CN112206269A

CN112206269A - A Chinese medicinal composition for treating chronic benign gastropathy and esophageal diseases

Info

Publication number: CN112206269A
Application number: CN202011256880.0A
Authority: CN
Inventors: 孙喜灵; 张涛; 范庆波; 林剑; 高永林; 于帆; 林惠宇
Original assignee: Yantai University
Current assignee: Yantai University
Priority date: 2020-11-11
Filing date: 2020-11-11
Publication date: 2021-01-12

Abstract

The invention discloses a traditional Chinese medicine composition for treating chronic benign gastropathy and esophageal diseases, which is characterized by being prepared from the following raw medicinal materials, by weight, 8-12 parts of codonopsis pilosula, 8-12 parts of fried bighead atractylodes rhizome, 8-12 parts of poria cocos, 4-8 parts of fingered citron, 4-8 parts of immature bitter orange, 4-8 parts of hovenia dulcis thumb, 4-8 parts of sea buckthorn, 4-8 parts of perilla leaf, 4-8 parts of dark plum, 4-8 parts of seville orange flower, 4-8 parts of ginger processed pinellia tuber and 8-12 parts of liquorice; by selecting the specific traditional Chinese medicine components, the stomach illness and the esophagus disease caused by insufficient secretion of gastric acid and digestive enzyme can be effectively treated.

Description

A Chinese medicinal composition for treating chronic benign gastropathy and esophageal diseases

Technical Field

The invention relates to the field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition for treating chronic benign gastropathy and esophageal diseases.

Background

In recent years, the incidence of gastroenteropathy has been on the rise due to the increase of living pressure and the acceleration of pace, and the change of dietary structure and the bad dietary habits, such as overeating, overeating with fat and sweet taste, alcoholism and the like. The world health organization has statistically found that the incidence of gastrointestinal disorders is as high as 80% and is growing at a rate of around 20.0% per year. Gastrointestinal diseases do not directly threaten life, but bring great pain to patients, seriously affect the health and life quality of people, and if the gastrointestinal diseases are not treated in time, the condition of the patients is gradually worsened, even cancer is caused, and the life is threatened. Although the number of the Chinese patent medicines on the market is small, some Chinese patent medicines have poor curative effect, some Chinese patent medicines have higher price and some Chinese patent medicines are not easy to accept by patients with dosage forms, so the existing medicines on the market cannot meet the market requirement.

Clinical data analysis and research show that the rapid development of society and science and technology makes the working environment, rhythm, dietary structure, habit and the like of people greatly changed, and the basic nature of the gastropathy attack is greatly changed. The stomach diseases caused by excessive secretion of gastric acid and digestive enzyme are gradually reduced; stomach diseases caused by insufficient secretion of gastric acid and digestive enzymes are gradually increasing. The traditional study and application of the stomach disease medicament mainly take the medicaments for neutralizing gastric acid and inhibiting gastric acid secretion; in contrast, there has been a lack of research and use of drugs that increase gastric acid and promote gastric acid secretion. Therefore, the research on the drugs for treating the gastropathy based on the hyposecretion of gastric acid and digestive enzyme has very important theoretical and clinical values.

The formula and the preparation are guided by the principles of syndrome differentiation and disease diagnosis and treatment of traditional Chinese medicine, through clinical application big data for more than 20 years, and are combined with four methods of acid inhibition, gastric mucosa protection, gastric motility promotion and sterilization of modern medicine for treating gastropathy, and the formula method of acid promotion, gastric mucosa protection, gastric motility promotion and sterilization is provided for chronic gastritis with gastric acid and digestive enzyme secretion deficiency, and finally the effective treatment preparation for diseases, syndromes and symptoms is prepared by compatibility and combination. However, how to make the inventive formula exert higher potency and effect according to the physiological and pathological characteristics of the stomach becomes a key problem to be researched. The traditional Chinese medicine is combed by the traditional processing theory, and the modern microbial fermentation technology is introduced, so that the traditional Chinese medicine achieves the aims of attenuation and synergism, and the method is an effective way and an effective method for improving the clinical curative effect of the formula.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a traditional Chinese medicine composition for treating chronic benign gastropathy and esophageal diseases, and the composition can be used for treating the gastropathy and the esophageal diseases caused by insufficient secretion of gastric acid and digestive enzymes by tonifying spleen and stomach, regulating qi, resolving food stagnation and promoting fluid production to stop flow.

In order to solve the technical problem, the invention adopts the following technical scheme:

a traditional Chinese medicine composition for treating chronic benign gastropathy and esophageal diseases is characterized by being prepared from the following raw medicinal materials in parts by weight: 8-12 parts of codonopsis pilosula, 8-12 parts of fried bighead atractylodes rhizome, 8-12 parts of poria cocos, 4-8 parts of fingered citron, 4-8 parts of immature bitter orange, 4-8 parts of hovenia dulcis thumb, 4-8 parts of sea buckthorn, 4-8 parts of perilla leaf, 4-8 parts of dark plum, 4-8 parts of seville orange flower, 4-8 parts of ginger processed pinellia and 8-12 parts of liquorice.

The raw material medicinal materials are preferably in proportion: 9-11 parts of codonopsis pilosula, 9-11 parts of fried bighead atractylodes rhizome, 9-11 parts of poria cocos, 5-7 parts of fingered citron, 5-7 parts of immature bitter orange, 5-7 parts of hovenia dulcis thumb, 5-7 parts of sea buckthorn, 5-7 parts of perilla leaf, 5-7 parts of dark plum, 5-7 parts of seville orange flower, 5-7 parts of ginger processed pinellia tuber and 9-11 parts of liquorice.

The raw medicinal materials are further preferably proportioned as follows: 10 parts of codonopsis pilosula, 10 parts of fried bighead atractylodes rhizome, 10 parts of poria cocos, 6 parts of fingered citron, 6 parts of immature bitter orange, 6 parts of hovenia dulcis thunb, 6 parts of sea buckthorn, 6 parts of perilla leaf, 6 parts of dark plum, 6 parts of seville orange flower, 6 parts of pinellia ternate and 10 parts of liquorice.

The present invention relates to the following medicine materials:

codonopsis pilosula: has sweet and neutral nature, and has effects of invigorating spleen, replenishing qi, invigorating spleen, and benefiting lung, and can be used for treating spleen and lung weakness, short breath, cardiopalmus, anorexia, loose stool, asthmatic cough, internal heat, and diabetes.

White atractylodes rhizome: bitter and sweet in nature, warm in nature, entering spleen and stomach channels, has effects of invigorating spleen and qi, eliminating dampness and promoting diuresis, and arresting sweating, and can be used for treating spleen deficiency, anorexia, abdominal distention, diarrhea, phlegm and fluid retention, palpitation, edema, and spontaneous perspiration.

Tuckahoe, poria cocos: has sweet and bland nature and taste, and has effects of invigorating spleen, inducing diuresis, eliminating dampness, and calming heart, and can be used for treating edema, oliguria, phlegm retention, dizziness, palpitation, spleen deficiency, anorexia, loose stool, diarrhea, uneasiness, palpitation, and insomnia.

Fingered citron: the medicine has pungent, bitter, sour and warm natured properties, enters liver, spleen and lung channels, has the effects of soothing liver, regulating qi, harmonizing stomach and relieving pain, and is used for treating chest and hypochondrium distending pain, epigastric fullness, anorexia and vomiting caused by liver and stomach qi stagnation.

Citron: pungent, bitter and sour in nature, entering liver, spleen and lung meridian, with effects of soothing liver, regulating qi, relieving epigastric distention, and eliminating phlegm, and can be used for treating chest and hypochondrium distending pain, abdominal fullness, emesis, belching, excessive phlegm and cough due to qi stagnation of liver and stomach.

Immature bitter orange: bitter and octanoic acid in property, warm in property, entering spleen and stomach channels, has the effects of breaking qi, removing food retention, resolving phlegm and dispersing mass, and is used for treating mass and fullness, distending pain, diarrhea and diarrhea, constipation obstruction and phlegm stagnation and qi obstruction in chest and chest stagnation.

Raisin tree seed: has sweet and mild property and taste, and has effects of clearing heat, promoting urination, relieving cough, relieving restlessness, and relieving alcoholism, and can be used for treating fever polydipsia, singultus, emesis, dysuresia and alcoholism.

Sea-buckthorn: the nature and taste are warm and sour, and the medicine has the functions of invigorating spleen, stomach, lung and heart channels, expelling phlegm and arresting coughing, promoting digestion and resolving stagnation, and promoting blood circulation and removing blood stasis.

And (3) perilla leaves: has pungent and warm nature and flavor, and has effects of entering lung and spleen channels, relieving exterior syndrome, dispelling cold, activating qi-flowing, and regulating stomach function, and can be used for treating cough, nausea, vomiting of pregnancy, fish and crab poisoning.

Dark plum: the medicine has sour and astringent taste, enters liver, spleen, lung and large intestine channels, has the functions of astringing lung and intestine and promoting the production of body fluid, and is used for treating chronic cough, chronic dysentery, intestine dryness, deficiency heat, thirst and abdominal pain.

Substitute flowers: has pungent, sweet, slightly bitter and mild nature, and has the functions of regulating qi, relieving chest stuffiness, regulating stomach function, and relieving vomit, and can be used for treating chest distress, abdominal distention and pain, anorexia, nausea and emesis.

Ginger processed pinellia: with pungent and warm nature, spleen, stomach and lung meridians entered, it has effects of eliminating dampness and phlegm, lowering adverse qi, relieving vomit, relieving stuffiness and resolving hard mass, and can be used for treating excessive phlegm, cough and asthma, phlegm retention, dizziness, phlegm, headache, emesis, regurgitation, chest and abdominal distention, and globus hystericus.

Licorice root: has sweet and mild nature, and has effects of invigorating spleen and replenishing qi, clearing heat and detoxicating, eliminating phlegm and relieving cough, relieving spasm and pain, and harmonizing the medicines, and can be used for treating asthenia, cardiopalmus and short breath, cough with excessive phlegm, and spasm and pain of abdomen and limbs due to weakness of spleen and stomach, and relieving drug toxicity intensity.

The compatibility principle of the medicinal materials in the invention is as follows:

the codonopsis pilosula is a monarch drug, is used for tonifying middle-jiao and Qi, invigorating spleen and benefiting lung, and plays a main treatment role aiming at the principal symptoms. The atractylodes macrocephala koidz and the tuckahoe are adjuvant drugs, have the effects of strengthening the spleen and inducing diuresis together and have the effect of assisting to strengthen the spleen of the codonopsis pilosula. The fingered citron, the immature bitter orange, the hovenia dulcis thunb, the sea backthern, the perilla leaf, the dark plum, the seville orange flower and the pinellia ternate are all adjuvant drugs, and have the effects of soothing the liver, regulating qi, eliminating phlegm, relieving cough, harmonizing the stomach, removing stagnation and the like so as to relieve secondary concurrent syndrome and treat various symptoms of stomach diseases. The liquorice is used as a guiding drug to coordinate the effects of the various drugs and relieve the toxicity of the drugs in the prescription, so that the liquorice can combine with the drugs to eliminate pathogenic factors.

The specific dialectical treatment of the medicinal materials in the invention is as follows:

the clinical symptoms of chronic benign gastropathy and esophagus are presented, and the symptoms are single, two, three or a plurality of symptoms. The application of fingered citron, immature bitter orange, perilla leaf and seville orange flower can regulate qi, harmonize stomach and relieve pain; the stir-fried bighead atractylodes rhizome, immature bitter orange and ginger processed pinellia tuber are used for regulating qi, harmonizing stomach and removing stuffiness; the codonopsis pilosula, the fried bighead atractylodes rhizome and the poria cocos can tonify the spleen and the stomach and promote transportation and transformation; singultus, vomit qi, nausea and vomiting, or vomiting food and excessive mucus, which are caused by stomach qi failing to descend and stomach qi going upwards, ginger processed pinellia tuber and perilla leaf can harmonize stomach, descend adverse qi, arrest vomiting and stop vomiting; the stomach sound or the abdomen sound is caused by water-dampness in the middle warmer, and the fried rhizoma atractylodis macrocephalae, poria cocos, perilla leaves and seville orange flowers are used for tonifying spleen and resolving dampness and promoting the transportation and transformation of water-dampness; the bitter orange, the sea buckthorn, the fingered citron, the citron and the dark plum can clear heat and eliminate stagnation, harmonize stomach and eliminate fire; the stir-fried white atractylodes rhizome, immature bitter orange, finger citron, citron fruit and seville orange flower are used for regulating qi to alleviate pain when the spleen qi stagnation or the large intestine qi stagnation is accompanied with abdominal distension and stuffiness and pain; the bitter orange, the fingered citron, the hovenia dulcis thumb and the seville orange flower are used for regulating the flow of qi, relieving chest stuffiness and clearing heat, and are caused by stomach qi and stomach fire blocking in a gastric cavity and an upper adverse chest; the application of fingered citron, hovenia dulcis thunb, sea buckthorn, perilla leaf, dark plum and seville orange flower can regulate qi, relieve sore throat, clear heat and reduce phlegm; the combination of Dang Shen and Stir-baked Bai Zhu can tonify qi and check sweating, which is caused by weakness of spleen and stomach and failure of qi to check sweating. The formula of the invention can be analyzed from the dialectical treatment process of clinical application, and has obvious advantages for treating various stomach diseases caused by the insufficiency of gastric acid and digestive enzyme and the insufficiency of gastrointestinal motility.

The invention has the positive effects that:

the traditional Chinese medicine formula disclosed by the invention has the main effects of tonifying spleen and stomach, regulating qi, removing food retention and promoting fluid production to reverse flow, and has a reliable and effective effect on treating various stomach diseases and various symptoms caused by insufficiency of gastric acid and digestive enzyme. The symptoms of epigastric fullness or pain, epigastric fullness or hardness, anorexia, poor appetite, emaciation, hiccup, vomit gas or nausea, vomiting food, excessive mucus vomiting, gastric borborygmus or borygmus, epigastric upset or heartburn, fever discomfort, abdominal fullness and pain, chest oppression, burning sensation, pain, throat discomfort, blockage, phlegm and heat sensation, or dry mouth, bitter mouth, sticky mouth or fire, or lack of strength and sweating are all in the treatment range of the invention.

After treatment of one course of treatment, 81 percent of patients have obvious clinical relief of various symptoms; after three to four treatments, the clinical symptom relieving rate is 96 percent. Patients with atrophic gastritis need to continue recovery treatment of pathological changes for one month after the clinical symptoms have been reduced or disappeared. After the treatment course is finished, the result is checked again by the gastroscope, and the gastroscope and pathological results of 76% of patients are obviously improved or recovered to be normal.

Drawings

FIG. 1 is a liquid chromatogram of an unfermented drug solution in a liquid phase analysis according to the present invention;

FIG. 2 is a liquid chromatogram of the fermented liquid medicine in the liquid phase analysis of the present invention;

FIG. 3 is a SEC-HPLC chromatogram of an unfermented drug solution in a capillary electrophoresis analysis in accordance with the present invention;

FIG. 4 is a SEC-HPLC chromatogram of a post-fermentation drug solution in a capillary electrophoresis analysis in accordance with the present invention;

FIG. 5 is a graph of HE staining of pathological sections of stomach tissue of a blank group of animals in a rat model experiment of Chronic Atrophic Gastritis (CAG);

FIG. 6 is a graph of HE staining of animal gastric histopathological section of experimental model group of rat Chronic Atrophic Gastritis (CAG);

FIG. 7 is a graph of HE staining of pathological sections of stomach tissue of high dose animals in experimental sample groups of rat Chronic Atrophic Gastritis (CAG) model;

FIG. 8 is a photograph showing HE staining of pathological sections of stomach tissue of animals in a dose group in a rat Chronic Atrophic Gastritis (CAG) model experimental sample group.

Detailed Description

For a better understanding and appreciation of the invention, the invention will be further illustrated with reference to the accompanying drawings and specific examples.

Example 1

The preparation is prepared from the following raw medicinal materials in parts by weight: 10 parts of codonopsis pilosula, 10 parts of fried bighead atractylodes rhizome, 10 parts of poria cocos, 6 parts of fingered citron, 6 parts of immature bitter orange, 6 parts of hovenia dulcis thunb, 6 parts of sea buckthorn, 6 parts of perilla leaf, 6 parts of dark plum, 6 parts of seville orange flower, 6 parts of pinellia ternate and 10 parts of liquorice.

The traditional Chinese medicine composition is prepared according to the following steps:

preparing a culture medium: 94g of traditional Chinese medicine composition for treating chronic benign gastropathy and esophageal diseases is prepared according to the proportion, the traditional Chinese medicine composition is respectively filled into 4 triangular bottles with 300mL, and 33.4 mL of sterile water, 2.25 g of bran and 2.25 g of sweet wormwood are added into each triangular bottle;

(II) sterilization: placing the triangular flask into a sterilizing pot, and sterilizing at 121 deg.C under 0.12 MPa for 20 min;

(III) inoculation: taking out the triangular bottles from the sterilizing pot after cooling, and respectively adding 2.8 × 106-4.2 × 106 dry materials of food and medicine aspergillus CICC 41649;

(IV) fermenting: placing the triangular flask in an incubator at 30 ℃ for standing culture for 4.5 days, immediately buckling the flask when green mycelia grow out of the triangular flask, and then buckling the flask once every 12 h for culture for 4.5 days until fermentation is completed;

(V) decocting:

(1) first-time decoction: placing the fermented traditional Chinese medicine composition in a pressure cooker, adding distilled water with the amount of 5 times of the traditional Chinese medicine composition, soaking for 30 minutes, boiling with strong fire, then decocting with slow fire for 15 minutes, filtering to obtain medicinal residue, and collecting medicinal liquid;

(2) decocting for the second time: directly adding distilled water with 5 times of the amount of the residues into the residues filtered after the first decoction, boiling with strong fire, adjusting the fire to boil for 15 minutes, filtering to obtain residues, and collecting liquid medicine;

(3) mixing: mixing the liquid medicine obtained by the first decoction and the liquid medicine obtained by the second decoction to obtain a final liquid medicine product;

(VI) preparing medicine granules: further concentrating the medicinal liquid product, adding matrix required by pill preparation according to a proportion, mixing, and processing into medicinal granules.

Example 2

In this example, the raw material herbs and the ratio are the same as those in example 1, and when preparing the traditional Chinese medicine composition, the fermentation system enlarged to 30L is used for small-scale industrial preparation based on example 1, and the steps are as follows:

preparing a culture medium: 4.32 kg of a traditional Chinese medicine composition for treating chronic benign gastropathy and esophageal diseases is prepared according to the same proportion as that of the embodiment 1, 2.6L of sterile water, 405 g of bran and 405 g of sweet wormwood are added, and the prepared culture medium is subpackaged in 7 sterile feeders;

(II) sterilization: placing the aseptic feeder into a sterilizing pot, and sterilizing at 121 deg.C under 0.12 MPa for 20 min;

(III) inoculation: loading the sterilized Chinese medicinal composition into an empty fermentation system by using an aseptic feeder, inoculating 12% Aspergillus CICC41649 bacterial suspension for food and medicine by flame inoculation, and stirring at 5r/min for 40 min;

the flame inoculation method specifically comprises the following steps: dipping alcohol in the inoculating ring, sleeving the inoculating cap on the inoculating ring, slightly unscrewing the inoculating cap, and quickly adjusting the rotameter to 0.1 m³H; igniting a fire ring, slightly unscrewing an inoculation cap, placing the inoculation cap above flame, slightly pulling off the cotton plug beside the flame ring, then burning the bottle mouth of the triangular flask filled with the prepared bacterial suspension on the flame for 4 seconds, and quickly pouring the bacterial suspension and sterile water into a fermentation system; the mouth of the triangular flask does not leave the flame range, and after the inoculation of the seed bottle is completed, the inoculation cap is put on the flame to burn for 4 seconds, the flame ring is quickly screwed down, the fire ring is removed, and the fire ring is extinguished by using a wet towel.

(IV) fermenting: closing the fermentation tank after inoculation, and standing and culturing for 8 days in the fermentation tank at the temperature of 30 ℃ for 192 h; in the fermentation process, sterile air is introduced in stages, the ventilation volume of the first 12 h is 1.7L/min, the ventilation volume of the first 12-48 h is 3.3L/min, the ventilation volume of the first 48-96 h is 6.6L/min, and the ventilation volume of the first 96-192 h is 3.3L/min; in the fermentation process, stirring is carried out in stages, stirring is not carried out within 0-48 h, stirring is carried out once every 12 h within 48-192 h at the rotating speed of 5r/min, and the stirring time is 40 min each time.

(V) decocting:

And (4) conclusion: example 1 the traditional Chinese medicine formula was fermented in a 300mL triangular flask, and the liquid chromatogram peak area of the water-soluble components after fermentation was reduced by 94.23%; in example 2, the process is expanded to a fermentation system of 30L, and the liquid chromatographic peak area of the water-soluble components is reduced by 90.38% after fermentation; example 2 the liquid chromatographic peak area of the water soluble components after fermentation was reduced by 3.85% compared to example 1, indicating that the experimental process of example 1 gave a product superior to example 2, but the difference was tolerable as long as it was within the range of 10%. Therefore, the scale-up process of example 2 is basically satisfactory for industrial implementation.

Experimental example 1: experiment for promoting gastrointestinal peristalsis of mice

1.1 Experimental methods

1.1.1 atropine modeling experiment

50 male Kunming mice with the age of 3 weeks weigh 18-20 g. After 1 week of adaptive feeding, mice were randomly divided into normal control group, model group (atropine group), sample high, medium and low dose groups (sample + atropine). Purified water is used as a solvent. The sample concentrated solution (equivalent to 20 g/kg of crude drug, the same applies hereinafter), 10 g/kg (equivalent to 10 g/kg of crude drug, the same applies hereinafter), and 5 g/kg (equivalent to 5 g/kg of crude drug, the same applies hereinafter) were administered to the high, medium, and low dose fractions of the sample by intragastric administration, respectively. The gavage is continued for 15 days. And respectively gavage pure water with equal volume in the normal control group and the model group.

1.1.2L-arginine Molding experiment

50 male Kunming mice with the age of 3 weeks weigh 18-20 g. After 1 week of adaptive feeding, mice were randomly divided into normal control group, model group (L-arginine group), sample high, medium and low dose groups (sample + L-arginine). Purified water is used as a solvent. The high, medium and low dosage fractions of the sample were administered by intragastric administration to 40 g/kg (equivalent to 40 g/kg of crude drug, the same below), 20 g/kg (equivalent to 20 g/kg of crude drug, the same below), and 10 g/kg (equivalent to 10 g/kg of crude drug, the same below), respectively, of the sample concentrate. The gavage is continued for 15 days. The normal control group and the model group are respectively given purified water with equal volume. Starting on the 10 th day of gavage, mice in the model group and the sample group were simultaneously administered L-arginine (2 mg/kg) solution for gavage 1 time a day for 5 days.

1.1.3 determination of Small intestine Propulsion Rate

After the mice in each group are taken for 12 hours for the last time, the mice are fasted for 24 hours, water is freely drunk, and the weight of the mice in each group is accurately weighed before the experiment. Atropine molding experimental animals are given 0.5 mg/kg of atropine for each animal. Meanwhile, each group of mice is subjected to intragastric administration by ink and purified water. After 20 minutes, the mouse was sacrificed by cervical spondylolysis, gastrointestinal tissues were immediately separated by laparotomy, intestinal tissues were laid flat and kept in a tension-free state, the length of the small intestine was measured, and the distance from the pylorus to the leading edge of the ink was measured as the advancing distance of the ink in the small intestine.

The animals in the L-arginine molding experimental group are subjected to intragastric administration by ink and purified water. The mouse is killed by cervical spondylolysis after 20 minutes, gastrointestinal tissues are immediately dissected and separated, intestinal tissues are laid flat and kept in a tension-free state, the length of the small intestine is measured, and the distance from the pylorus to the front edge of the ink is measured as the advancing distance of the ink in the small intestine.

Calculating the ink propulsion rate by using a formula:

ink propulsion rate (%) = distance (cm) ink is propelled in small intestine/total length (cm) × 100%.

1.2 results of the experiment

1.2.1 atropine modeling experiment results

Compared with a normal control group, the intestinal propulsion rate of the mice in the model group is obviously reduced (P is less than 0.01) after the atropine is injected. Compared with the model group, the small intestine propulsion rate of mice given the high and medium dose groups (40 g crude drug/kg and 20g crude drug/kg) of the samples after the injection of the atropine is obviously enhanced compared with the atropine group (P is less than 0.01 and P is less than 0.05). Among them, the rate of intestinal transit was close to that of normal control mice in the high dose group (40 g crude drug/kg) mice (Table 1).

TABLE 1 Effect of samples on the rate of intestinal propulsion of mice by atropine

Group of	Number of animals	Small intestine propulsion rate (%)
			Normal control group	10	70.2±5.7
Model set (atropine set)	10	48.9±6.8^##
			Sample high dose group (40 g crude drug/kg)	10	68.3±5.4^**
Medium sample dose group (20 g crude drug/kg)	10	58.4±8.3^*
			Sample Low dose group (10 crude drug g/kg)	10	47.8±7.0

^##P is less than 0.01 compared with the normal control group; p < 0.05, P < 0.01 compared to model group.

1.2.2L-arginine modeling test results

Compared with the normal control group, the intestinal propulsion rate of the model group mice is obviously reduced compared with the propulsion rate of the normal control group mice (P is less than 0.01). Compared with the model group, the mice given with high and medium dose (40 g crude drug/kg and 20g crude drug/kg) samples have obviously enhanced intestinal propulsion rate (P < 0.01 and P < 0.05). See table 2 for details.

TABLE 2 influence of samples on the rate of intestinal transit of mice by L-arginine

Group of	Number of animals	Small intestine propulsion rate (%)
			Normal control group	10	65.4±7.1
Model set (L-arginine set)	10	41.3±5.8^##
			Sample high dose group (40 g crude drug/kg)	10	59.8±4.4^**
Sample middle dose group (20 g crude drug/k)g）	10	52.8±8.1^*
			Sample Low dose group (10 g crude drug/kg)	10	40.5±6.6

^##Comparing P with normal control group to be less than 0.01;^*，**compared with the model group, P is less than 0.05 and P is less than 0.01.

1.3 conclusion

The above results show that: the traditional Chinese medicine composition prepared by the invention has the effect of promoting gastrointestinal motility of mice. The lowest effective dose is 20g crude drug/kg, and the dose is converted into human clinical equivalent dose of 100 g crude drug/human/day.

Experimental example 2: rat Chronic Atrophic Gastritis (CAG) model experiment

2.1 Experimental methods

2.1.1 animal groups

50 SPF-grade healthy male Wistar rats are selected as animals, and the laboratory temperature is controlled to be 26 +/-2 ℃ and the relative humidity is maintained to be 40-60% in the whole experiment process. After 1 week of acclimatization, 10 animals were randomly selected as blank control group and the remaining 40 animals were subjected to CAG molding. After the molding was successful, 40 samples were randomly grouped into high, medium and low dose groups and model groups of 10 samples each.

2.1.2 CAG rat model preparation

Wistar rats were given a solution of MNNG at a concentration of 120. mu.g/ml and 0.1% ammonia water daily for free drinking, the above solution was replaced every 24 hours, and the process of solution preparation was performed in a dark environment. Feeding granulated rat feed containing 0.03% ranitidine; feeding for hunger and satiety disorder (2 days of hunger and 1 day of hunger), and feeding to stomach with 40% ethanol solution when fasting.

2.1.3 methods of administration

The high, medium and low dose fractions of the sample were administered by gavage with 20 g/kg (equivalent to 20 g/kg of crude drug, the same below), 10 g/kg (equivalent to 10 g/kg of crude drug, the same below), and 5 g/kg (equivalent to 5 g/kg of crude drug, the same below), respectively. The normal control group and the model group were administered with equal volumes of purified water, respectively. The gavage is continued for 15 days.

2.1.4 method for obtaining materials and preparation of specimen

After the administration of the drug, the experimental mice take fasting measures, but are maintained for 24 hours by normal drinking water. Injecting the chloral hydrate solution into the abdominal cavity of the rat for anesthesia, fixing the laboratory rat after the drug effect is exerted, opening the abdominal cavity, and taking out the complete stomach. The stomach is longitudinally split along the greater curvature, and after the stomach is washed by normal saline to remove residues in the stomach, the tissue of the lesser curvature of the stomach is cut by a sharp blade and is placed into paraformaldehyde solution for tissue fixation.

Paraffin sections were prepared and HE stained by conventional methods. The pathological changes of the stomach tissues of the animals in each group are observed by a microscope.

2.2 pathological section HE staining results

The HE staining results of all groups of pathological sections show that the stomach tissue mucosa lamina propria in the blank animal stomach tissue visual field is rich in stomach glands, closely arranged, normal in structure and normal in stomach gland epithelial cell morphology (see figure 5). The natural gland arrangement of the stomach tissue of the model group animal is loose, the natural layer of the gastric mucosa is seriously congested, and a large number of inflammatory cells are arranged under the mucosa with edema (see figure 6). Compared with the model group, the pathological changes of the stomach tissues of the animals in the high dose group (20 g crude drug/kg) and the medium dose group (10 g crude drug/kg) of the sample group are obviously improved. The concrete expression is as follows: the inherent glands are arranged more closely, and the number of the glands is not obviously reduced; submucosal inflammatory cells were significantly reduced with slight edema (see fig. 7 and 8).

2.3 conclusion

The above results show that: the traditional Chinese medicine composition prepared by the invention has the effect of improving the chronic atrophic gastritis of rats. The lowest effective dose is 10 g crude drug/kg, and the dose is converted into human clinical equivalent dose of 100 g crude drug/human/day.

Experimental example 3: clinical experiments

The traditional Chinese medicine formula of the invention is mainly used for treating various stomach diseases caused by the insufficiency of gastric acid and digestive enzyme, such as functional dyspepsia, gastrointestinal hypomotility, chronic superficial gastritis accompanied with dyspepsia or bile reflux, chronic atrophic gastritis, hypertrophic gastritis, or stomach diseases accompanied with erosion, polyp, variola, wart, macula, macular tumor, etc., and cardia inflammation, reflux esophagitis, hiatal hernia, Barrett esophagus, polyp, etc. caused by the stomach diseases.

3.1 medical record selection: clinically, 1.5 ten thousand patients with gastropathy diagnosed by a gastroscope and treated according to the formula of the invention, 8923 male patients and 6077 female patients, wherein the age range is 13-85 years and the average age is 54 years. The case distribution is as follows: 1372 cases of functional dyspepsia, 9754 cases of chronic superficial gastritis accompanied by dyspepsia, 2310 cases of chronic atrophic gastritis, 68 cases of hypertrophic gastritis, 6129 cases accompanied by bile reflux, 12980 cases accompanied by gastrointestinal hypomotility, 1457 cases accompanied by erosion, 982 cases of polyps, 186 cases of pockmarks, 245 cases of warts, 132 cases of macula or macular tumors, and 361 cases of cardia inflammation, 629 cases of reflux esophagitis, 164 cases of hiatal hernia, 108 cases of Barrett esophagus, 234 cases of polyps, etc. caused by such stomach diseases.

3.2 methods of treatment: and the above 16 types of diseases, the patients are randomly divided into 16 groups, the medicinal granule preparation prepared in the embodiment 1 of the invention (can be replaced by other preparation forms), the dosage of the traditional Chinese medicine is one day, each 15 days is a treatment course, and other diseases are treated for 4 treatment courses except for 6 treatment courses of atrophic gastritis.

3.3 treatment outcome: after a course of treatment, 81% of patients have obvious clinical relief of various symptoms; after three to four treatments, the clinical symptom relieving rate is 96 percent. Patients with atrophic gastritis need to continue recovery treatment of pathological changes for one month after the clinical symptoms have been reduced or disappeared. After the treatment course is finished, the result is checked again by the gastroscope, and the gastroscope and pathological results of 76% of patients are obviously improved or recovered to be normal. Some of the typical cases are as follows:

(1) grandpa, 68 years old, for nearly 10 years, begins to experience epigastric pain in the morning after meals, with pain mild and severe. When the patient does not eat the meal at night, the pain of the gastric cavity can not occur; if the user eats dinner, the stomach pain will affect the sleep. In order to relieve the epigastric pain, people have to eat less or no meal, so that the body becomes thin day by day, diseases such as anemia and myocardial ischemia occur, and uncomfortable feelings such as palpitation, hypodynamia, dizziness and the like often occur. The chronic atrophic gastritis with variola and macular tumor is obtained as a result of gastroscopy. Many western medicines and traditional Chinese medicines are taken intermittently, so that the disease condition is not improved all the time and gradually worsens. After the formula disclosed by the invention is taken for 6 treatment courses, the pain in the gastric cavity disappears, and the weight is increased by 9 jin; the results of the repeated examination with gastroscope show that the stomach atrophy, the variola and the macular tumor disappear and return to normal. The diseases of anemia and myocardial ischemia and the uncomfortable feelings of palpitation, hypodynamia, dizziness and the like are also reviewed to be recovered to be normal.

(2) Mr. Huang, man, 34 years old, has been known to suffer from epigastric fullness during treatment for nearly 2 years. Always feel that the stomach is blocked flusterly and the stomach is not ventilated up and down, sometimes feel hard when pressing the stomach and is buckled on a bowl, and feel oppressed and uncomfortable on the abdomen when the stomach is serious; he does not eat the rice when he says that the stomach and the intestine are not comfortable all day long, and very trouble is felt. The result of 3 gastroscopies was chronic gastritis and bile reflux. The traditional Chinese medicine and western medicine are taken all the time, and no obvious effect is achieved. After the formula is taken for treating one course of treatment, the epigastric fullness and oppression are obviously relieved; after the treatment is continued for three courses of treatment, the epigastric fullness is not attacked, and the bile reflux disappears after the gastroscope is carried out to review.

(3) In women, the age of 39 years old, gastric cavity is noisy for more than 7 years, the feeling of messy grass in the stomach fossa is often seen, the discomfort feeling can occur even if people eat inappropriate things, and the feeling of discomfort can not be realized. Performing gastroscopy for many times, and the result is that the chronic atrophic gastritis is accompanied with bile reflux, erosion and polyp; the patients have taken more medicines and are not good all the time. After the formula is taken for 6 treatment courses, gastric cavity noise disappears; the results of the repeated examination with a gastroscope show that the stomach atrophy, bile reflux, erosion and polyp disappear and return to normal.

(4) Mr. forest, man, age 58, who says he has hiccup for more than 5 years at the time of pulse diagnosis, and the disease is seen everywhere in the country, and the hiccup is not good all the time after taking medicine all the year round. The result of multiple physical examinations and gastroscopy is chronic atrophic gastritis. The hiccup sound is low and frequent at ordinary times, and basically hiccups except the sleeping time in one day; he feels that he has qi in the throat and frequently goes out. After the formula is taken for 2 treatment courses, hiccup is improved obviously; after conditioning for 6 treatment courses, hiccup bothering Mr. Cheng has disappeared, and gastroscopy shows that atrophic gastritis has recovered to normal.

(5) The diagnosis is that no appetite is present for more than 1 year at ordinary times and chronic atrophic gastritis and erosion are caused by gastroscopy. The medicine is taken for treatment all the time, and no obvious improvement is realized. After the traditional Chinese medicine is taken for treatment, the effect is obvious after 4 treatment courses, and the feeling of unwilling to eat is not achieved; the treatment is continued for 2 courses of treatment, and the gastric atrophy and erosion disappear after the gastroscope is performed.

(6) Houda Daye, 73 years old, has got the disease of heartburn for more than 20 years when feeling pulse, and how many times the stomach is examined, the chronic gastritis begins, then becomes atrophic gastritis with bile reflux, erosion and wart, and the medicine for treating stomach disease is taken all the time, has no effect on heartburn and is increasingly serious. In recent years, heartburn and back pain occur frequently in the middle of the night, and the fever, dysphoria and sleep incapability of the whole body caused by burning are caused seriously, so that the treatment has no effect and is very painful. After the formula disclosed by the invention is taken for 1 treatment course, the feeling of heartburn and fever is obviously improved; after conditioning for 6 treatment courses, heartburn puzzling Hou recovers, and gastroscopy shows that atrophic gastritis, bile reflux, erosion and wart also disappear and recover to be normal.

3.4 the above results show that: the traditional Chinese medicine composition prepared by the invention has obvious curative effect on treating gastropathy caused by insufficient secretion of gastric acid and digestive enzyme.

Experimental example 4: liquid phase analysis

4.1 sample treatment: the unfermented and fermented liquid medicine of example 1 was taken, 1 mL of the liquid medicine was sucked up by a pipette gun into a 2 mL centrifuge tube, centrifuged at 13000 r/min for 3 min, and the supernatant was taken and filtered through a 0.45 μm pinhole filter as a sample.

4.2 chromatographic conditions:

(1) a chromatographic column: eclipse XDB-C18 (4.6 mm. times.250 mm, 5 μm);

(2) mobile phase species: acetonitrile (B) -water (A) is used as a mobile phase;

(3) detection wavelength: 285 nm;

(4) column temperature: 30 ℃;

(5) flow rate: 0.8 mL min-1;

(6) sample introduction volume: 10 mu L of the solution;

(7) gradient elution ratio: see table 3.

TABLE 3

Time (min)	Flow Rate (mL. min)^-1）	Acetonitrile (B%)	Ultrapure water (A%)
				0	0.8	5	95
15	0.8	15	85
				35	0.8	60	40
50	0.8	5	95
				51	0.8	5	95

4.3 conclusion: comparing fig. 1 and fig. 2, it can be seen that the concentration of some components of the traditional Chinese medicine composition after fermentation is significantly reduced, for example, the retention time is at a characteristic peak around 26 min, and the concentration after fermentation is significantly reduced, so that the method can effectively realize the micromolecule formation of the traditional Chinese medicine through microbial fermentation. From the peak output quantity, the quantity of certain characteristic peaks of the fermented traditional Chinese medicine composition is obviously reduced, and the toxic components of the traditional Chinese medicine can be decomposed or structurally modified by proving the decomposition and transformation capacity of microorganisms, so that the toxic and side effects of the traditional Chinese medicine are reduced.

Experimental example 5: capillary electrophoresis analysis

5.1 sample treatment: 60 mL each of the unfermented and post-fermented solutions of example 1 were placed in two 250 mL beakers, which were placed on a magnetic stirrer at room temperature. Then slowly adding saturated ammonium sulfate solution to make the concentration of saturated ammonium sulfate reach 20%, stirring for 60 min. And after the centrifugation is finished, centrifuging for 30 min at 4000 r/min, and separating the supernatant from the precipitate after the centrifugation. The pellet was washed with 0.02mol/LPBS buffer (pH = 7.4) and retained, and the resulting supernatant was subjected to the above procedure. Adding ammonium sulfate to reach 70% saturation degree, mixing the obtained protein cleaning solution, placing in dialysis bag with relative molecular mass of 3500 KDa for 24h, and desalting. After the desalting is finished, the obtained protein mixed solution of the unfermented liquid medicine and the fermented liquid medicine is centrifuged for 30 min at 4000 r/min, and the supernatant is taken and filtered by a 0.45 mu m pinhole filter membrane to be used as a sample.

5.2 chromatographic conditions:

(1) a chromatographic column: TOSOH TSKgelG3000SWXL (7.8 mm. times.300 mm, 5 μm);

(2) mobile phase: 25mmol/LPBS buffer (pH = 6.8);

(3) detection wavelength: 280 nm;

(4) column temperature: 25 ℃;

(5) flow rate: 1 mL min-1;

(6) sample introduction volume: 20 μ L (sample concentrated to 1 mg/mL);

(7) elution procedure: isocratic elution;

(8) recording time: and (5) 25 min.

5.3 conclusion: as can be seen from a comparison of FIG. 3 and FIG. 4, the amount of the macromolecular protein (molecular weight > 670 KDa) present in the unfermented liquor is 45.61%; the proportion of macromolecular protein (molecular weight is more than 670 KDa) in the fermented liquid medicine is obviously reduced and is only 5.99 percent.

Claims

1. A traditional Chinese medicine composition for treating chronic benign gastropathy and esophageal diseases is characterized by being prepared from the following raw medicinal materials in parts by weight: 8-12 parts of codonopsis pilosula, 8-12 parts of fried bighead atractylodes rhizome, 8-12 parts of poria cocos, 4-8 parts of fingered citron, 4-8 parts of immature bitter orange, 4-8 parts of hovenia dulcis thumb, 4-8 parts of sea buckthorn, 4-8 parts of perilla leaf, 4-8 parts of dark plum, 4-8 parts of seville orange flower, 4-8 parts of ginger processed pinellia and 8-12 parts of liquorice.

2. The traditional Chinese medicine composition for treating chronic benign gastropathy and esophageal disease according to claim 1, which is characterized by being prepared from the following raw medicinal materials in parts by weight: 9-11 parts of codonopsis pilosula, 9-11 parts of fried bighead atractylodes rhizome, 9-11 parts of poria cocos, 5-7 parts of fingered citron, 5-7 parts of immature bitter orange, 5-7 parts of hovenia dulcis thumb, 5-7 parts of sea buckthorn, 5-7 parts of perilla leaf, 5-7 parts of dark plum, 5-7 parts of seville orange flower, 5-7 parts of ginger processed pinellia tuber and 9-11 parts of liquorice.

3. The traditional Chinese medicine composition for treating chronic benign gastropathy and esophageal disease according to claim 2, which is characterized by being prepared from the following raw medicinal materials in parts by weight: 10 parts of codonopsis pilosula, 10 parts of fried bighead atractylodes rhizome, 10 parts of poria cocos, 6 parts of fingered citron, 6 parts of immature bitter orange, 6 parts of hovenia dulcis thunb, 6 parts of sea buckthorn, 6 parts of perilla leaf, 6 parts of dark plum, 6 parts of seville orange flower, 6 parts of pinellia ternate and 10 parts of liquorice.