CN113413402A - Application of plum blossom extract in preparation of medicine for treating helicobacter pylori infection diseases - Google Patents
Application of plum blossom extract in preparation of medicine for treating helicobacter pylori infection diseases Download PDFInfo
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- CN113413402A CN113413402A CN202110880525.9A CN202110880525A CN113413402A CN 113413402 A CN113413402 A CN 113413402A CN 202110880525 A CN202110880525 A CN 202110880525A CN 113413402 A CN113413402 A CN 113413402A
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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)
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- 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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- 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 or decoction
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- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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Abstract
The invention discloses an application of a plum blossom extract in preparing a medicament for treating helicobacter pylori infection diseases, belongs to the technical field of medicaments, determines that the plum blossom extract can inhibit the toxicity and the planting capability of Hp in vitro for the first time, obtains the plum blossom extract through animal experiment verification that can effectively inhibit the infection and the inflammation state of Hp, has a single medicament application effect superior to or equal to the treatment effect of triple or quadruple, and can synergistically improve the eradication effect after Hp infection and the gastric mucosa protection effect by combining the plum blossom extract with the triple or quadruple; the plum blossom extract is adopted to treat Hp infection diseases, so that the cost is low, the treatment course is short, and the treatment is convenient; the invention provides a new thought for treating Hp infectious diseases and has great significance for clinical treatment and popularization of Hp.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to an application of a plum blossom extract in preparation of a medicine for treating helicobacter pylori infection diseases.
Background
Flos mume (Parnasis pallustris L.) is the whole herb or root of the plant of the genus Mesembryana of the family Saxifragaceae, i.e., the plant Plectranthus niveus (L.) Nees, also called Plectranthus niveus, California scabra, etc., and is a special Mongolian medicine commonly used by Mongolian generations and folks, and Mongolian names are Cannabis, Wuli Di Ge, and Nichiyan Nichigan-its grid, etc. Is one of the common Mongolian medicinal materials used for treating tumors and related diseases in modern Mongolian medicine clinical application. The plum blossom is sweet and bitter in flavor and free of obvious toxicity, has the effects of clearing heat and removing toxicity, breaking distension and fullness, relieving swelling and expelling pus, inhibiting festering and the like, is mainly clinically used for treating visceral heat festering and heat fullness diseases, namely treating liver and gall tumors, partial intestinal tumors (such as colon tumors) and the like, and can also be used for treating icteric hepatitis, sphagitis, parotitis, angiitis, tuberculosis and the like.
Helicobacter pylori (Hp) is a microaerophilic gram-negative bacterium that colonises the human stomach extensively. More than 50% of people in the world are infected by the helicobacter pylori infection, and researches prove that the helicobacter pylori infection is closely related to the high incidence rate of chronic gastritis, gastric ulcer, duodenal ulcer and gastric cancer. Currently, the main clinical methods for treating helicobacter pylori include triple or quadruple therapy, i.e., 2 or 3 antibiotics in combination with 1 proton pump inhibitor or bismuth agent. The special stomach environment and the emptying of the stomach lead to large dosage, long treatment course, high cost and large side effect of helicobacter pylori treatment, so that the compliance of patients is poor, partial patients stop taking the medicine after not reaching the complete treatment course, in addition, the helicobacter pylori is easy to generate drug resistance to the used antibiotics, even 6 antibiotic resistant strains appear, and the helicobacter pylori is easy to re-infect. Therefore, the radical treatment of helicobacter pylori infection has become a medical problem. The traditional Chinese medicine compound preparation has good curative effect on helicobacter pylori, but has unclear components, long treatment course, inconvenient administration and higher cost than western medicines.
At present, no report related to the preparation of the medicament for treating the helicobacter pylori infection diseases by using the plum blossom extract is found.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides the application of the plum blossom extract in preparing the medicines for treating the helicobacter pylori infection diseases.
In order to achieve the purpose, the invention provides the following technical scheme:
the invention provides an application of a plum blossom extract in preparing a medicament for treating helicobacter pylori infection diseases.
Further, the helicobacter pylori infection disease is gastritis, gastric ulcer, duodenal ulcer or gastric cancer.
Further, the preparation method of the plum blossom grass extract comprises the following steps:
preparation of the organic phase extract of the plum blossom grass: heating and reflux-extracting flos mume with ethanol to obtain extractive solution, removing ethanol from the extractive solution, and drying to obtain organic phase extract;
preparation of aqueous extract of plum blossom grass: heating and leaching flos mume with water, filtering to obtain leaching solution, concentrating, and drying the obtained concentrated solution to obtain flos mume water phase extract;
mixing the plum blossom organic phase extract and the plum blossom aqueous phase extract in equal mass to obtain the plum blossom extract.
Further, in the preparation process of the organic phase extract of the plum blossom, the ethanol is 95 vol.% ethanol, during heating reflux extraction, the material-liquid ratio of the plum blossom to the ethanol is 1g (1-5 mL), the heating temperature is 90-95 ℃, the extraction is carried out for 4h each time, the extraction is carried out for 3 times, the obtained extracting solutions are combined for 3 times, and the drying is spray drying.
Further, the spray drying is carried out in a spray drying tower, the air inlet temperature of the spray drying tower is 120-150 ℃, the air outlet temperature of the spray drying tower is 90-95 ℃, the atomization frequency is 220-300 Hz, the feeding frequency is 4-6.5 Hz, and the induced air frequency is 20-50 Hz.
Spray drying the ethanol extract of flos mume can retain the inherent aromatic substances and heat sensitive active ingredients of flos mume.
Further, in the preparation process of the aqueous extract of the plum blossom, when the plum blossom is heated and extracted by using water, the material-liquid ratio of the plum blossom to the water is 1g to (5-10 mL), the heating temperature is 90-95 ℃, the extraction time is 1-2h, the concentration is specifically reduced pressure concentration at 50-60 ℃, the drying temperature is 65-75 ℃, and the heating and extraction of the plum blossom is preferably performed by using distilled water.
Furthermore, the plum blossom extract is used as an effective active ingredient and added with pharmaceutically acceptable auxiliary materials to be prepared into tablets, capsules, granules, oral liquid, suspending agents or injection, and the mass of the plum blossom extract in various formulations accounts for 30-60% of the total mass of the medicine.
Compared with the prior art, the invention has the following beneficial effects:
the toxicity and the planting capacity of the plum blossom extract for inhibiting Hp in vitro are determined for the first time, animal experiments prove that the plum blossom extract can effectively inhibit the infection and the inflammation aggravation of Hp to mouse gastric mucosa, the effect of single application is superior to or equivalent to the triple or quadruple treatment effect, and the triple or quadruple treatment effect can be synergistically improved after the Hp is infected;
the plum blossom extract is adopted to treat Hp infection diseases, so that the cost is low, the treatment course is short, and the treatment is convenient;
the invention provides a new idea for treating Hp infection diseases and has great significance for clinical treatment of Hp.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 is a statistical chart of relative expression amounts of Hp-related virulence and colonization genes (gag A, Vac A, Bab A) in the Hp group and the Medinieria speciosa treatment group;
FIGS. 2(a) and 2(b) are schematic external views of gastric mucosa of mice infected with Hp and treated with plum blossom grass;
FIG. 3 is a graph of HE staining of gastric mucosa in various groups of mice;
FIG. 4 is a transmission electron micrograph of the mitochondrial structure of the gastric mucosa of each group of mice;
FIG. 5 is a transmission electron micrograph of the microvilli of the gastric mucosa of each group of mice.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention. It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.
Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The description and examples are intended to be illustrative only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
The following examples used Medinioides that were collected from Wula national nature protection area, Sikhan, Chifeng city, inner Mongolia autonomous region, and the whole drug was dried in the shade, and the drug was identified as Medinioides (Parnasasalusterillinn) by professor assisting in the study of Xuan Zhan, university student of pharmacy, inner Mongolian, and professor Irina, institute of Mongolian medicine.
The helicobacter pylori standard strain SS1 adopted by the invention is preserved and presented by a laboratory of the digestive system department of the first hospital of Beijing university; the male Kunming mouse (KM mouse) is 6 weeks old and has a body weight of 36-41g, and is purchased from Beijing Wittingli laboratory animal technology Limited company, and the laboratory animal production license is as follows: beijing Baishan SCXK (Beijing) 2016-; license for use of experimental animal: beijing SYXK (Beijing) 2017-; the triple drug employed: rabeprazole sodium enteric-coated tablets: medical industry (specification 10mg x 7); clarithromycin tablets: shanghai yapei (specification 250mg × 8); amoxicillin capsules: the Zhuhai Federal (specification 500mg 48/24), the four-drug combination adopted, on the basis of the above three-drug combination, is added with colloidal bismuth pectin capsules: shanxi mante (specification 100mg 48);
the description will not be repeated below.
Example 1
The preparation of the plum blossom extract comprises the following steps:
(1) organic phase extraction of the plum blossom grass: taking 0.40kg of dried whole flos Lupuli, heating to 90 deg.C with 400ml of 95 vol.% ethanol, reflux-extracting for 3 times, each for 4 hr, mixing the extractive solutions obtained by 3 times, and volatilizing ethanol to obtain flos Lupuli ethanol extract; drying the extract in a spray drying tower to obtain the organic phase extract of the plum blossom grass, wherein the air inlet temperature of the spray drying tower is 120 ℃, the air outlet temperature is 90 ℃, the atomization frequency is 220Hz, the feeding frequency is 4Hz, and the induced air frequency is 20Hz, so that the inherent aromatic substances and heat-sensitive active ingredients of the plum blossom grass can be well reserved.
(2) Water-phase extraction of the plum blossom grass: weighing 25g of dried plum blossom grass, pulverizing by a pulverizer, injecting 200ml of deionized water, leaching in a water bath at 90 ℃ for 1 hour, filtering, concentrating at 60 ℃ under reduced pressure to obtain a concentrated solution, sterilizing the obtained water-phase extract by using a 0.22um filter, and drying the filtered concentrated solution in a 75 ℃ oven to obtain the plum blossom grass water-phase extract for later use;
(3) and (3) mixing the plum blossom organic phase extract obtained in the step (1) and the plum blossom aqueous phase extract obtained in the step (2) in equal mass to obtain the plum blossom extract.
Example 2
Preparation of the plum blossom extract:
(1) organic phase extraction of the plum blossom grass: taking 0.40kg of dried flos Lupuli whole plant, heating to 95 deg.C with 2000ml of 95 vol.% ethanol, reflux-extracting for 3 times, each for 4 hr, mixing the extractive solutions obtained by 3 times, and volatilizing ethanol to obtain flos Lupuli ethanol extract; drying the extract in a spray drying tower to obtain the organic phase extract of the plum blossom grass, wherein the air inlet temperature of the spray drying tower is 150 ℃, the air outlet temperature is 95 ℃, the atomization frequency is 300Hz, the feeding frequency is 5Hz, and the induced air frequency is 30Hz, so that the inherent aromatic substances and heat-sensitive active ingredients of the plum blossom grass can be well reserved.
(2) Water-phase extraction of the plum blossom grass: weighing 25g of dried plum blossom grass, pulverizing by a pulverizer, injecting 125ml of deionized water, leaching in a water bath at 95 ℃ for 1 hour, filtering, concentrating at 50 ℃ under reduced pressure to obtain a concentrated solution, sterilizing the obtained water-phase extract by using a 0.22um filter, and drying the filtered concentrated solution in a drying oven at 65 ℃ to obtain the plum blossom grass water-phase extract for later use;
(3) and (3) mixing the plum blossom organic phase extract obtained in the step (1) and the plum blossom aqueous phase extract obtained in the step (2) in equal mass to obtain the plum blossom extract.
Example 3
Preparation of the plum blossom extract:
(1) organic phase extraction of the plum blossom grass: taking 0.40kg of dried whole flos Lupuli, heating to 90 deg.C with 1000ml of 95 vol.% ethanol, reflux-extracting for 3 times, each for 4 hr, mixing the extractive solutions obtained by 3 times, and volatilizing ethanol to obtain flos Lupuli ethanol extract; drying the extract in a spray drying tower to obtain flos mume organic phase extract, wherein the air inlet temperature of the spray drying tower is 130 deg.C, the air outlet temperature is 90 deg.C, the atomization frequency is 250Hz, the feeding frequency is 6.5Hz, and the induced air frequency is 50Hz, so as to retain inherent aromatic substances and thermosensitive active ingredients of flos mume.
(2) Water-phase extraction of the plum blossom grass: weighing 25g of dried plum blossom grass, pulverizing by a pulverizer, injecting 250ml of deionized water, leaching in a 90 ℃ water bath for 2 hours, filtering, concentrating at 55 ℃ under reduced pressure to obtain a concentrated solution, sterilizing the obtained water phase extract by using a 0.22um filter, and drying the filtered concentrated solution in a 70 ℃ oven to obtain the plum blossom grass water phase extract for later use;
(3) and (3) mixing the plum blossom organic phase extract obtained in the step (1) and the plum blossom aqueous phase extract obtained in the step (2) in equal mass to obtain the plum blossom extract.
Effect test example 1
Agar dilution method for determining Minimum Inhibitory Concentration (MIC) of flos mume extract
The MIC of the Mesona extract was determined by agar dilution as follows: scraping cultured Hp for 48-72 hr, inoculating in liquid culture medium, and diluting to 1.5 x 10 with sterile physiological saline8CFU/mL. 200uL of the bacterial solution was dropped onto the surface of the medium, and the mixture was uniformly inoculated onto drug-containing plates of different final concentrations of Mesona extracts (40g/L, 20g/L, 10g/L, 5g/L, 2.5g/L, 1.25g/L) using a coating rod, and then subjected to microaerophilic culture at 37 ℃ for 72 hours, where MIC was the lowest drug concentration at which Hp growth did not occur. MIC determination results were judged according to the American Committee for clinical laboratory standardization (NCCLS).
When MIC was measured by the agar dilution method, Hp was not observed in the 20g/L, 10g/L plum blossom-containing plate, i.e., 10g/L was the MIC value. The results suggest that the plum blossom extract has a certain anti-Hp activity.
Effect test example 2
Real-time quantitative PCR detection of influence of plum blossom grass extract on expression of Hp colonization adhesion related gene
Hp infects human body and adheres to gastric mucosa cells by expressing outer membrane proteins such as Bab A, and induces inflammation and cell injury by expressing proteins such as Vac A and Cag A, so as to increase the permeability of gastric mucosa and facilitate the migration of gastric mucosa. Bab A is Hp outer membrane protein gene, can bind host cell receptor specifically, give Hp permanent planting ability; the Hp strain with high toxicity has a cytotoxin related gene (gag A) and is involved in the transport of the gag A and the inflammatory reaction of a host. Vacuolar cytotoxin A (Vac A) can destroy endocytosis, and inhibit immune cells to cause immune tolerance and chronic infection, so the primer design of the related genes is carried out, the primer design is shown in table 1, and the influence of the plum blossom extract on the Hp virulence and colonization is evaluated.
Dissolving the Mesona extract with distilled water to a final concentration of 20 mg/ml. Resuspend the cultured Hp broth to 1 x 10 with PBS7CFU/ml, 0.1ml of the PBS suspension of Hp was added to the prepared plum blossom extract solution, incubated at 37 ℃ with shaking microaerophilic for 24 hours, centrifuged to discard the supernatant, washed 3 times with PBS, and total RNA was isolated using RNA prepure Cell/Bacteria Kit (Tiangen, China). cDNA Synthesis kit (SYBR @ primeScript)TMRT-PCR Kit) operation. The primer sequences are shown in Table 1. The PCR reaction system is as follows: 2ul of cDNA, 25ul of UltraSYBR mix, 1ul of 10uM upstream and downstream primers, respectively, were made up to 50ul with deionized water. Reaction conditions of 95 ℃, 10min, 95 ℃, 10s, 60 ℃, 1min and 40 cycles, adopting 2-△△CTThe relative expression level of the target gene is calculated by the method. The relative expression level of the target gene in the two groups was compared by using Hp bacterial suspension without adding the plum blossom extract solution as a control group (the treatment method was the same as above).
TABLE 1
Using 16S rRNA gene as internal reference, 2-△△CTThe differences in expression of the genes involved in the establishment of the Hp colonization after the action of the control group and the Medinieria extract were analyzed, and the results are shown in FIG. 1. As can be seen from FIG. 1, the extract of the plum blossom plant inhibits the above-mentioned Bab A, Cag A and Vac A virulence gene expression, therefore, it is speculated that the Mesona chinensis can reduce the colonization and damage of Hp to gastric mucosa by inhibiting the expression of Hp-related colonization and virulence genes.
Effect test example 3
Animal modeling and group dosing observation
Preparation of a suspension of helicobacter pylori: a microaerophilic culture method is adopted, gas reaches microaerophilic conditions (85% nitrogen, 10% carbon dioxide and 5% oxygen) in microaerophilic environment, helicobacter pylori strains are inoculated on a solid plate containing 7% horse serum, and per 100ml of solid Hp culture medium (20 g of the Hp solid culture medium: Merck campylobacter jejuni culture medium, 500ml of distilled water is added for dissolving, 6.81kg of autoclaving is carried out for 15min, 50ml of sheep blood and antibiotic selection solution are introduced after cooling to about 50 ℃, and after uniform mixing, a sterile culture plate is poured, 1ml of microbial additives are added, and after 48 hours of constant temperature culture at 37 ℃, the culture is carried out. After being identified as positive by fast urease, oxidase, smear gram stain and catalase, bacteria are collected by an inoculating loop and evenly mixed in a bacteria transfer base (the bacteria transfer base is prepared by adding 6.75g of bovine heart brain extract into 500ml of distilled water for dissolving, sterilizing under high pressure for 15min at 6.81kg, cooling to 4 ℃ and subpackaging for later use), and the concentration of a turbidimetric tube is controlled to be 1 x 109CFU/ml for use.
The experimental method comprises the following steps: 132 mice of KM mice were randomly divided into 8 groups, each of which was a healthy group (hereinafter, also referred to as "normal group") 12 mice completely untreated, a normal Saline group (salt group) 12 mice, an Hp group 18 mice, a triple group 18 mice, a quadruple group 18 mice, a Medinioway group 18 mice, a triple + Medinioway group 18 mice, a quadruple + Medinioway group 18 mice, a first day, the Hp group, the triple group, the quadruple group, the Medinioway group, the triple + Medinioway group and the quadruple + Medinioway group mice were treated with helicobacter pylori suspension in an amount of 400. mu.L/mouse, and then, the mice were treated with drugs once every other day, each time was 200. mu.L/mouse, after 4 weeks, 3 mice in each group were selected, the infection state of Hp was evaluated, and after confirming that each group was successfully modeled, the mice in each group were started to be administered with drugs (healthy group completely untreated, physiological Saline group was not administered, and only normal Saline group was administered with normal Saline) In the later stage of Hp group, only physiological saline, only triple drug, only quadruple drug, only plum blossom extract, and the triple and plum blossom groups, and the quadruple and plum blossom extracts, and the quadruple and plum blossom groups, respectively, were administered simultaneously. Wherein, the dosage of the plum blossom extract is as follows: the dosage was calculated according to standard adult Mesona extract 2000mg bid, based on the body weight of Kunming mouse. The dosage of the triple drug is as follows: specifically, 70kg of adult triple-usage dosage (20 mg of rabeprazole, 1000mg of amoxicillin and 500mg of clarithromycin) is converted by referring to clinical standard adult dosage. Administration amount of the tetrad drug: quadruple therapy on an adult basis (rabeprazole 20mg bid, amoxicillin 1000mg bid, clarithromycin 500mg bid + colloidal bismuth pectin 300mg bid) was performed in accordance with body weight conversion of standard adult (70kg) and KM mice.
Each group of mice are administrated at intervals of one day, 1 time is administrated every day, the medicines are placed in water and stirred every time of administration to obtain 200 mu L of liquid medicine, the liquid medicine is administrated by gastric administration after fasting and water prohibition are carried out for 12 hours, the water is supplied and fasted after the latter hour of administration, the continuous administration is carried out for 5 times, then KM mice are sacrificed, serum and anatomical stomach tissues are left for relevant experimental detection, a Rapid Urease Test (RUT) and the immunohistochemical staining of HE and Hp are carried out, and ELISA detection and transmission electron microscope analysis on the oxidative stress indexes in the serum are carried out.
The gastric mucosa of the mice in the Hp group and the plum blossom grass group are observed, as shown in fig. 2(a) and fig. 2(b), respectively, as can be seen from fig. 2, the gastric mucosa of the mice in the Hp group has obvious bleeding and congestion, while the gastric mucosa of the plum blossom grass extract group has smooth surface, and has no erosion, bleeding and ulcer.
The gastric mucosa of each group of mice was HE stained and visualized (100X), as shown in fig. 3: the normal group has clear gastric mucosa structure, and the Hp infected group can show mucosa abscission, erosion and thinning. The triple and quadruple groups have a certain protection effect, the plum blossom group can obviously increase the thickness of the gastric mucosa, protect and promote the repair of the gastric mucosa, and the plum blossom extract combines the triple and quadruple groups to more synergistically protect the structure of the gastric mucosa.
The gastric mucosa helicobacter pylori infection of each group of mice was examined by the rapid urease method (RUT), and the results are shown in Table 2.
TABLE 2
As can be seen from Table 2, the plum blossom extract has good treatment effect on helicobacter pylori infection, the quality effect of the single plum blossom extract on the helicobacter pylori infection of the mouse stomach is superior to the treatment effect of the triple drug, and the cure rate can be effectively improved by combining the plum blossom extract with the triple drug or the quadruple drug.
The mitochondria of gastric mucosal cells and the microvilli of gastric mucosa of each group of mice were observed by transmission electron microscopy, and the results are shown in fig. 4 and 5, respectively. FIG. 4 shows: the normal group can see that the mitochondrial structure is complete, the internal ridge is clear, the mitochondrial structure can be broken after Hp infection, the internal ridge is disordered, the triple and quadruple groups can repair the mitochondrial structure to a certain extent, the plum blossom extract also has a protection effect on the mitochondrial structure, the plum blossom extract can effectively protect the mitochondrial function when combined with the triple and quadruple groups, and the result of figure 4 indicates that the plum blossom extract can singly or jointly protect the mitochondrial structure of the gastric mucosal epithelial cells after Hp infection. FIG. 5 shows: the normal group of visible microvilli are regularly arranged and have a longer structure. After Hp infection, microvilli were found to break and disappear, and were recovered to different degrees by triple or quadruple therapy. The single plum blossom extract can also better recover the microvilli structure, and when the plum blossom extract is combined with the triple or quadruple, the microvilli structure is better as a whole. The results in fig. 5 suggest that the plum blossom extract alone or in combination with triplets or quadruplets can protect the microvilli structure of the gastric mucosal epithelial cells after Hp infection.
The serum IL-2, IL-8 and TNF-a of each group of mice were assayed by enzyme-linked immunosorbent assay (ELISA) and the results are shown in Table 3 in units of: pg/mg.
TABLE 3
Note:*：p＜0.05，**：p＜0.01。
as can be seen from table 3: through serum ELISA detection, the expression levels of IL-2, IL-8 and TNF-a are obviously increased in an Hp group, while the levels of related inflammatory factors are obviously reduced in a Mesembryanthemum gracile extract treatment group, compared with a pure Hp infection group, the levels of related inflammatory factors are statistically different (p is less than 0.05), and the anti-inflammatory and anti-oxidation effects are more obvious in a Mesembryanthemum gracile extract combined triple and quadruple treatment group.
The results of the above-described effect tests on the plum blossom extract prepared in example 2 and example 3 were performed by the same methods as in effect test example 1, effect test example 2, and effect test example 3, and were consistent with the effects of the plum blossom extract of example 1.
The above description is only for the preferred embodiment of the present invention, and the protection scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical scope of the present invention, the technical solution and the inventive concept of the present invention equivalent or change within the technical scope of the present invention.
Claims (6)
1. Application of flos mume extract in preparing medicine for treating helicobacter pylori infection is provided.
2. The use according to claim 1, wherein the helicobacter pylori infection disease is gastritis, gastric ulcer, duodenal ulcer or gastric cancer.
3. The use as claimed in claim 1, wherein the preparation method of the plum blossom extract comprises the steps of:
preparation of the organic phase extract of the plum blossom grass: heating and reflux-extracting flos mume with ethanol to obtain extractive solution, removing ethanol from the extractive solution, and drying to obtain organic phase extract of flos mume;
preparation of aqueous extract of plum blossom grass: heating and leaching flos mume with water, filtering to obtain leaching solution, concentrating, and drying the obtained concentrated solution to obtain flos mume water phase extract;
mixing the plum blossom organic phase extract and the plum blossom aqueous phase extract in equal mass to obtain the plum blossom extract.
4. The use as claimed in claim 3, wherein the preparation of the organic phase extract of flos mume comprises 95 vol.% ethanol, the heating reflux extraction comprises extracting flos mume and ethanol at a ratio of 1g to (1-5 mL) at 90-95 deg.C for 4h for 3 times, mixing the extractive solutions obtained by 3 times, and spray drying.
5. The use of claim 3, wherein in the preparation of the aqueous extract of plum blossom, when the plum blossom is extracted with water under heating, the ratio of plum blossom to water is 1g (5-10 mL), the heating temperature is 90-95 ℃, the extraction time is 1-2h, the concentration is specifically under reduced pressure at 50-60 ℃, and the drying temperature is 65-75 ℃.
6. The use as claimed in claim 1, wherein the said extract of plum blossom is used as active ingredient to be added with pharmaceutically acceptable adjuvants to make into tablet, capsule, granule, oral liquid, suspension or injection, and the mass of plum blossom in various dosage forms is 30-60% of the total mass of the medicament.
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