CN110151938B - New application of vine tea preparation - Google Patents
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- CN110151938B CN110151938B CN201910565421.1A CN201910565421A CN110151938B CN 110151938 B CN110151938 B CN 110151938B CN 201910565421 A CN201910565421 A CN 201910565421A CN 110151938 B CN110151938 B CN 110151938B
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/076—Poria
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/87—Vitaceae or Ampelidaceae (Vine or Grape family), e.g. wine grapes, muscadine or peppervine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/906—Zingiberaceae (Ginger family)
- A61K36/9062—Alpinia, e.g. red ginger or galangal
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
Abstract
The invention belongs to the field of traditional Chinese medicine health products, and particularly relates to a new application of a vine tea preparation. The ampelopsis grossedentata preparation is a compound preparation consisting of four raw materials of ampelopsis grossedentata, galangal, poria cocos and mung beans, has a certain protection effect on rats with alcoholic gastric mucosa injury models, can obviously reduce the gastric juice amount of rats with alcoholic gastric mucosa injury, increases the pH value of gastric juice, relieves ulcer injury, and has a better effect of protecting alcoholic gastric mucosa injury. In addition, the ampelopsis grossedentata preparation has the advantages of low cost, simple preparation process, convenience in carrying and taking and the like.
Description
Technical Field
The invention belongs to the field of traditional Chinese medicine health products, and particularly relates to a new application of a vine tea preparation.
Background
Excessive drinking can cause acetaldehyde in human body not to be completely decomposed, so that drunk symptoms such as nausea, vomiting, coma and discomfort are generated after people drink the wine. Manifested by physical symptoms such as fatigue, headache, thirst, vertigo, gastropathy, nausea, emesis, insomnia, hand tremor and increased or decreased blood pressure; mental symptoms include acute anxiety, irritability, oversensitivity, depression, or feelings of guilt. Chronic gastritis is caused by long-term drinking. The incidence of gastric mucosa injury disease of drinking people is obviously higher than that of non-drinking people. Gastric mucosal damage is associated with factors such as gastric acid hypersecretion, infection with helicobacter pylori (h. pylori), inhibition of Cyclooxygenase (COX) activity by non-steroidal anti-inflammatory drugs (NSAIDs), and oxidative stress caused by Reactive Oxygen Species (ROS) produced. Alcohol is one of the key factors of gastric mucosa injury, and the main mechanisms of the injury of the gastric mucosa by the alcohol comprise: alcohol can interfere gastric mucosa secretion activity, change cell permeability and consume gastric mucus, and the pH value of normal gastric juice can be reduced; alcohol can induce lipid peroxidation of gastric mucosa, generate oxygen free radicals, and damage gastric mucosa.
The invention discloses a liver-protecting and stomach-regulating composition, a preparation method and application thereof (CN 201910124081.9). The liver-protecting and stomach-regulating composition is a health-care product consisting of coix seeds, sweet chestnuts, radish seeds, platycodon grandiflorum, dwarf lilyturf roots, schisandra chinensis, bitter almonds and liquorice. However, the health product composition disclosed in the invention has complex components, and the price of part of the medicines is higher, so that the prepared health product has higher cost and higher price, and cannot be popularized to the public. Doctor thesis "protection of astragalus-pueraria drug against ethanol-induced gastric mucosal injury" (2015), the protective effect of astragalus-pueraria drug against ethanol-induced gastric mucosal injury was studied, wherein astragalus-pueraria 3: the compatibility of the medicines 1 has better effect on protecting gastric mucosa injury than other compatibility groups, and can reduce pathological injuries such as gastric mucosa hemorrhage, edema, erosion and the like caused by ethanol.
The inventor carries out a great deal of research in the early period, and applies for an anti-intoxication, anti-alcoholism and liver protection preparation and a preparation process thereof (CN201410751437.9) in 12 months and 10 days in 2014, and the invention takes medicinal and edible health food or food as raw materials to prepare the anti-intoxication, anti-alcoholism and liver protection preparation which has good functions of anti-intoxication, anti-alcoholism and liver protection. The preparation for preventing drunkenness, dispelling the effects of alcohol and protecting liver is a compound preparation consisting of four raw materials of vine tea, galangal, tuckahoe and mung beans. The ampelopsis grossedentata is a natural product, has small toxic and side effects, contains a large amount of flavonoids, and has special effects in relieving ethanol poisoning, preventing alcoholic liver and fatty liver, inhibiting hepatocyte deterioration, reducing incidence of liver cancer and protecting liver; galangal is pungent and hot in nature, enters spleen and stomach channels, has the effects of dispelling cold, relieving pain, warming middle energizer and arresting vomiting, is recorded in the new herbal medicine, has the effects of warming stomach, dispelling cold, helping digestion, dispelling alcohol effect and treating stomach protein cold pain and has a certain effect of dispelling the effects of alcohol; poria has effects of promoting diuresis, eliminating dampness, invigorating spleen, calming heart, promoting discharge of alcohol in human body with urine, and relieving hangover. The mung bean has the efficacy of clearing heat and removing toxicity, and can effectively relieve the symptoms of dizziness, vomiting of characters and the like after drinking.
The inventor unexpectedly finds that the ampelopsis grossedentata preparation has a certain protection effect on an alcoholic gastric mucosa injury model rat, can obviously reduce the gastric juice amount of the alcoholic gastric mucosa injury model rat, increases the pH value of the gastric juice, relieves ulcer injury, and has a better effect of protecting the alcoholic gastric mucosa injury. Therefore, the invention provides the application of the vine tea preparation in preparing the medicine or the health-care product for preventing or treating the alcoholic gastric mucosa injury.
Disclosure of Invention
The invention aims to provide a new application of a vine tea preparation.
The technical scheme adopted by the invention is as follows:
the application of a vine tea preparation in preparing a medicine or a health-care product for preventing or treating gastric mucosal injury is characterized in that the vine tea preparation is prepared from the following raw materials in parts by weight: 20-50 parts of vine tea, 5-15 parts of galangal, 10-40 parts of mung bean and 10-40 parts of poria cocos.
Preferably, the ampelopsis grossedentata preparation consists of the following raw materials in parts by weight: 30-50 parts of vine tea, 5-15 parts of galangal, 15-35 parts of mung bean and 10-40 parts of poria cocos.
Preferably, the ampelopsis grossedentata preparation consists of the following raw materials in parts by weight: 30 parts of vine tea, 5 parts of galangal, 30 parts of mung bean and 35 parts of poria cocos.
Preferably, the ampelopsis grossedentata preparation consists of the following raw materials in parts by weight: 35 parts of vine tea, 10 parts of galangal, 30 parts of mung bean and 25 parts of poria cocos.
Preferably, the ampelopsis grossedentata preparation consists of the following raw materials in parts by weight: 30 parts of vine tea, 15 parts of galangal, 35 parts of mung bean and 20 parts of poria cocos.
Preferably, the ampelopsis grossedentata preparation consists of the following raw materials in parts by weight: 40 parts of vine tea, 5 parts of galangal, 25 parts of mung bean and 30 parts of poria cocos.
Preferably, the gastric mucosal lesion is alcoholic gastric mucosal lesion.
The application of the ampelopsis grossedentata preparation in preparing the drugs or the health care products for preventing or treating the alcoholic gastric mucosa injury, wherein the preparation method of the ampelopsis grossedentata preparation comprises the following steps:
a. weighing the raw materials according to a formula;
b. adding Ampelopsis grossedentata into water with the mass 5-15 times of that of Ampelopsis grossedentata, soaking for 10-60min, reflux-extracting at 50-100 deg.C for 1-3 times, each time for 30-120min, filtering with 200 mesh filter cloth after each reflux-extraction, mixing filtrates, and refrigerating at 4 deg.C for 24 hr to obtain material A;
c. centrifuging the material A in the step B for 5-15min under the condition of 2000-4000r/min to obtain a precipitate and a supernatant, washing the precipitate with water with the mass of 0.1-2 times that of the supernatant, centrifuging for 5-15min under the condition of 2000-4000r/min, repeating for 3-5 times, and finally drying the obtained precipitate at 60-100 ℃ to obtain a material B;
d. the preparation method comprises the steps of uniformly mixing poria cocos, galangal and mung beans to obtain a mixed material, adding the mixed material into water with the mass 5-15 times of that of the mixed material, performing reflux extraction at 50-100 ℃ for 1-3 times, performing reflux extraction for 30-120min each time, filtering through 200-mesh filter cloth after each reflux extraction, combining filtrates, concentrating, and drying at 60-100 ℃ to obtain a material C.
e. And (D) uniformly mixing the material B obtained in the step (C) with the material C obtained in the step (D) to obtain a mixed material D, and preparing a corresponding preparation by using the mixed material D as an active ingredient and adopting pharmaceutically conventional auxiliary materials and processes.
Preferably, the ampelopsis grossedentata preparation is any one of chewable tablets, capsules and granules.
Preferably, the ampelopsis grossedentata preparation is a granule.
The invention has the beneficial effects that: compared with a positive control group 1 (Lizhudele), a positive control group 2 (cimetidine) and a positive control group 3 (astragalus-kudzu root traditional Chinese medicine liquid) and a positive control group 4 (galangal), the ampelopsis grossedentata preparation claimed by the invention can obviously reduce the gastric juice amount of a rat with alcoholic gastric mucosa injury, increase the pH value of gastric juice, relieve ulcer injury and have a better effect of protecting the alcoholic gastric mucosa injury; meanwhile, the ampelopsis grossedentata preparation has the advantages of convenience in carrying and taking, low cost, simple production process and the like.
Detailed Description
The present invention will be specifically explained below with reference to specific embodiments, but the scope of the present invention is not limited to the following examples.
Example 1A Ampelopsis grossedentata preparation for preventing or treating gastric mucosal injury 1
A vine tea preparation for preventing or treating gastric mucosal injury is prepared from the following raw materials in parts by weight: 30 parts of vine tea, 5 parts of galangal, 30 parts of mung bean and 35 parts of poria cocos.
The preparation process of the ampelopsis grossedentata preparation comprises the following steps:
a. weighing the raw materials according to a formula;
b. soaking Ampelopsis grossedentata in water 5 times the mass of Ampelopsis grossedentata for 60min, reflux-extracting at 50 deg.C for 1 time for 30min, filtering with 200 mesh filter cloth, and cold-preserving the filtrate at 4 deg.C for 24 hr to obtain material A;
c. centrifuging the material A in the step B for 15min under the condition of 2000r/min to obtain a precipitate and a supernatant, washing the precipitate with water with the mass of 0.1 time that of the supernatant, centrifuging for 15min under the condition of 4000r/min, repeating for 3 times, and finally drying the obtained precipitate at 80 ℃ to obtain a material B;
d. uniformly mixing Poria cocos, rhizoma Alpiniae Officinarum and mung bean to obtain a mixed material, adding the mixed material into water 15 times of the mass of the mixed material, performing reflux extraction at 100 ℃ for 3 times, performing reflux extraction for 30min each time, filtering through 200-mesh filter cloth after each reflux extraction, combining filtrates, concentrating, and drying at 100 ℃ to obtain a material C;
e. uniformly mixing the material B obtained in the step C with the material C obtained in the step D to obtain a mixed material D;
d. the chewable tablet is prepared by the following formula and process by weight percent: preparing 50% of the mixed material D, 40% of soluble starch and 9% of cane sugar into soft materials by using ethanol, granulating, drying, granulating, adding 1% of magnesium stearate, and pressing into chewable tablets for preventing or treating gastric mucosal injury.
Example 2 Ampelopsis grossedentata preparation for preventing or treating gastric mucosal injury 2
A vine tea preparation for preventing or treating gastric mucosal injury is prepared from the following raw materials in parts by weight: 35 parts of vine tea, 10 parts of galangal, 30 parts of mung bean and 25 parts of poria cocos.
The preparation process of the ampelopsis grossedentata preparation comprises the following steps:
a. weighing the raw materials according to a formula;
b. adding ampelopsis grossedentata into water with the mass 10 times of that of the ampelopsis grossedentata, soaking for 10min, performing reflux extraction at 80 ℃ for 3 times, performing reflux extraction for 120min each time, filtering through 200-mesh filter cloth after reflux extraction for each time, combining filtrates, and refrigerating at 4 ℃ for 24h to obtain a material A;
c. centrifuging the material A in the step B for 10min under the condition of 4000r/min to obtain a precipitate and a supernatant, washing the precipitate with water with the mass 1 time that of the supernatant, centrifuging for 10min under the condition of 2000r/min, repeating for 5 times, and finally drying the obtained precipitate at 60 ℃ to obtain a material B;
d. the preparation method comprises the steps of uniformly mixing the poria cocos, the galangal and the mung beans to obtain a mixed material, adding the mixed material into water with the mass being 10 times of that of the poria cocos, the galangal and the mung beans, performing reflux extraction for 1 time at 80 ℃, performing reflux extraction for 60min, filtering through 200-mesh filter cloth after the reflux extraction, concentrating a filtrate, and drying at 80 ℃ to obtain a material C.
e. Uniformly mixing the material B obtained in the step C with the material C obtained in the step D to obtain a mixed material D;
d. the tablet is prepared according to the following formula and process by weight percent: preparing the mixture D with 70% and starch with 29% into soft mass with ethanol, granulating, drying, grading, adding 1% of pulvis Talci, and pressing into tablet for preventing or treating gastric mucosa injury.
Example 3A Ampelopsis grossedentata preparation for preventing or treating gastric mucosal injury 3
A vine tea preparation for preventing or treating gastric mucosal injury is prepared from the following raw materials in parts by weight: 30 parts of vine tea, 15 parts of galangal, 35 parts of mung bean and 20 parts of poria cocos.
The preparation process of the ampelopsis grossedentata preparation comprises the following steps:
a. weighing the raw materials according to a formula;
b. soaking Ampelopsis grossedentata in water 15 times the mass of Ampelopsis grossedentata for 40min, reflux-extracting at 100 deg.C for 3 times, each time for 80min, filtering with 200 mesh filter cloth after each reflux extraction, mixing filtrates, and refrigerating at 4 deg.C for 24h to obtain material A;
c. centrifuging the material A in the step B for 5min under the condition of 3000r/min to obtain a precipitate and a supernatant, washing the precipitate with water 2 times the mass of the supernatant, centrifuging for 5min under the condition of 4000r/min, repeating for 4 times, and finally drying the obtained precipitate at 100 ℃ to obtain a material B;
d. the preparation method comprises the steps of uniformly mixing the poria cocos, the galangal and the mung beans to obtain a mixed material, adding the mixed material into water with the mass 5 times of that of the poria cocos, the galangal and the mung beans, performing reflux extraction for 3 times at 50 ℃, performing reflux extraction for 120min each time, filtering through 200-mesh filter cloth after each reflux extraction, combining filtrates, concentrating, and drying at 60 ℃ to obtain a material C.
e. Uniformly mixing the material B obtained in the step C with the material C obtained in the step D to obtain a mixed material D;
d. the granules are prepared according to the following formula and process by weight percent: preparing the 30% mixed material D, 55% dextrin and 15% sucrose into soft materials by using ethanol, granulating, drying and finishing granules to obtain the granules for preventing or treating gastric mucosal injury.
Example 4A Ampelopsis grossedentata preparation for preventing or treating gastric mucosal injury 4
A vine tea preparation for preventing or treating gastric mucosal injury is prepared from the following raw materials in parts by weight: 40 parts of vine tea, 5 parts of galangal, 25 parts of mung bean and 30 parts of poria cocos.
The preparation process of the ampelopsis grossedentata preparation comprises the following steps:
a. weighing the raw materials according to a formula;
b. soaking Ampelopsis grossedentata in water 15 times the mass of Ampelopsis grossedentata for 40min, reflux-extracting at 100 deg.C for 2 times, each time for 80min, filtering with 200 mesh filter cloth after each reflux extraction, mixing filtrates, and refrigerating at 4 deg.C for 24h to obtain material A;
c. centrifuging the material A in the step B for 5min under the condition of 3000r/min to obtain a precipitate and a supernatant, washing the precipitate with water 2 times the mass of the supernatant, centrifuging for 5min under the condition of 3000r/min, repeating for 4 times, and finally drying the obtained precipitate at 80 ℃ to obtain a material B;
d. the preparation method comprises the steps of uniformly mixing the poria cocos, the galangal and the mung beans to obtain a mixed material, adding the mixed material into water with the mass 5 times of that of the poria cocos, the galangal and the mung beans, performing reflux extraction for 2 times at 80 ℃, performing reflux extraction for 120min each time, filtering through 200-mesh filter cloth after each reflux extraction, combining filtrates, concentrating, and drying at 100 ℃ to obtain a material C.
e. Uniformly mixing the material B obtained in the step C with the material C obtained in the step D to obtain a mixed material D;
d. and preparing the mixed material D into a soft material by using ethanol, granulating, drying, and filling into hard capsules to obtain the capsules for preventing or treating gastric mucosal injury.
Example 5 study of Ampelopsis grossedentata preparation for preventing alcoholic gastric mucosal injury in mice
1. Drug configuration
Ampelopsis grossedentata preparation 1: the formula ratio of the preparation in example 1, and granules prepared according to the method in example 3; ampelopsis grossedentata preparation 2: the formula ratio of the preparation in example 2, and granules prepared according to the method in example 3; ampelopsis grossedentata preparation 3: granules prepared according to the method of example 3, in the formula ratio described in example 3; ampelopsis grossedentata preparation 4: granules prepared according to the method of example 3 and in the formula ratio of example 4; lizhudele: preparing the Lizhudele into a suspension; astragalus-kudzuvine root traditional Chinese medicine liquid: mixing radix astragali and radix Puerariae at a weight ratio of 3:1, decocting with 10 times of water for 3 times, each time for 30min, and mixing decoctions to obtain radix astragali and radix Puerariae Chinese medicinal liquid; galangal group: weighing rhizoma Alpiniae Officinarum with the same amount as the raw materials in each Ampelopsis Grossdentata preparation, adding into 5 times of water, reflux-extracting at 50 deg.C for 3 times, each for 120min, filtering with 200 mesh filter cloth, mixing filtrates, concentrating, and drying at 60 deg.C to obtain rhizoma Alpiniae Officinarum extract, and formulating with normal saline when it is used.
2. Grouping and administration of drugs
180-220g, 100 male Wistar rats (purchased from animal testing center, lanzhou biologicals ltd) were randomly divided into 10 groups, control group: 10ml/kg of physiological saline; model group: after the normal saline 10ml/kg is perfused for 14 days, perfusing 1 ml/one of 56-degree red star Erguotou; vine tea group: 1 vine tea preparation group (the formula is described in example 1, the dosage is 1g/kg based on crude drug amount), 2 vine tea preparation groups (the formula is described in example 2, the dosage is 1g/kg based on crude drug amount), 3 vine tea preparation groups (the formula is described in example 3, the dosage is 1g/kg based on crude drug amount), 4 vine tea preparation groups (the formula is described in example 4, the dosage is 1g/kg based on crude drug amount), and after the stomach is filled for 14 days, the stomach is filled by 56 degrees of Hongxing Erguotou, and the dosage is the same as that of the model group; positive control group 1: gavage suspension, 1g/kg of gavage suspension, and then performing gavage on a 56-degree red star Erguotou for 14 days, wherein the dosage is the same as that of a model group; positive control group 2: after the cimetidine is infused in 10ml/kg for 14 days, the cimetidine is infused in 56-degree red star Erguotou, and the dosage is the same as that of the model group; positive control group 3: the administration amount of the astragalus-kudzuvine root traditional Chinese medicine liquid is 1g/kg based on crude drug amount, after the stomach is perfused for 14 days, the stomach is perfused by 56 degrees of red star Erguotou, and the dosage is the same as that of a model group; galangal group: the galangal rhizome extract has the administration amount of 1g/kg based on crude drug amount, and after the gavage for 14 days, the gavage is carried out by 56-degree red star Erguotou, and the dose is the same as that of the model group.
3. Treatment of
Before the last administration, rats are fasted for 24h, water is forbidden for 8h, 1h after the last administration, 0.35ml/kg of each group of rats is subjected to intraperitoneal injection of 10% chloral hydrate for anesthesia, blood is taken from abdominal aorta, serum is obtained by centrifugation, the whole stomach is taken, gastric juice is collected and the pH value of the gastric juice is measured, a general specimen is observed, and the rats are placed at the temperature of minus 20 ℃ for standby.
4. Index measurement
4.1 body weight determination
Body weights of rats were measured daily before administration.
4.2 measurement of gastric juice and gastric acid value in rats
And pouring out the gastric contents in a graduated centrifuge tube, collecting gastric juice, and observing the degree and the form of the injury of the gastric mucosa. Centrifuging the collected gastric juice in a centrifuge at 1500r/min for 10min, collecting supernatant, and determining total acidity by titration method.
4.3 gastric tissue ulcer index detection
After the stomach was removed, the stomach was cut open along the greater curvature, the size of the ulcer was measured with a ruler and magnifying glass and the ulcer index was calculated according to the Guth standard: marking 1 point for speckle erosion; erosion length < lmm 2 points; 3 minutes are counted from 1mm to 2 mm; 4 minutes are counted from 2mm to 3 mm; the value is multiplied by 2 when the mark is more than 3mm, 5 minutes and the width is more than 1 mm.
4.4 semi-quantitative analysis of gross Pathology
According to the proportion of the lesion area in the whole gastric mucosa, the diagnosis is semiquantitative, namely no abnormality (score 0), slight lesion (score 1), slight lesion (score 2), moderate lesion (score 3) and severe lesion (score 4).
4.5 histopathological Observation and Scoring
After the gross examination, the parts with the most severe gastric mucosal lesions of each animal were cut out, fixed in 10% formaldehyde solution, and subjected to routine flaking, HE staining and microscopic observation. The normal cross section of the gastric mucosa, including the observation of the full layer of the mucosa, was selected. According to the fact that the depth of the mucosa layer is affected by the lesion as a main basis for judging the severity of the mucosa lesion, the lesion is classified into no abnormality (score 0), slight lesion (score 1: mucosa damaged depth < mucosa layer 1/3), slight lesion (score 2: mucosa layer 1/3 < mucosa damaged depth < mucosa layer 2/3), moderate lesion (score 3: mucosa damaged depth > mucosa layer 2/3) and severe lesion (score 4: lesions such as mucosa lesion not only affects the whole layer, but also visible hemorrhage of the mucosa layer) to observe the type of the lesion.
4.6 measurement of levels of NO, SOD, MDA and GSH in serum
And (3) taking the serum obtained by centrifugation in the step (3), and measuring the levels of NO (nitric oxide), SOD (superoxide dismutase), MDA (malondialdehyde) and GSH (glutathione) in the rat serum by adopting a serum kit.
4.7 measurement of mRNA expression of eNOS (endothelial nitric oxide synthase), nNOS (neuronal nitric oxide synthase) and COX-2 (cyclooxygenase 2) in stomach tissue
Collecting the above prepared stomach tissue, cutting, homogenizing with physiological saline to obtain 10% stomach tissue homogenate, centrifuging to obtain supernatant, extracting total RNA in stomach tissue with Trizol, measuring total RNA concentration with micro spectrophotometer, and diluting RNA stock solution to 1 μ g/mL. Taking 1 mu L of diluted RNA solution, adding 1 mu L of (oligo) Primer dT and 10 mu L of sterilized water for injection, reacting for 5min at 65 ℃, taking out, adding 4 mu L of Reaction Buffer, 1 mu L of Riblock RNase Inhibitor and 2 mu L of dNTP mix, mixing uniformly, adding 1 mu L of reverse Aid M-mu/v RT, and synthesizing cDNA at 42 ℃, 60min, 70 ℃ and 5 min. Then, 10. mu.L Master, 1. mu.L forward primer, 1. mu.L reverse primer, 1. mu.L cDNA template and 7. mu.L sterile water for injection were added to the octaplex tube, and centrifuged to mix well. And finally, putting the mixture into a qPCR instrument for amplification, wherein the amplification conditions are as follows: denaturation at 95 deg.C for 15min, annealing at 60 deg.C for 1h, extension at 95 deg.C for 15min, and then circulating 40 times. Using housekeeping gene GAPDH as reference gene, passing through 2-ΔΔCTThe relative expression level of each gene was calculated.
5. Statistical analysis
All data are expressed as mean ± standard deviation, and the significance analysis of differences was performed using SPSS 17.0 software, with P <0.05 indicating significant differences.
6. Results
6.1 rat body weight changes
The weight of rats in each group before gastric lavage is not different (P is more than 0.05), after 14 days of administration, the weight of rats in each group is obviously increased (P is less than 0.05) compared with that before administration, but no obvious difference (P is more than 0.05) exists among groups, which indicates that the ampelopsis grossedentata preparation has no toxic or side effect, does not influence the diet of rats and does not influence the normal growth of rats as the other groups, and the results are shown in Table 1.
TABLE 1 weight change of rats in each group before and after administration
Note:*p is less than 0.05, and is compared with the weight of the rat before gastric lavage
6.1 measurement of gastric juice and gastric acid values in rats
Gastric mucosal damage is exacerbated by either hypersecretion of gastric fluid or by too low a pH of the gastric fluid. As can be seen from Table 2, the gastric juice amount of rats in the model group is the largest, and the gastric juice amounts of rats with the gavage ampelopsis grossedentata preparation and the positive control drug are both significantly lower than those of rats in the model group (P is less than 0.05), wherein the gastric juice amount of rats in the ampelopsis grossedentata preparation 3 group is closer to that of the control group; the pH value of gastric juice of rats in the model group is the lowest, the pH values of gastric juice of rats filled with the ampelopsis grossedentata preparation and the positive control drug are both obviously lower than that of rats in the model group (P is less than 0.05), and the pH value of gastric juice of rats in the ampelopsis grossedentata preparation 3 group is closer to that of rats in the control group. The ampelopsis grossedentata preparation can reduce the excessive secretion of the stomach fluid of the rat and can maintain the normal pH value in the stomach of the rat, so that the damage of alcohol to the gastric mucosa can be reduced to a certain extent.
TABLE 2 gastric juice and gastric acid values in rats
Note:*p is less than 0.05, compared with a control group;#p is less than 0.05, compared with the model group
6.2 gastric tissue ulcer index detection, gross observation score and histopathology score
The gastric tissue ulcer index detection shows that: the control group rats had smooth gastric mucosa surface without ulcer and ulcer index of 0; the stomach tissues of the rats in the model group can be seen with a plurality of linear erosions with the length of more than 3mm, the width of more than 1mm and the ulcer index of (8.8 +/-1.40); the ampelopsis grossedentata preparation group and the positive control group are improved compared with the model group, wherein the ulcer indexes of the ampelopsis grossedentata preparation 2, the ampelopsis grossedentata preparation 3 and the positive control group rats are obviously lower than that of the model group (P is less than 0.05), and the ulcer index of the ampelopsis grossedentata preparation 3 group rats is lower than that of the positive control group.
Gross observation score: the control group of rats has smooth gastric mucosa surface, no congestion and erosion focus and no abnormality; the gastric mucosa of the model group rat can be seen with a plurality of gastric mucosa punctate, linear erosion and bleeding, and the general observation score is (3.5 +/-0.71); the ampelopsis grossedentata preparation group and the positive control group are improved compared with the model group, wherein the gross observation score of the ampelopsis grossedentata preparation 2, the ampelopsis grossedentata preparation 3 and the positive control group is obviously lower than that of the model group (P <0.05), and the gross observation score of the ampelopsis grossedentata preparation 3 group is lower than that of the positive control group.
And (3) histopathological scoring: the lesions are classified into non-abnormal lesions (score 0), slight lesions (score 1: mucosa damaged depth < mucosa layer 1/3), slight lesions (score 2: mucosa layer 1/3 < mucosa damaged depth < mucosa layer 2/3), moderate lesions (score 3: mucosa damaged depth > mucosa layer 2/3) and severe lesions (score 4: lesions such as mucosa damaged not only involve the whole layer, but also visible bleeding of the mucosa), and the types of the lesions are observed. The control group of rats has smooth gastric mucosa surface, no congestion and erosion focus and no abnormality; most of the damaged depths of the gastric mucosa of the rats in the model group are greater than the mucosal layer 2/3, and the histopathological score is (3.6 +/-0.52); the ampelopsis grossedentata preparation group and the positive control group are improved compared with the model group, wherein the histopathology scores of the ampelopsis grossedentata preparation 2, the ampelopsis grossedentata preparation 3 and the positive control group rats are obviously lower than that of the model group (P is less than 0.05), and the histopathology scores of the ampelopsis grossedentata preparation 3 group rats are lower than that of the positive control group.
The index detection of gastric tissue ulcer, gross observation score and histopathology score show that the ampelopsis grossedentata preparation can play a certain role in inhibiting the damage of the gastric mucosa of a rat caused by alcohol, wherein the scoring result is shown in table 3.
TABLE 3 gastric tissue ulcer index, gross observation score and histopathological score for each group of rats
Note:#p is less than 0.05, compared with the model group
6.3 measurement of levels of NO, SOD, MDA, and GSH in serum
From the results in table 4, the NO content, SOD activity and GSH content in the serum of the model rat are all significantly lower than those of the control group (P <0.05), and the MDA content is significantly higher than that of the control group (P < 0.05); the NO content, SOD activity and GSH content in the blood serum of rats in the ampelopsis grossedentata preparation and a positive control group are all obviously higher than those in a model group (P <0.05), and the MDA content is obviously lower than that in the model group (P <0.05), wherein the NO content, SOD activity, GSH content and MDA content of rats in the ampelopsis grossedentata preparation 3 group are closer to the level of rats in the control group compared with those in the positive control group. The ampelopsis grossedentata preparation can increase the NO content, SOD activity and GSH content in rat serum, reduce the MDA content and further protect stomach tissues from oxidative damage caused by alcohol.
TABLE 4 serum NO, SOD, MDA, GSH levels in rats of each group
Note:*p is less than 0.05, compared with a control group;#p is less than 0.05, compared with the model group
6.4 measurement of mRNA expression of eNOS, nNOS and COX-2 in stomach tissue
Gastric ulcer development is associated with excessive inhibition of eNOS and nNOS, and alcoholic gastric mucosal damage leads to increased expression of COX-2. The relative expression of the inflammatory factors related to the rat gastric tissue in each group in the experiment is shown in table 5, the expression water of eNOS and nNOS in the rat gastric tissue of the model group is obviously lower than that of the control group (P <0.05), and the expression level of COX-2 is obviously higher than that of the control group (P < 0.05); the expression water of eNOS and nNOS in the ampelopsis grossedly higher than that of a model group (P <0.05) and the expression water of COX-2 is obviously lower than that of the model group (P <0.05) in the stomach tissues of rats in the ampelopsis grossedentata preparation and a positive control group; wherein, compared with the positive control group, the expression levels of eNOS, nNOS and COX-2 in the stomach tissues of the rats in the ampelopsis grossedentata preparation 3 group are closer to the level of the rats in the control group. The result shows that the ampelopsis grossedentata preparation can up-regulate the expression of eNOS and nNOS in the stomach tissue of a rat with alcoholic gastric mucosa injury and down-regulate the expression of COX-2, and has a certain protective effect on the gastric mucosa injury caused by alcohol.
TABLE 5 expression levels of eNOS, nNOS, and COX-2 in the gastric tissues of various groups of rats
Note:*p is less than 0.05, compared with a control group;#p is less than 0.05, compared with the model group
7. Conclusion
According to the invention, the protection effect of the ampelopsis grossedentata preparation disclosed by the invention on the gastric mucosa injury is discussed by taking the alcoholic gastric mucosa injury of a rat as a model, and the result shows that the ampelopsis grossedentata preparation disclosed by the invention can obviously reduce the gastric juice amount of the alcoholic gastric mucosa injury rat, increase the pH value of the gastric juice, relieve the ulcer injury and have a better effect of protecting the alcoholic gastric mucosa injury. The alcoholic gastric mucosa injury is the most common gastric mucosa injury model, and on the basis that the ampelopsis grossedentata preparation disclosed by the invention has a prevention or treatment effect on the gastric mucosa injury caused by alcohol, the ampelopsis grossedentata preparation also has a certain prevention or treatment effect on the gastric mucosa injury caused by other factors, and the method can be understood by those skilled in the art.
Claims (5)
1. The application of the ampelopsis grossedentata preparation in preparing the medicine or the health-care product for improving gastric mucosal injury is characterized in that the ampelopsis grossedentata preparation is prepared from the following raw materials in parts by weight: 30 parts of vine tea, 15 parts of galangal, 35 parts of mung bean and 20 parts of poria cocos.
2. The use of ampelopsis grossedentata preparation as claimed in claim 1 for the preparation of a medicament or health product for ameliorating gastric mucosal damage, wherein said gastric mucosal damage is alcoholic gastric mucosal damage.
3. The use of the ampelopsis grossedentata preparation as claimed in claim 1 in the preparation of a medicament or health product for improving gastric mucosal lesion, wherein the ampelopsis grossedentata preparation method comprises the following steps:
a. weighing the raw materials according to a formula;
b. adding Ampelopsis grossedentata into water with the mass 5-15 times of that of Ampelopsis grossedentata, soaking for 10-60min, reflux-extracting at 50-100 deg.C for 1-3 times, each time for 30-120min, filtering with 200 mesh filter cloth after each reflux-extraction, mixing filtrates, and refrigerating at 4 deg.C for 24 hr to obtain material A;
c. centrifuging the material A in the step B for 5-15min under the condition of 2000-4000r/min to obtain a precipitate and a supernatant, washing the precipitate with water with the mass of 0.1-2 times that of the supernatant, centrifuging for 5-15min under the condition of 2000-4000r/min, repeating for 3-5 times, and finally drying the obtained precipitate at 60-100 ℃ to obtain a material B;
d. uniformly mixing Poria cocos, rhizoma Alpiniae Officinarum and mung bean to obtain a mixed material, adding the mixed material into water with the mass 5-15 times of that of the mixed material, performing reflux extraction at 50-100 ℃ for 1-3 times, performing reflux extraction for 30-120min each time, filtering through 200-mesh filter cloth after each reflux extraction, combining filtrates, concentrating, and drying at 60-100 ℃ to obtain a material C;
e. and (D) uniformly mixing the material B obtained in the step (C) with the material C obtained in the step (D) to obtain a mixed material D, and preparing a corresponding preparation by using the mixed material D as an active ingredient and adopting pharmaceutically conventional auxiliary materials and processes.
4. The use of a Ampelopsis grossedentata preparation as claimed in claim 3 in the preparation of a medicament or health product for ameliorating gastric mucosal lesion, wherein the Ampelopsis grossedentata preparation is any one of a tablet, a capsule and a granule.
5. The use of the Ampelopsis grossedentata preparation as claimed in claim 4 in the preparation of a medicament or health product for improving gastric mucosal lesion, wherein the Ampelopsis grossedentata preparation is in the form of granules.
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