, the thickening of the mucosal base, and the intestinal metaplasia and / or pseudo-pyloric metaplasia with or without. The incidence and detection rate thereof increase with age. Helicobacter pylori infection is the most important cause of CAG. The clinical manifestations of CAG are non-specific, and there can be no obvious symptoms. The main symptoms of the patients with symptoms are non-specific dyspeptic symptoms such as upper abdominal discomfort, fullness, pain, and the like, which can be accompanied by loss of appetite, rumbling, belching, acid regurgitation, nausea, bitter mouth, and other digestive tract symptoms. There is no correlation between the severity of the pathology of CAG and the symptoms. The diagnosis of CAG mainly depends on endoscopy and histological examination of the gastric mucosa, and the diagnostic value of the latter is greater. The detection of serum pepsinogen I, II and gastrin-17 is helpful to determine whether there is gastric mucosal atrophy and atrophic site. For the gastric body atrophy, the serum PG I, PG I / II ratio is reduced, and the serum Gastrin-17 level is increased. For the antral atrophy, the serum Gastrin-17 level is reduced, and the serum PG I, PG I / II ratio is normal. For the whole gastric atrophy, both of them are reduced. Usually, PG I level ≤70 g / L and PG I / II ratio ≤3.0 are used as the diagnostic critical value of atrophic gastritis. According to the Correa (Correa) cascade reaction hypothesis, the occurrence of gastric cancer experiences chronic non-atrophic gastritis, atrophic gastritis (atrophy, intestinal metaplasia), dysplasia and cancer.

[0003] At present, the western medicine lacks effective treatment for CAG, and mainly focuses on the helicobacter pylori eradication treatment and endoscopic monitoring management. The eradication of helicobacter pylori cannot significantly improve the antral mucosal atrophy or intestinal metaplasia. The treatment goal is to delay or block the progression of the lesion, reduce the risk of canceration, and improve the clinical symptoms of the patients. There is still lack of exact and effective treatment for reversing the pathological changes of CAG.

[0004] Traditional Chinese medicine has unique advantages in the treatment of digestive system diseases, especially for the pathogenesis of CAG patients. Therefore, through the individualized and dynamic follow-up treatment of traditional Chinese medicine, the level and rate of gastric mucosal repair are improved, and the mucosal pathology of CAG patients is improved. It can undoubtedly provide an effective and characteristic treatment for the population with high helicobacter pylori infection rate and high incidence of gastric cancer in China, supplement the treatment scheme of CAG, and has important significance for the prevention and treatment of gastric cancer.

[0005] CAG in traditional Chinese medicine belongs to "stomach pain", "fullness", "hiccups" and other categories, although the disease is not the same, but the cause of the disease, not only improper diet, irregular work and rest, the body is weak, exogenous six evils, emotional unsmooth and so on. The pathogenesis is mainly in the stomach, and is closely related to the liver and spleen. Because the gallbladder is attached to the liver, and the liver is mainly for the purpose of excretion, so it is also related to the gallbladder; Because CAG has a long course, repeated attacks, and long-term disease, often showing deficiency in the root and excess in the branch, and deficiency and excess mixed syndrome. The main deficiency is spleen and stomach deficiency, and the stomach yin deficiency is mainly; The evil is mainly qi stagnation, blood stasis, dampness, heat and liver depression. In the development process of the disease, spleen and stomach weakness and qi stagnation and blood stasis often cause and effect each other, and appear alternately throughout the whole disease. The liver and stomach qi stagnation syndrome and the spleen and stomach weakness syndrome (spleen and stomach cold syndrome) are the most common, and can also appear together. On the basis of distinguishing single syndrome, the compound syndrome should be distinguished. With the passage of time, the syndrome may change dynamically and need to be carefully identified. It is urgent to develop a general and effective prescription or traditional Chinese medicine preparation for the treatment of patients and market circulation. SUMMARY

[0006] The application provides a traditional Chinese medicine composition for treating chronic atrophic gastritis and a preparation method thereof. The traditional Chinese medicine composition can reduce gastric tissue inflammation, oxidative stress and activate apoptosis signaling pathway by acting on MAPK, PI3K-Akt and HIF-1 signal, so as to achieve the effect of treating chronic atrophic gastritis.

[0007] The application provides a traditional Chinese medicine composition for treating chronic atrophic gastritis, which comprises the following raw materials in parts by weight: 10-20 parts of Aralia chinensis, 10-20 parts of Earthworm, 10-30 parts of Bletilla striata, 10-30 parts of Atractylodes macrocephala, 10-20 parts of Paeonia lactiflora, 10-30 parts of Dioscorea opposita, 10-30 parts of Agrimonia pilosa, 10-20 parts of Viola yedoensis, 5-15 parts of Disporum smilaceifolium, 5-15 parts of Gigeria sinensis, 5-15 parts of Magnolia officinalis, 10-20 parts of Concha arcae, and 10-30 parts of Raphanus sativus.

[0008] Preferably, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 16 parts of Aralia chinensis, 16 parts of Earthworm, 20 parts of Bletilla striata, 20 parts of Atractylodes macrocephala, 15 parts of Paeonia lactiflora, 20 parts of Dioscorea opposita, 20 parts of Agrimonia pilosa, 15 parts of Viola yedoensis, 9 parts of Disporum smilaceifolium, 9 parts of Gigeria sinensis, 9 parts of Magnolia officinalis, 15 parts of Concha arcae, and 20 parts of Raphanus sativus.

[0009] Further, the traditional Chinese medicine composition comprises 20-40 parts or 30 parts of honey.

[0010] Further, the composition is prepared from the active ingredients of the medicinal powder of the raw material or the water or organic solvent extract of the raw material, and the pharmaceutically acceptable adjuvant.

[0011] The application also provides a use of the Chinese medicine composition in the above description in the preparation of a drug for treating chronic atrophic gastritis.

[0012] The bear gall wood in the application is rich in gum. After the bear gall wood is stripped from the tree, a soldering iron or other means is used to seal the broken part of the bark, so as to seal a large amount of gum in the bear gall wood. The present inventors have found through a large number of clinical experiments that the bear gall wood treated in this way is used to prepare the Chinese medicine composition in the application, and unexpected technical effects are achieved.

[0013] Bear gall wood: also known as iron holly, bitter. It belongs to the lung, liver and heart meridians, and has the effects of activating blood, unblocking collaterals, removing blood stasis, clearing heat and resolving toxins, and cooling blood and stopping bleeding.

[0014] Earthworm: cold in nature, salty in taste. It belongs to the liver, spleen and bladder meridians. It has the effects of clearing heat, calming shock, dredging collaterals, relieving asthma, and diuresis. The main components include earthwormin, earthworm toxin, earthworm heat-dissolving substance, various amino acids, xanthine, adenine, guanine, as well as choline, fatty acids, lipids, nucleic acid derivatives, free amino acids, various vitamins, inorganic salts, etc. Modern pharmacological studies have shown that earthworm has the effects of sedation and hypnosis, anti-convulsion, anti-arrhythmia, heat-dissolving, hypothermia, hypotension, influence on blood rheology, anti-platelet aggregation, anti-brain ischemia, anti-coagulation and anti-thrombosis, excitation of gastrointestinal smooth muscle, asthma relief, excitation of uterus, immune regulation, anti-bacterial, anti-tumor, and promotion of wound healing.

[0015] White yam: bitter, sweet and astringent in taste, slightly cold in nature, and belongs to the lung, liver and stomach meridians. It has the effects of astringing and stopping bleeding, and promoting tissue regeneration.

[0016] White atractylodes: bitter and sweet in taste, warm in nature, and belongs to the spleen and stomach meridians. It has the effects of invigorating the spleen and tonifying qi, drying dampness and promoting urination, stopping sweating, and preventing miscarriage.

[0017] White peony root: slightly cold in nature, bitter and sour in taste. It belongs to the liver and spleen meridians. It has the effects of warming yang and expelling dampness, tonifying deficiency, invigorating the spleen and stomach, nourishing blood and regulating menstruation, soothing the liver and relieving pain, and calming the liver yang.

[0018] Chinese yam: sweet in taste, neutral in nature, and belongs to the spleen, lung and kidney meridians. It has the effects of tonifying the spleen and stomach, nourishing the lung, and tonifying the kidney. It is used for treating spleen deficiency, chronic diarrhea, lung deficiency, kidney deficiency, hematochezia, leukorrhagia, and diabetes. Fried Chinese yam is used to tonify the spleen and stomach.

[0019] Herba agrimoniae: bitter and astringent in taste, neutral in nature, and belongs to the heart and liver meridians. It has the effects of astringing and stopping bleeding, stopping diarrhea, and detoxifying. It is used for treating hemoptysis, hematemesis, metrorrhagia, malaria, dysentery, debilitation, sore and toxic, and leukorrhagia.

[0020] Herba violae: bitter and pungent in taste, cold in nature, and belongs to the heart and liver meridians. It has the effects of clearing heat and resolving toxins, and cooling blood and swelling.

[0021] Over-the-mountain dragon: It has a pungent taste and is hot in nature. It enters the heart and kidney meridians and has the effects of dispelling wind and dampness, dispersing blood stasis and reducing swelling.

[0022] Cuttlebone: It is warm in nature and salty and astringent in taste. It enters the spleen and kidney meridians and has unique effects of astringing and stopping bleeding, astringing essence and stopping leukorrhea, neutralizing acid and relieving pain, and astringing dampness and healing sores.

[0023] Chicken gizzard lining: neutral in nature and sweet in taste. It enters the spleen, stomach, small intestine, and bladder meridians, and has the functions of strengthening the stomach and promoting digestion, astringing essence and stopping seminal emission.

[0024] Magnolia officinalis: It tastes bitter and pungent, is warm in nature, and enters the spleen, stomach, lung, and large intestine meridians. It has the effects of drying dampness and resolving phlegm, and relieving fullness by lowering qi.

[0025] Radish seeds: neutral in nature, pungent and sweet in taste. They enter the lung, spleen and stomach meridians, and have the effects of lowering qi and relieving asthma, promoting digestion and resolving phlegm, treating cough with phlegm and wheezing, food stagnation and qi stagnation, chest tightness and abdominal distension, and tenesmus after diarrhea.

[0026] This invention, based on Traditional Chinese Medicine (TCM) theory, targets four major pathological factors: blood stasis, phlegm, toxins, and stagnation. Guided by the holistic concept of TCM, it proposes a treatment approach that combines attacking and tonifying, addressing both the root cause and the symptoms. The core formula uses *Bear Bile Wood* and *Viola yedoensis* to directly target the root cause and powerfully clear heat; *Earthworm* and *Smilax china* are used as supplementary ingredients to invigorate blood and unblock meridians; *Magnolia officinalis* and *Radix Raphanus sativus* are used as adjuvants to regulate qi and relieve stagnation; and *Atractylodes macrocephala* and *Dioscorea opposita* are added to strengthen the spleen and replenish qi, protecting the body's acquired constitution and ensuring that the body's vital energy remains strong despite the powerful attack on pathogens. The entire formula addresses the root cause of the disease. The current symptoms and root causes of the disease are addressed by using herbs such as Bletilla striata, cuttlebone, and Agrimonia pilosa to astringe and stop bleeding, and repair and regenerate tissue to treat the symptoms, as well as herbs that clear heat and detoxify, and strengthen the spleen and regulate qi to treat the root cause. The herbal composition of this invention has significant efficacy for different types of CAG in traditional Chinese medicine, such as liver and stomach qi stagnation syndrome, spleen and stomach weakness syndrome (spleen and stomach deficiency-cold syndrome), and liver and stomach heat stagnation syndrome. It can significantly alleviate the clinical symptoms of metaplastic atrophic gastritis, improve gastric mucosal atrophy and intestinal metaplasia, improve the quality of life of patients, and has high versatility and good safety.

[0027] This invention uses *Bruguiera gymnorhiza* as the principal ingredient, which clears heat and detoxifies, eliminates carbuncles and dissipates nodules, directly clearing "bacteria" and "inflammatory factors" in the stomach. Earthworm, *Bletilla striata*, *Atractylodes macrocephala*, *Dioscorea opposita*, and *Paeonia lactiflora* are used as assistant ingredients. Assistant ingredients assist the principal ingredient in strengthening the treatment of the main disease or symptom, or in treating concurrent diseases. Earthworm and *Bletilla striata* work together to invigorate blood circulation, remove blood stasis and dissipate nodules, astringe and stop bleeding, and repair damage. *Atractylodes macrocephala* and *Dioscorea opposita*, as assistant ingredients, are used to strengthen the spleen and stomach, replenish qi and blood, providing the fundamental driving force for the entire formula. To strengthen the body's vital energy, the combination of white peony root and atractylodes macrocephala can soothe the liver, relieve spasms, and alleviate pain. At the same time, it has the effect of nourishing blood and astringing yin, which can prevent other drugs in the formula from being too warm and dry or depleting yin and blood. It forms a "dynamic and static" combination with the blood-activating and collateral-clearing effect of the assistant drug earthworm. Agrimonia pilosa, Viola yedoensis, Smilax china, cuttlefish sheath, Magnolia officinalis, radish seed, and chicken gizzard are adjuvant drugs. The adjuvant drugs Magnolia officinalis and radish seed in this invention can also play the role of the guiding drug. By regulating the qi of the spleen and stomach in the middle jiao, they play a certain role in harmonizing and guiding.

[0028] Traditional Chinese medicine compound has multiple drug compositions, and plays a role in regulating multiple targets, multiple pathways and multiple systems in the treatment of diseases, effectively analyzing the material basis and mechanism of traditional Chinese medicine compound in treating diseases, which is an important task to promote the development of traditional Chinese medicine. In the present application, the blood entering components and metabolites of traditional Chinese medicine compound are comprehensively and accurately identified by using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC / Q-TOF-MS) technology, and then combined with network pharmacology, molecular docking, animal and molecular biology experiments, the active substances and mechanism of traditional Chinese medicine compound are determined. This research measure clearly and accurately describes the corresponding relationship between the components of the drug, the target and the disease, and reveals the characteristics of multi-component, multi-target and multi-system regulation of traditional Chinese medicine, which is consistent with the holistic concept of traditional Chinese medicine. In the present application, the CAG rat model is taken as the research object, and the UHPLC-Q-TOF-MS technology, molecular network technology, network pharmacology and ELISA technology are combined to explore the pharmacodynamic material basis and mechanism of the traditional Chinese medicine composition in treating CAG, and to provide theoretical and experimental basis for the clinical popularization and application of the traditional Chinese medicine composition in the present application.

[0029] The present application finds that terpenes, alkaloids and their derivatives and flavonoids are the main active substances of the present application through blood entering component identification; KEGG pathway enrichment analysis finds that the key target points of the present application in treating CAG are mainly enriched in cancer pathways, MAPK, PI3K-Akt and HIF-1 signaling pathways, and these pathways are closely related to inflammation and tumor biological network regulation targets; further, the present application can reduce the inflammation signaling pathway, oxidative stress signaling pathway and activate the apoptosis signaling pathway of gastric tissue; the present application can reduce the serum concentration of ALP, ALT and AST of patients with chronic atrophic gastritis, and has no hepatotoxicity and nephrotoxicity. BRIEF DESCRIPTION OF DRAWINGS

[0030] Figure 1 The total ion current diagram of the water extract under positive and negative ion modes of the present application;

[0031] Figure 2 The chemical composition distribution diagram of the present application;

[0032] Figure 3 The total ion current diagram of the blood plasma after entering the blood under positive and negative ion modes of the present application;

[0033] Figure 4 The blood entering component distribution diagram of the chemical substance of the present application;

[0034] Figure 5 The component-target network diagram of the present application;

[0035] Figure 6 A The intersection of the blood entering component target points and the CAG target points of the present application is the Wayne diagram;

[0036] Figure 7 GO function and KEGG pathway enrichment analysis of C and D core genes;

[0037] Figure 8 Rat gastric mucosa histopathology impact chart A blank control group B model group C the present application D positive drug group;

[0038] Figure 9 ROS detection chart of the present application;

[0039] Figure 10 mRNA expression level of GSH gene, mRNA expression level of SOD gene, mRNA expression level of MDA gene;

[0040] Figure 11 Tunnel staining chart of the present application;

[0041] Figure 12 WB chart of the present application for detecting cell apoptosis degree index;

[0042] Figure 13 Anti-inflammatory program index detection chart of the present application;

[0043] Figure 14 Liver toxicity index detection chart of the present application. DETAILED DESCRIPTION

[0044] In order to enable the personnel in the art to better understand the present application, the technical solutions in the embodiments of the present application will be described clearly and completely in conjunction with cases. Obviously, the described embodiments are only a part of the embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by the person in the art without creative labor should belong to the protection scope of the present application.

[0045] The experimental methods in the following examples are all conventional methods, and are carried out according to the techniques or conditions described in the literature in the art or according to the product instructions, unless otherwise specified. The materials, reagents and the like used in the following examples can be obtained from commercial channels, unless otherwise specified.

[0046] Example 1 Preparation of the composition of the present application

[0047] Formula: Xiongdanmu 16 parts, Dilong 16 parts, Baiji 20 parts, Baishu 20 parts, Baishao 15 parts, Shanyao 20 parts, Xianhecao 20 parts, Zihuadiding 15 parts, Guoshanlong 9 parts, Jineijin 9 parts, Hupu 9 parts, Haipiaoshe 15 parts, Laihu 20 parts

[0048] Bear bile wood has a special processing method: after the bear bile wood is peeled from the tree, the broken parts of the bark need to be sealed with a hot iron or other methods to seal the large amount of resin in the bear bile wood.

[0049] Weigh the above-mentioned proportions of Chinese medicinal herbs; dry the earthworm, cuttlebone, chicken gizzard lining, radish seed, and white peony root at 40℃; weigh 30g of honey, stir-fry the earthworm with wildflower honey until the white smoke dissipates and blue smoke appears, then remove from heat and set aside; remove the shell from the cuttlebone and crush it; stir-fry the chicken gizzard lining, radish seed, and white peony root separately until yellow and fragrant, then set aside; pass the earthworm, cuttlebone, chicken gizzard lining, radish seed, and white peony root through a 60-mesh sieve; grind the weighed Bletilla striata, yam, Agrimonia pilosa, Viola yedoensis, and Smilax china into coarse powder, add 500g of water and decoct, bring to a boil over high heat, then simmer over low heat for 1 hour, decoct twice, filter and combine the decoctions, then concentrate the filtrate into a thick paste over low heat; grind the weighed Atractylodes macrocephala and Magnolia officinalis into coarse powder, soak in 120g of alcohol for 1.5 hours, then reflux to extract 20g of the extract. Extract twice, combine with the thick paste extracted from Bletilla striata, Dioscorea opposita, Agrimonia pilosa, Viola yedoensis, and Smilax china, stir well, and then concentrate it into a thick paste over a low heat; finally, add the crushed earthworm, cuttlebone, chicken gizzard lining, radish seed, and white peony root to the combined thick paste.

[0050] Example 2: Preparation of the composition of the present invention

[0051] Formula: 20g of bear bile wood, 20g of earthworm, 30g of Bletilla striata, 30g of Atractylodes macrocephala, 20g of Paeonia lactiflora, 30g of Dioscorea opposita, 30g of Agrimonia pilosa, 20g of Viola yedoensis, 15g of Smilax china, 15g of chicken gizzard lining, 15g of Magnolia officinalis, 20g of cuttlefish skin, and 30g of radish seed.

[0052] Bear bile wood has a special processing method: after the bear bile wood is peeled from the tree, the broken parts of the bark need to be sealed with a hot iron or other methods to seal the large amount of resin in the bear bile wood.

[0053] Take the above weight parts of traditional Chinese medicine decoction pieces; dry diadromous earthworm, sea squirt, chicken gizzard sumac and white peony at 60°C; take 40g of honey, fry the earthworm with the honey until the white smoke disappears, and take it out when the smoke appears; break the sea squirt into pieces and set aside; fry the chicken gizzard, sumac and white peony separately until yellow and fragrant, and set aside; pass the earthworm, sea squirt, chicken gizzard, sumac and white peony through a 60-mesh sieve; grind the white yam, herbaceous peony, purple flower ground ivy and over mountain dragon into coarse powder, add 1000g of water and decoct, heat to boiling with strong fire, and decoct for 1.5h with gentle fire, decoct twice, filter and combine the decoction, and concentrate the filtrate into thick paste with gentle fire; grind the white atractylodes and magnolia officinalis into coarse powder, soak in 120g of alcohol for 2h, then extract for 30min with reflux, extract twice, combine with the thick paste extracted from white yam, herbaceous peony, purple flower ground ivy and over mountain dragon, stir well, and concentrate into thick paste with gentle fire; finally, add the ground earthworm, sea squirt, chicken gizzard, sumac and white peony into the combined thick paste.

[0054] Example 3 Preparation of the composition of the present application

[0055] Formulation: 10g of bear gall wood, 10g of earthworm, 10g of white yam, 10g of white atractylodes, 10g of white peony, 10g of herbaceous peony, 10g of purple flower ground ivy, 5g of over mountain dragon, 5g of chicken gizzard, 5g of magnolia officinalis, 10g of sea squirt, 10g of sumac.

[0056] The bear gall wood has a special processing method: after the bear gall wood is peeled off from the tree, the broken part of the bark needs to be sealed with a soldering iron or other means to seal a large amount of gum in the bear gall wood.

[0057] Take the above weight parts of traditional Chinese medicine decoction pieces; dry diadromous earthworm, sea squirt, chicken gizzard sumac and white peony at 40°C; take 40g of honey, fry the earthworm with the honey until the white smoke disappears, and take it out when the smoke appears; break the sea squirt into pieces and set aside; fry the chicken gizzard, sumac and white peony separately until yellow and fragrant, and set aside; pass the earthworm, sea squirt, chicken gizzard, sumac and white peony through a 60-mesh sieve; grind the white yam, herbaceous peony, purple flower ground ivy and over mountain dragon into coarse powder, add 1000g of water and decoct, heat to boiling with strong fire, and decoct for 1.5h with gentle fire, decoct twice, filter and combine the decoction, and concentrate the filtrate into thick paste with gentle fire; grind the white atractylodes and magnolia officinalis into coarse powder, soak in 120g of alcohol for 2h, then extract for 30min with reflux, extract twice, combine with the thick paste extracted from white yam, herbaceous peony, purple flower ground ivy and over mountain dragon, stir well, and concentrate into thick paste with gentle fire; finally, add the ground earthworm, sea squirt, chicken gizzard, sumac and white peony into the combined thick paste.

[0058] Example 4 Preparation of the composition of the present application

[0059] Ingredients: Xiongdanshu 20g, Dilor 20g, Baiji 30g, Baishu 30g, Baishao 20g, Shanyao 30g, Xianhecao 30g, Zihuadiding 20g, Guoshanlong 15g, Jineijin 15g, Houpu 15g, Haipiaoxie 20g, Laihu 30g

[0060] The above-mentioned raw materials are weighed according to the proportion, 8 times of water is added, decocted for 2 times, 30 minutes each time, filtered, and the filtrate is combined to obtain the medicinal liquid.

[0061] The beneficial effects of the present application are illustrated by the following test examples

[0062] Test Example 1 Clinical Research

[0063] Select patients with metaplastic chronic atrophic gastritis treated in Xiangxi Xiangyi Clinic of Hunan Province from December 2017 to December 2023, and randomly divide them into a treatment group and a control group, with 70 cases in the treatment group and 70 cases in the control group. There is no statistically significant difference (P>0.05) between the general data of the two groups, which is comparable. The treatment group uses the traditional Chinese medicine composition prepared by the present application Example 2, and the control group: selenium yeast tablets: produced by Mudanjiang Lingtai Pharmaceutical Co., Ltd., chewed after breakfast, 100 ug each time; the treatment course is 6 months.

[0064] (1) Inclusion and exclusion criteria

[0065] ① Patients diagnosed as CAG by endoscopy and pathological examination, without intraepithelial neoplasia, and pathological diagnosis standard refers to "Chinese Consensus on Chronic Gastritis"; patients with a history of gastric surgery, intraepithelial neoplasia, positive for Helicobacter pylori (except for negative for Helicobacter pylori after standard treatment) and severe diseases affecting the evaluation of curative effect (such as hematological diseases, liver diseases, heart diseases, kidney diseases, malignant tumors, etc.) are excluded; ② or negative for Helicobacter pylori after standard treatment; ③ age 18-75 years old, gender unrestricted; pregnant and lactating women are excluded; ④ subjects (or legal representatives) voluntarily agree and sign the informed consent form, and can comply with the research plan during the research period; patients who cannot correctly express subjective feelings or collaborators are excluded.

[0066] (2) Clinical symptom curative effect evaluation

[0067] The curative effect evaluation standard is formulated according to the "Guiding Principles for Clinical Research of New Traditional Chinese Medicine", and the integral observation table is used. The symptom curative effect judgment standard: the curative effect index is evaluated by using nimodipine method, curative effect index = [(pre-treatment integral - post-treatment integral) / pre-treatment integral] x 100%.

[0068] Cure: symptoms and signs disappear or basically disappear, efficacy index ≥ 95%; Marked effect: symptoms and signs are obviously improved, 70% ≤ efficacy index < 95%; Effective: symptoms and signs are improved, 30% ≤ efficacy index < 70%; Invalid: symptoms and signs are not obviously improved, or even aggravated, efficacy index < 30%; Total effective rate = (cured number + effective number + effective number) / total number × 100%.

[0069] (3) Gastric mucosa pathological evaluation

[0070] Pathological evaluation criteria: cure: intestinal epithelial metaplasia or atrophy returns to normal or disappears; Marked effect: intestinal epithelial metaplasia or atrophy is reduced by 2 degrees; Effective: intestinal epithelial metaplasia or atrophy is reduced by 1 degree; Invalid: does not reach the above effective standard, or worsens.

[0071] Gastric mucosa pathology score refers to the intuitive analog scoring method of the Chinese Consensus Opinion on Chronic Gastritis and the Sydney System, and atrophy, intestinal metaplasia, chronic inflammation and activity are divided into 4 grades of none, mild, moderate and severe, and are respectively given 0, 1, 2 and 3 points. Patients are respectively subjected to gastroscopy before treatment and within 3 months after treatment. The pathological sampling of the present study is performed under the high-definition gastroscopy of narrowband imaging. The present study mainly observes the changes of intestinal metaplasia at the antrum and angle of stomach, and the sampling position is the most serious place of intestinal metaplasia at the antrum and angle of stomach observed under endoscopy, and 1-2 pieces of biopsy can be taken as appropriate, and the antrum and angle of stomach are more serious. If other positions also appear serious lesions, pathological biopsy and regular observation are also required, but are not included in the score. The reexamination sampling position is the most serious position of intestinal metaplasia at the antrum and angle of stomach.

[0072] (4) Results

[0073] Table 1 Comparison of treatment effects of treatment group and control group

[0074] Group Total cases Cured Markedly effective Effective Ineffective Total effective rate Treatment group 70 15 30 20 5 92.9% Control group 70 0 15 25 30 57.14%

[0075] As can be seen from the above table, the traditional Chinese medicine composition of the present application has high effectiveness and can effectively treat atrophic gastritis.

[0076] Test Example 2 Typical Case

[0077] Case 1

[0078] Hu, female, 55 years old, because of "abdominal pain intermittent 7-8 years" to see a doctor, observation: patients feel bloating, stomach pain, with burning sensation, after eating, belching, sometimes back pain, dry stool, yellow, peeling, dark red, thin pulse. Auxiliary examination: gastroscopy: chronic gastritis with erosion, HP-. Gastroscopy pathology: (antrum) mild chronic atrophic gastritis with intestinal metaplasia. Pathogenesis: heat and water damage, wet heat resistance therapy: nourishing yin and generating fluid, clearing wet heat. To be treated with this party, daily a dose, morning and evening. 2 weeks after the patient's stomach pain, burning sensation disappeared, reduced, postprandial bloating, back pain relief, throat have sputum, dry stool, oral ulcer occasionally, thin yellow, peeling, dark, followed by taking this party. Take 2 months after the patient's stomach pain, burning sensation, significantly reduced, belching, defecation.

[0079] Case 2

[0080] Liu, female, 44 years old, because of "abdominal pain intermittent 3 years" to see a doctor, nearly 1 month, patients feel stomach pain more than before, observation: patients bloating, stomach pain, heartburn, noisy, during the tidal volume more block, dry mouth in the morning, dry stool, thin yellow, dark, pulse. Auxiliary examination: gastroscopy: chronic gastritis, HP-. Gastroscopy pathology: (antrum small) mild chronic atrophic gastritis with intestinal metaplasia. Pathogenesis: Qi and water damage, wet heat resistance. To be treated with this party, daily a dose, morning and evening. 2 weeks after the patient's stomach pain, slightly bloating, noisy, food, taste, mouth, dry, not bitter, normal stool, followed by taking this party. Take 2 months after the patient's stomach pain, bloating, heartburn, reduced, belching occasionally, dry stool slightly.

[0081] Case 3

[0082] Wang, male, 41 years old, because of "stuffy bloating intermittent 3 years" to see a doctor, observation: patients feel stomach burning sensation, stomach pain, postprandial abdominal distension, occasionally acid, noisy, dry mouth, stool shape, yellow, thin, tongue, pulse small slip. Past patients 3 years ago gastroscopy showed mild chronic atrophic gastritis with intestinal metaplasia. 2 years ago, gallbladder polyps cholecystectomy. 1 month ago, patients in Nanjing Drum Tower Hospital gastroscopy showed erosive gastritis, gastroscopy pathology showed (antrum small) mild chronic atrophic gastritis with intestinal metaplasia, lymphatic follicle formation 1. Abdominal B-ultrasound showed cholecystectomy, liver, spleen, bile duct, pancreas no obvious abnormalities. Colonoscopy showed no obvious abnormalities, pathogenesis: water and gas damage, wet heat resistance. To be treated with this party, daily a dose, morning and evening. Take 2 months after the patient's abdominal distension, stomach pain, postprandial, significantly reduced, dry mouth, noisy, acid, stool shape.

[0083] Case 4

[0084] Zhao, male, 40 years old. Suffering from stomach disease for 5 years, showing stomach pain and acid reflux, obvious reduction in diet, weak body, and tired appearance. Gastroscopy shows a few ulcer points with a large area of edema and congestion. Frequent use of western medicine and infusion treatment, but the curative effect is unstable. In October 2007, the decoction of the Chinese medicine composition was taken, and after 60 days of continuous taking, it was completely cured, and there was no recurrence so far.

[0085] Case 5

[0086] Wu, male, 46 years old. Suffering from stomach disease for more than 20 years, often belching, vomiting, acid reflux and pain. Gastroscopy shows that the stomach wall has erythema, multiple ulcers and erosion points. Take the pill of the Chinese medicine composition, the symptoms of nausea and vomiting disappear after 6 days, and improve significantly after 15 days. After 60 days of continuous taking, the gastroscopy recheck after 4 years shows that the symptoms of gastric erythema and erosion have completely disappeared.

[0087] The beneficial effects of the present application are illustrated by pharmacological experiments as follows

[0088] 1. Instruments and materials

[0089] Instruments: ultra-high liquid chromatography tandem Fourier transform mass spectrometry UPLC-Q-Exactive HF-X, Thermo Fisher Corporation; JXDC-20 type nitrogen purging instrument, Shanghai Jingxin Industrial Development Co., Ltd.; LNG-T88 type desktop rapid centrifugal concentration dryer, Jiangsu Taicang Huamei Biochemical Instrument Factory; Wonbio-96c type high-throughput tissue crusher, Shanghai Wanbai Biological Technology Co., Ltd.; SBL-10DT type ultrasonic cleaner, Ningbo Xinzhi Biological Technology Co., Ltd., 5430R type high-speed refrigerated centrifuge, Germany Eppendorf Company; MS105DU type electronic balance, Switzerland Mettler Company.

[0090] Materials: SPF male Wistar rats were purchased from the Animal Experiment Center of Anhui Medical University, and chromatographically pure methanol, acetonitrile, formic acid, propanol and water were purchased from Fisher Company, USA.

[0091] The water extract of the present application: take the formula: 20g of Ursi Cortex, 20g of Earthworm, 30g of Bletilla, 30g of Atractylodes, 20g of White Peony Root, 30g of Yam, 30g of St. John's Wort, 20g of Purple Flower, 15g of Over Mountain Dragon, 15g of Gizzard Gold, 15g of Magnolia Officinalis, 20g of Concha Arcae, 30g of Raphanus Sativus; After being pounded into powder, 1500 mL of water is added, soaked for 60 min, decocted to 500 mL, and the filtrate is taken; The residue is added with 500 mL of water, decocted to 200 mL, and the filtrate is taken, and the two filtrates are combined and concentrated to 150 mL of extract. The extract is rotary evaporated to obtain dry powder of the extract.

[0092] Animal model building and drug intervention:

[0093] SPF level male Wistar rats 90, after 1 week of adaptive feeding were randomly divided into normal control group and modeling group, 15 in the normal control group, 75 in the modeling group. The normal control group was given ordinary feed, free water. The modeling group was modeled by chemical method N-methyl-N-nitro-nitroso guanidine (MNNG), MNNG and irregular diet (fasting every other day) were given to the rats, and 100 g / mL MNNG solution was given by gavage every other day. After 10 weeks of modeling, 2 rats were randomly selected for gastric mucosa pathological observation. When the rats showed a decrease in the number of gastric glands, a thinning of the gastric mucosa, and inflammatory cell infiltration, the modeling was considered successful. The rats were then randomly divided into a model group, a high dose (13.5 g / kg) of the water extract of the application, a medium dose (4.5 g / kg) of the water extract of the application, and a low dose (1.5 g / kg) of the water extract of the application, and a Veseense control group, with 12-15 rats in each group. The drug treatment group was given corresponding drug gavage treatment, the normal control and model groups were given normal saline control treatment, and after 4 weeks of drug administration, the rats were humanely sacrificed, and the gastric tissue was fixed with polyformaldehyde or stored at -80°C for later use. All animal experiments were approved by the Animal Experiment Theory Committee of Anhui University of Chinese Medicine.

[0094] Example 5 Identification of blood components, core target identification, and target function research

[0095] 1. Plasma sample processing

[0096] At the end of the animal experiment, the rats were anesthetized by intraperitoneal injection of 1.5 g / kg urethane, and blood was taken from the abdominal aorta. The supernatant was obtained by centrifugation at 3500 r / min for 15 min. 100 μL of drug-containing plasma was added to 400 μL of extraction solution (acetonitrile:methanol=1:1) containing 0.02 mg / mL internal standard (L-2-chlorophenylalanine), vortexed for 30 s, and then low-temperature ultrasonic extraction was performed for 30 min (5°C, 40 KHz). The sample was placed at -20°C for 30 min. Centrifugation was performed at 4°C and 13000 g for 15 min, and the supernatant was removed. Nitrogen was used to dry the sample, and 100 μL of reconstitution solution (acetonitrile:water=1:1) was used for reconstitution. Low-temperature ultrasonic extraction was performed for 5 min (5°C, 40 KHz), and centrifugation was performed at 4°C and 13000 g for 10 min. The supernatant was analyzed by machine detection. The serum of the blank control group was treated in the same way.

[0097] 2. Compound identification and blood component analysis

[0098] The ion fragment information obtained by mass spectrometry of traditional Chinese medicine and plasma samples was introduced into the processing software Progenesis QI (Waters corporation, Milford, USA) for baseline filtering, peak identification, integration, retention time correction, peak alignment, and finally a data matrix of retention time, mass-to-charge ratio and peak intensity was obtained. At the same time, the MS and MS / MS mass spectrum information was matched with the public database HMDB (https: / / hmdb.ca / ) and Metlin, and the self-built library to obtain the annotation information of metabolites, and statistics. The original active substances of traditional Chinese medicine and the substances contained in the blood samples before and after the traditional Chinese medicine were analyzed, and the screening conditions of the blood components of traditional Chinese medicine were as follows: 1) the substances common to traditional Chinese medicine and serum after taking medicine but not in blank serum; 2) the substances contained in traditional Chinese medicine and the original peak value FC>2 in serum after taking medicine and blank serum. If the above two points are met, it is considered that the substance is a blood component.

[0099] 3. Prediction of blood components target, screening of core target and functional research

[0100] The blood components of the application screened were docked to the traditional Chinese medicine system pharmacology platform TCMSP and the drug information database Drug Bank to find the corresponding target genes. The blood components of traditional Chinese medicine and related target genes were introduced into Cytoscape 3.8.1 software to construct a traditional Chinese medicine blood active component-target network. The CAG disease related targets were obtained from DisGeNET database (www.disgenet.org) and GeneCards database (www.Genecards.org). Then the target corresponding to the blood components of the application and the potential target of CAG disease were intersected, and the obtained target was the potential target for treating CAG of the application. The protein-protein interaction (PPI) of the target related to the blood components was analyzed by String database, the interaction parameters were introduced into Cytoscape 3.8.1 for network visualization, and the network was topologically analyzed by CytoNCA software, and the median of betweenness centrality (BC), closeness centrality (CC) and degree centrality (DC) of the node was calculated to screen the core target. The key target was input into the DAVID database to export biological process (Biological process, BP), cellular component (Cellular component, CC), molecular function (Molecular function, MF) and KEGG pathway enrichment analysis of gene ontology function analysis.

[0101] 4. Experimental results 4.1 Identification of main chemical components of the application

[0102] The total ion fragment information obtained by mass spectrometry of the traditional Chinese medicine sample is compared with the Chemspider database, the chemicalbook database and the traditional Chinese medicine specific metabolite database (MJBIOTCM), and 563 chemical components are identified. The total ion chromatogram in positive and negative ion modes of the water extract and the component attribution are shown in Figure 1 A-B, and the chemical components of the present application are shown in Table 1. The top several components with the highest content are terpenes (25.54%), alkaloids and derivatives thereof (24.7%) and flavonoids (11.85%) Figure 2 A). The components with the largest amount are flavonoids (23.32%), terpenes (19.59%) and alkaloids and derivatives thereof (5.04%) Figure 2 B). In addition, the contents of sugars (6.26%), organic acids and derivatives thereof (4.26%) and amino acids and analogues thereof (3.32%) are also relatively rich Figure 2 A-B).

[0103] Table 1 Main chemical components of the present application

[0104]

[0105]

[0106]

[0107] 4.2 Identification of main blood components of the present application

[0108] The total ion fragment information obtained by mass spectrometry of the blood / target tissue is compared with the metabolite database and the local library to obtain the annotation information of the metabolites, and the statistics are obtained. The total ion chromatogram in positive and negative ion modes of the blood plasma is shown in Figure 3 A-B. The original active substances of the traditional Chinese medicine and the substances contained in the blood sample before and after the traditional Chinese medicine enters the blood are analyzed. The top several components with the highest content are alkaloids and derivatives thereof (15.32%), terpenes (5.58%) and flavonoids (2.29%) Figure 4 A). The components with the largest amount are terpenes (26.09%), alkaloids and derivatives thereof (15.22%) and flavonoids (13.04%) Figure 4 B). It is shown that the components with high content and multiple types of traditional Chinese medicine components entering the blood are also relatively more. Terpenes, alkaloids and derivatives thereof and flavonoids are the main active substances of the present application.

[0109] 4.3 Target screening of blood components of the present application based on network pharmacology

[0110] Among the 46 blood-entering components of the application, 41 components are predicted to have corresponding target points, and 259 target points are obtained in total through screening of the TCMSP database, Drug bank database and literature. Uniprot is used for gene name standardization, and a "component-target point" network thereof is constructed by using Cytoscape software Figure 5 According to the network topology analysis result, the Betweencentrality and Degree of the median are greater than 2, and the top few components with high node degree values are Genistein, Daidzein, Coclaurine and Stylopine. Among them, Genistein has 94 target points, Daidzein has 67 target points, Coclaurine has 41 target points, and Stylopine has 33 target points. The target points with high molecular degree values are PTGS2, PTGS1, ADRB2, CHRM1, HSP90, GABRA1 and the like. It is shown that these active components and molecules are important active substances and action target points for playing a biological role of the application.

[0111] 4.4 Target point function research of the application for treating CAG

[0112] The blood-entering component target points of the application and CAG-related genes are matched to obtain 91 target points (CAG) of the application for treating CAG Figure 6 The protein-protein interaction of the target points is analyzed by introducing a STRING database, and 23 key genes thereof are extracted by using Cytoscape software. It can be concluded that AKT1, TP53, TNF, EGFR, IL6 and BCL2 and other inflammation and immune regulation and cell proliferation and apoptosis-related target points are important target points of the application for treating CAG. GO function analysis shows that the key target points are mainly enriched in gene expression positive regulation, cell apoptosis process regulation, cell proliferation and other biological pathways; under the item of cell component, they are mainly enriched in nucleus, cytoplasm and mitochondria; under the item of molecular function, they are mainly protein, enzyme and DNA binding and the like. KEGG pathway enrichment analysis finds that the key target points of the application for treating CAG are mainly enriched in cancer pathway, MAPK, PI3K-Akt and HIF-1 signal pathway. These pathways are all closely related to inflammation and tumor biological network regulation target points Figure 7 C and Figure 7 D).

[0113] Example 5 Influence of the application on gastric mucosal lesions of CAG rats

[0114] 5.1 Experimental methods: Gastric tissue was fixed with paraformaldehyde for 24 hours, dehydrated with ethanol, cleared with xylene, embedded in paraffin, sectioned, stained with hematoxylin-eosin, and the pathological changes of gastric mucosa were observed under an optical microscope.

[0115] 5.2 Experimental Results: From Figure 8 The results showed that in the normal control group, the gastric mucosa and glandular structure of rats were intact, the muscular layer thickness was moderate, and there was no mucosal shedding or glandular atrophy. In the model group, the gastric mucosa of rats showed obvious inflammation, incomplete epithelial tissue structure, disordered muscular layer structure, and glandular atrophy. Compared with the model group, the number of gastric mucosal glands increased, inflammatory cell infiltration decreased, mucosal integrity improved, and muscular layer structural disorder lessened in the present invention group and the positive drug group. This indicates that the pharmaceutical composition of the present invention can significantly promote the repair of the gastric mucosa in rats.

[0116] Example 6 Oxidative Stress Examination

[0117] 1.1 Experimental Methods: Pre-coated cells were treated with your drug or stimulant for a predetermined time; DCFH-DA fluorescent probe was added and incubated in the dark for 20-30 minutes to allow the probe to enter the cells; after washing away excess probe with PBS, the green fluorescence intensity of the cells was immediately detected by fluorescence microscopy, microplate reader or flow cytometry; the stronger the green fluorescence, the higher the level of reactive oxygen species (ROS) produced in the cells.

[0118] 1.2 Experimental Results: ROS, GSH, SOD, and MDA are indicators related to oxidative stress. Figure 9 The results show that the more red light there is, the higher the expression of ROS, indicating that traditional Chinese medicine significantly improved the level of oxidative stress. Furthermore, PCR showed that... Figure 10 (In the figure, C is the blank control group, M is the model group, DPCT is the present invention, and AC is the positive drug group.) It can be seen that the expression of GSH, SOD, and MDA is also reduced under the action of traditional Chinese medicine.

[0119] Example 7 Apoptosis degree examination

[0120] 7.1 Experimental methods: After inducing apoptosis by treating cells with drugs or stimulants, the cells were digested and collected to prepare a single-cell suspension; Annexin V (labeling early apoptosis) and PI (labeling late apoptosis / necrosis) dyes were added and incubated at room temperature in the dark for about 15 minutes; the stained cells were immediately detected by flow cytometry to analyze the fluorescence signal of the cells.

[0121] 7.2 Experimental Results: As shown in the figure, the more green there is, the stronger the apoptosis. Figure 11 The study showed that traditional Chinese medicine reduced apoptosis in gastric tissue. BAX, CAPSASE3, and BCL2 are apoptosis-related markers.Figure 12 WB showed that under the action of the traditional Chinese medicine, the transcription and translation of BAX and CAPSASE3 were reduced, and the transcription and translation of BCL2 were increased, indicating that the traditional Chinese medicine alleviated apoptosis.

[0122] Example 8 Inflammation degree detection

[0123] 8.1 Experimental method: serum of each group of rats was taken; protein sample preparation; gel electrophoresis; membrane transfer; blocking; first antibody incubation; second antibody incubation; signal detection.

[0124] 8.2 Experimental results: IL-6, TNF-a and IL-1b are inflammatory factors, from Figure 13 It was found that the traditional Chinese medicine compound in the application could prevent inflammatory response at the gene and protein levels.

[0125] Example 9 Hepatotoxicity

[0126] Using a liver function detection kit, from Figure 14 It was found that the application could reduce the serum concentrations of ALP, ALT and AST of patients with chronic atrophic gastritis.

Claims

1. A traditional Chinese medicine composition for treating chronic atrophic gastritis, characterized in that, The ingredients include the following parts by weight: 10-20 parts of bear bile wood, 10-20 parts of earthworm, 10-30 parts of Bletilla striata, 10-30 parts of Atractylodes macrocephala, 10-20 parts of white peony root, 10-30 parts of yam, 10-30 parts of Agrimonia pilosa, 10-20 parts of Viola yedoensis, 5-15 parts of Smilax china, 5-15 parts of chicken gizzard lining, 5-15 parts of Magnolia officinalis, 10-20 parts of cuttlefish sheath, and 10-30 parts of radish seed.

2. The traditional Chinese medicine composition according to claim 1, characterized in that: The ingredients include the following parts by weight: 16 parts of bear bile wood, 16 parts of earthworm, 20 parts of Bletilla striata, 20 parts of Atractylodes macrocephala, 15 parts of white peony root, 20 parts of yam, 20 parts of Agrimonia pilosa, 15 parts of Viola yedoensis, 9 parts of Smilax china, 9 parts of chicken gizzard lining, 9 parts of Magnolia officinalis, 15 parts of cuttlefish skin, and 20 parts of radish seed.

3. The traditional Chinese medicine composition as described in claim 1 or 2, characterized in that: The composition also includes 20-40 parts or 30 parts of honey.

4. The traditional Chinese medicine composition according to claim 3, characterized in that: The bear bile wood is rich in resin, and the broken parts of the bear bile wood need to be sealed with resin using a soldering iron.

5. The composition according to any one of claims 1-4, characterized in that: The composition is a formulation prepared by adding pharmaceutically acceptable excipients to a powdered active pharmaceutical ingredient or an aqueous or organic solvent extract of the active pharmaceutical ingredient.

6. The composition according to claim 5, characterized in that: The preparation is a solution, granules, pills, ointment, or capsule.

7. Use of the traditional Chinese medicine composition according to any one of claims 1-6 in the preparation of a medicament for treating chronic atrophic gastritis.

8. The use as described in claim 7, characterized in that: The traditional Chinese medicine composition can reduce gastric tissue inflammation, oxidative stress, and activate apoptosis signaling pathways.

9. The use as described in claim 7, characterized in that: The traditional Chinese medicine composition reduces gastric tissue inflammation, oxidative stress, and activates apoptosis signaling pathways by acting on MAPK, PI3K-Akt, and HIF-1 signaling, respectively.

10. The use as described in claim 7, characterized in that: The traditional Chinese medicine composition can reduce serum concentrations of ALP, ALT, and AST.

Images