CN111297979B

CN111297979B - Application of Shibi tea and/or camellin A in preparation of medicine for treating gastric ulcer

Info

Publication number: CN111297979B
Application number: CN202010283110.9A
Authority: CN
Inventors: 袁尔东; 练颖仪; 张文姬; 孙世利; 黎秋华; 孙伶俐; 赖幸菲; 陈若虹
Original assignee: South China University of Technology SCUT; Tea Research Institute Guangdong Academy of Agricultural Sciences
Current assignee: South China University of Technology SCUT; Tea Research Institute Guangdong Academy of Agricultural Sciences
Priority date: 2020-04-10
Filing date: 2020-04-10
Publication date: 2021-06-08
Anticipated expiration: 2040-04-10
Also published as: CN111297979A

Abstract

The invention belongs to the technical field of application of Shibi tea (Adinandranitida) and an active monomer thereof, and particularly relates to application of Shibi tea and an active monomer Camellianin A (Camellia A, CA) thereof in preparation of a medicine for treating gastric ulcer. The key protection of the invention is the application of the Shibi tea and the camellin A in the preparation of the medicine for treating gastric ulcer; and the application of the Shibi tea or the camellin A in preparing the medicine for treating the gastric ulcer, wherein the camellin A is extracted from the Shibi tea. The animal acute gastric ulcer model experiment proves that the aqueous extract of the Shiqian tea and the active monomer camellin A thereof have good treatment effect on gastric ulcer, and the expression of related inflammatory factors IL-6 is down-regulated by regulating the NF kappa B inflammation regulation signal path, so that the injury degree and inflammatory reaction of gastric ulcer are reduced.

Description

Application of Shibi tea and/or camellin A in preparation of medicine for treating gastric ulcer

Technical Field

The invention belongs to the technical field of application of Shibi tea and an active monomer thereof, and particularly relates to application of Shibi tea and an active monomer camellianin A thereof in preparation of a medicine for treating gastric ulcer.

Background

Gastric ulcer refers to damage of the gastric mucosa caused by acidic digestive injury, which often occurs in the gastric horn, antrum, cardia, etc., and is one of peptic ulcers, and also a common and frequent gastrointestinal disease. Patients with gastric ulcer usually have epigastric pain, such as burning sensation, and typical dyspepsia symptoms, such as abdominal distension, nausea, fullness, heartburn and the like, and even have serious symptoms of peptic ulcer complications, such as bleeding, perforation or gastric outlet obstruction and the like, so that the quality of life and the quality of work of the patients are greatly influenced.

At present, there are many kinds of therapeutic drugs for gastric ulcer in clinic, mainly including histamine receptor antagonists such as cimetidine and ranitidine, and proton pump inhibitors such as omeprazole, etc. These drugs are not only expensive, but also cause side effects such as diarrhea, constipation, fatigue, drowsiness, headache, and muscle pain, and even acute liver injury. Therefore, the method has important practical significance for exploring efficient and safe natural active substances from plant resources to improve and treat the gastric ulcer.

The existing researches on gastric ulcer by plant resources or natural active substances are mostly related to prevention, the treatment effect of the gastric ulcer is less researched, and the gastric ulcer is mostly a formula and a compound traditional Chinese medicine with health care effect. The formula has various components, complex proportion and tedious processing, and certain traditional Chinese medicine components such as ginseng, lucid ganoderma and the like have expensive price. Therefore, it is important to find common natural plants or highly effective and safe natural active ingredients with the effect of treating gastric ulcer.

The invention technology that single plant extracts or monomers have good drug treatment effect on gastric ulcer is less concerned at present. Generally, researches on prevention and treatment of gastric ulcer are carried out by adopting various natural plant compositions, such as a compound of medlar, radix rehmanniae, cortex lycii radicis, radix ophiopogonis, selfheal, nutmeg, moldavica dragonhead, subprostrate sophora, Chinese iris seed, corn stigma, dandelion, phellodendron, red paeony root, half of cattle, andrographis paniculata, fructus forsythiae, rhizoma atractylodis, mulberry leaf, deer medicine and Spongilla, a composition of mangnolia officinalis, rhizoma atractylodis, elecampane, rhizoma corydalis, concha arcae, oyster, bletilla striata and ginseng, and the like, and the extract of the formula is complex in proportioning and processing and expensive in price of certain components.

The Shibi tea is a wild plant, which is known as adinandrnitida (Adinandranitida), and is often eaten as a health-care tea and traditional Chinese herbal medicines for a long time. Research shows that the Shibi tea contains abundant flavonoid compounds (more than 20 percent), such as camellin A, camellin B and apigenin, wherein camellin A is mainly contained and accounts for about 15 percent. The Shibi tea extract has multiple curative effects of resisting oxidation, resisting cancer, easing pain, diminishing inflammation, reducing blood pressure and the like, and the main flavonoid camellia glycoside A of the Shibi tea extract has the effect of inhibiting the proliferation of liver cancer cells HepG2 and breast cancer cells MCF-7 and has stronger antioxidant activity. There is no relevant research on the therapeutic effect of Shibi tea and its main flavonoid camellin A in gastric ulcer.

Disclosure of Invention

In order to solve the problems, the invention provides a simple plant extract and application of rich active monomer components in the tea in a medicament for treating gastric ulcer, and discovers that the aqueous extract of the gecko tea and the active monomer camellin A thereof have good treatment effect on gastric ulcer through a mouse gastric ulcer model induced by hydrochloric acid alcohol, and the injury degree and inflammatory reaction of gastric ulcer are reduced by regulating NF kappa B inflammatory pathways and regulating the expression of related inflammatory factors IL-6.

The invention discloses the application value of the Shibi tea extract and the camellin A in the medicine for treating gastric ulcer for the first time.

The research theory of the invention is as follows: the pathogenesis of gastric ulcers is complex and multifactorial, with inflammatory reactions being one of the important mechanisms. Neutrophil infiltration in the damaged tissue induces the release of various inflammatory factors and enhances the inflammatory response of the gastric mucosa, and by inhibiting the inflammatory response, the exacerbation of gastric ulcers can be alleviated. The nuclear factor kappa B (NF-. kappa.B) signaling pathway plays an important role in the context of gastric mucositis. Under the drive of inflammatory factors and Reactive Oxygen Species (ROS), the activated NF-. kappa.B pathway exacerbates inflammation by triggering the transcription of inflammatory factors and chemokines, including tumor necrosis factor alpha (TNF-. alpha.) and interleukin 6 (IL-6). i κ B (inhibitor of NF- κ B) is an inhibitor of NF- κ B. Wherein in a resting state, two subunits of p65 and p50 of i kappa B-alpha and NF-kappa B exist in an inactivated state and are in cytoplasm. When an IKB kinase (IKK) is activated by an upstream signal, the activated IKK is ubiquitinated, phosphorylates and degrades i kappa B-alpha, so that two subunits of NF kappa B are activated from an inactivated state and transferred from cytoplasm to nucleus to be combined with corresponding inflammation-related genes, and the transcription of inflammatory factors such as TNF-alpha and IL-6 is started to induce inflammation. The invention discovers that the Shibi tea extract and the active monomer camellin A thereof can inhibit the degradation of i kappa B-alpha so as to inhibit NF kappa B signal channels, and can down-regulate the expression of inflammatory factors IL-6 so as to relieve the inflammatory reaction of gastric tissues.

The invention discovers that the Shibi tea extract and the active monomer camellin A thereof have the effect of treating acute gastric ulcer, and the principle of the extract is related to the inhibition of NF kappa B inflammatory pathways and the down-regulation of the expression of inflammatory factors.

Use of folium Photiniae and/or camellianin A in preparing medicine for treating gastric ulcer; or the application of the Shibi tea in preparing the medicine for treating gastric ulcer; or the application of the camellin A in preparing the medicine for treating the gastric ulcer is the important protection scope of the invention.

The camellin A is extracted from folium Claoxyli.

The Shibi tea is an aqueous extract of Shibi tea.

The preparation method of the aqueous extract of the Shibi tea comprises the following steps:

pulverizing dry folium Cladoniae Rangiferinae with pulverizer, extracting tea powder in boiling water for more than one time, filtering, mixing extractive solutions, concentrating the extractive solution by rotary evaporation, and freeze drying to obtain lyophilized powder of Cladonia Rangiferinae extract.

Preferably, the mass volume ratio of the Shibi tea to the water is 1: 15-25;

preferably, the extraction times are 2-6 times;

preferably, the time for each extraction is 25-35 min;

preferably, the extract is rotary-evaporated and concentrated at 50-70 ℃.

The mass-volume ratio of the stone wall tea to the water is 1: 20;

preferably, the number of extractions is 3;

preferably, the time of each extraction is 30 min;

preferably, the extract is concentrated by rotary evaporation at 60 ℃.

The purity of camellin A is greater than or equal to 98%;

preferably, the preparation method of camellin A is as follows: extracting 100g of Shibi tea with 1500mL of boiling water for 60min, filtering to obtain residue, extracting with 1200mL of boiling water for 60min, and filtering; mixing the two filtrates, storing at 2 deg.C for 24h, filtering, and collecting precipitate; drying the precipitate at 60 deg.C for 5h to obtain crude product; recrystallizing the crude product with 20% ethanol for 5 times, purifying to obtain crystal, and drying the crystal at 60 deg.C for 3 hr to obtain camellin A monomer.

The invention has the beneficial effects that after the Shibi tea extract and the active monomer component of the camelliaside A contained in the tea provided by the invention are applied to a medicament for treating gastric ulcer, through a mouse gastric ulcer model induced by hydrochloric acid alcohol, the Shibi tea aqueous extract and the active monomer camelliaside A thereof are found to have good treatment effect on gastric ulcer, and the damage degree and inflammatory reaction of gastric ulcer are reduced by regulating NF kappa B inflammatory pathways and regulating the expression of related inflammatory factors IL-6. Compared with the method mentioned in the background technology, the invention adopts simple raw materials, namely the aqueous extract of the stony wall tea and the active monomers with rich content in the tea, and the preparation method is relatively simple compared with the formula medicine.

Drawings

FIG. 1 is a schematic diagram of the technique of treating gastric ulcer with Shibi tea and camellin A;

FIG. 2 shows the therapeutic effect of Shibi tea and camellin A on hydrochloric alcohol induced gastric ulcer; (a) a stomach anatomy map; (b) an index of gastric injury;

FIG. 3HE staining observation stone wall tea and camellin A on stomach epithelium tissue recovery effect; (a) HE observation result graph; (b) an index of gastric injury;

FIG. 4 Sphaerotheca fuliginea and camellin A down-regulate the expression of IL-6; (a) IL-6 immunohistochemistry results; (b) IL-6 immunohistochemistry quantification results; (c) IL-6 Western Blot (WB) results; (d) IL-6 Western Blot (WB) quantification;

FIG. 5 Sphaerotheca littoralis and camellin A inhibit degradation of ikb-alpha; (a) (ii) ikb- α immunohistochemistry results; (b) quantitative results of i kappa B-alpha immunohistochemistry; (c) ikb- α Western Blot (WB) results; (d) quantitative results of i.kappa.B-. alpha.Western blot (WB).

Detailed Description

In order that those skilled in the art will better understand the present invention, the inventors will further describe and illustrate the present invention by the following specific examples, but do not limit the present invention.

Example 1

Preparation of Shibi tea freeze-dried powder and camellin A

Pulverizing dry folium Camelliae sinensis, and extracting in boiling water (the mass volume ratio of tea/water is 1: 20) for 30min for 3 times. Filtering, mixing the extractive solutions, steaming at 60 deg.C, concentrating, and freeze drying to obtain lyophilized powder.

Example 2

The present inventors have disclosed in earlier studies a method for producing camellin a: yuan, e.; liu, b.; ning, z.j.j.o.f.p.; preservation, Preparation and antioxidant activity of collagen A from Adinandritida leaves.2008,32(5),785 and 797. the method is as follows: extracting 100g of Shibi tea with 1500mL of boiling water for 60 min. The residue obtained by filtration was further extracted with 1200mL of boiling water for 60min and filtered. The two filtrates were mixed, stored at 2 ℃ for 24h, and then the precipitate was collected by filtration. The precipitate was dried at 60 ℃ for 5h to give the crude product. Recrystallizing the crude product with 20% ethanol for 5 times, and purifying to obtain crystal. Drying the crystal at 60 deg.C for 3h to obtain light yellow camellin A monomer with purity over 98%.

Example 3

1. Construction of acute gastritis animal model

Healthy 7-week-old male ICR mice were purchased, acclimated for 1 week, and then randomly grouped: (1) a normal control group; (2) a model control group; (3) a positive drug control group; (4) low dose camellin group A; (5) high dose camellin group A; (6) low dose litchis group; (7) high dose litchis groups of 7 animals each.

Except for normal control group, animals of each group are subjected to acute gastric ulcer molding injury by adopting 60% ethanol and 0.4M hydrochloric acid after fasting for one night, the gavage amount is 10mL/kg body weight, and the animals of the normal control group are subjected to gavage for one night and then are subjected to gavage with pure water in equal dosage as blank control. After the molding is successful, the drug administration treatment is carried out for 2 consecutive days every other day.

Cimetidine, camellianin A and SHIWANGCHA lyophilized powder are all dissolved in 0.5% CMCNa solution, and the intragastric volume of the animals is 10mL/kg body weight. Animals in the normal control group and model control group were gavaged with 0.5% CMCNa (solvent) as a control. The positive drug control group animals were gavaged with 100mg/kg cimetidine solution. Low-dose camellin A group animals are gavaged with 30mg/kg camellin A solution. High-dose camellin A group animals are gavaged with 100mg/kg camellin A solution. The low-dose Shibi tea group animals are gavaged with 200mg/kg Shibi tea freeze-dried powder solution. The high-dose Shibi tea group animals are gavaged with 700mg/kg Shibi tea freeze-dried powder solution.

After 2 days of intragastric administration, the animals were anesthetized and sacrificed 2h after the end of the last administration, and the stomach tissue was dissected.

2. Treatment and preservation of animal stomach tissue

The method comprises the steps of dissecting an animal to obtain stomach tissue, cutting the stomach wall along the greater curvature of the stomach, pouring out contents, slightly washing the inner wall of the stomach with pre-cooled normal saline, drying the inner wall of the stomach with a cotton ball, flattening the stomach in a small clean dish, placing the small dish containing the stomach tissue on grid paper, and taking a picture with a digital camera to record the damage condition of the inner wall of the stomach, so as to be convenient for later-stage stomach damage assessment. Subsequently, the stomach tissue is divided into two parts along the lesser curvature of the stomach, one half is frozen and stored at-80 ℃ for the subsequent protein level expression detection, and the other half is fixed in a neutral formaldehyde solution for the subsequent histopathological analysis and immunohistochemical analysis.

3. Anatomical analysis of gastric tissue

The gastric injury index statistics are carried out on the injury condition of the inner wall of the stomach of each animal in each group, and the statistics are as follows:

the length of erosion, ulcer, bleeding and the like limited to epithelial tissues on the inner wall of the stomach is taken as a cumulative integral, the normal score is 0, the spot erosion is counted for 1, the erosion length is counted for 2 when the length is less than 1mm, the erosion length is counted for 3 when the length is 1-2 mm, the erosion length is counted for 4 when the length is 3-4 mm, and the score is counted for 5 when the length is greater than 4mm, all the scores are added to be the total score of the damage index of the animal, and the average value of the damage.

4. Preparation of stomach tissue sections

(1) Tissue dehydration and Paraffin embedding

After the stomach tissue is soaked in neutral formaldehyde for 24 hours, the stomach tissue is taken out and placed in 70 percent ethanol for preservation. The stomach tissue is cut into pieces with a thickness of no more than 2mm with a sharp blade, placed in a tissue embedding cassette, and marked. The tissue is dehydrated according to the following steps: sequentially dehydrating in 70%, 80%, 90%, and 95% ethanol for 30min, and sequentially soaking in anhydrous ethanol (1; 2; 3) for 3 times, each for 20 min. Sequentially soaking in xylene solution (1; 2; 3) for 20min to make the tissue transparent. At present, the wax penetration is carried out for 40min by adopting 65 ℃ liquid soft wax, and then the wax penetration is carried out for 60min by adopting 65 ℃ hard wax. And taking out the tissue embedding box, and embedding the tissue by using a tissue embedding machine to enable the section of the tissue to be upward. The embedded tissue wax blocks were stored at-20 ℃.

(2) Tissue section

After the tissue wax block is refitted, the tissue wax block is sliced by a tissue slicer to a thickness of 4 mm. Spreading the slices on water at 42 deg.C, and baking the slices in an oven at 37 deg.C overnight. The prepared slices were stored in a refrigerator.

HE staining and histological analysis

After the tissue section is baked for 2 hours at 40 ℃, HE staining can be carried out. The method comprises the following steps: (1) dewaxing for 10min by using xylene 1; (2) dewaxing for 10min by using xylene 2; (3) rehydrating with anhydrous ethanol 1 for 5 min; (4) 2, rehydrating with absolute ethyl alcohol for 5 min; (5) 95% ethanol is rehydrated for 2 min; (6) rehydrating with 80% ethanol for 2 min; (7) 70% ethanol is rehydrated for 2 min; (8) washing with distilled water for 5 s; (9) staining with hematoxylin for 10 min; (10) soaking in 95% ethanol for 5 s; (11) washing with running water for 10 min; (12) washing with pure water for 5 s; (13) eosin staining for 40 s; (14) washing with distilled water for 10 s; (15) dehydrating with 95% ethanol for 2 min; (16) dehydrating with anhydrous ethanol for 2 min; (17) xylene 1 is transparent for 5 min; (18) xylene 2 is transparent for 5 min; (19) and (5) sealing the neutral gum. And (5) after the sealing is finished, naturally drying for 24 hours at normal temperature, and observing and analyzing by using an optical microscope.

The statistics of the tissue observation gastric injury index of the gastric epithelial tissue are as follows:

the results were scored for epithelial cell loss, mucosal sloughing and edema, hemorrhagic lesions, inflammatory cell infiltration, localized to epithelial tissue on the inside wall of the stomach. According to morphological observation results, the epithelial cell loss is counted by 0-3 points, the mucosal exfoliation and edema is counted by 0-4 points, the hemorrhagic injury is counted by 0-4 points, the inflammatory cell infiltration is counted by 0-3 points, all the points are added to be the total tissue morphological injury index of the animal, and the average value of the injury index of each group is calculated.

6. Immunohistochemical analysis

The tissue slices can be subjected to immunohistochemical experiments after being dried for 2 hours at 40 ℃. The method comprises the following specific steps:

(1) slice dewaxing and rehydration: dewaxing xylene 1 for 10min, dewaxing xylene 2 for 10min, rehydrating absolute ethyl alcohol 1 for 5min, rehydrating absolute ethyl alcohol 2 for 5min, rehydrating 95% ethyl alcohol for 2min, rehydrating 80% ethyl alcohol for 2min, and rehydrating 70% ethyl alcohol for 2 min;

(2) cleaning: soaking in pure water for 2min, and soaking in TBS for 5 min;

(3) soaking in 3% H at room temperature₂O₂Inactivating endogenous peroxidase for 15 min;

(4) washing with pure water for 1 time, and soaking in TBS for 5 min;

(5) repairing antigen: heating 1 × EDTA repair liquid 700ml in microwave oven (boiling EDTA repair liquid for 10-15 min: boiling in microwave oven with high fire for 8-10min, and then turning to middle and low fire for 10min), naturally cooling at room temperature for 30min

(6) Soaking and cleaning with TBS for 5min for 3 times;

(7) marking out a tissue area in the section by using an oil pen ring;

(8) dripping blocking solution (1 × TBST solution containing 5% serum) into the oil ring, and blocking at room temperature for 40 min;

(9) removing the sealing liquid with absorbent paper, adding primary antibody (diluted with sealing liquid) and incubating overnight in a wet box at 4 deg.C;

(10) recovering primary antibody after the to-be-sliced sheet is recovered to room temperature, and washing with TBS for 5min for 3 times;

(11) incubating with biotin-labeled secondary antibody at room temperature for 60min, and soaking and cleaning with TBS for 3 times, 5min each time;

(12) reacting with SABC reagent (HRP) at room temperature for 30min, and soaking in TBS for 3 times, 5min each time;

(13) DAB color development is carried out at room temperature, and the time of each group is not more than 5 min;

(14) washing with running water for 5min, and soaking in pure water for 1 min;

(15) counterstaining with hematoxylin for 1 min;

(16) washing with flowing water for 10min, dehydrating with 95% ethanol for 2min, dehydrating with 100% ethanol for 2min, and removing xylene twice, each for 5 min;

(17) neutral gum sealing sheet

And (5) after the sealing is finished, naturally drying for 24 hours at normal temperature, and observing and photographing by using an optical microscope. Immunohistochemical quantitative analysis was performed using ImageJ software.

7. Extraction and quantification of gastric tissue proteins

Extracting the gastric tissue protein:

taking 20mg of each group of gastric tissues, adding 0.2ml of RIPA cell lysate (containing 1% PMSF), fully homogenizing on ice in a homogenizer, and collecting homogenate in a 1.5ml EP tube; the homogenate was lysed on ice for 1h and centrifuged at 13200r/min at 4 ℃ for 5min, the supernatant was taken (5. mu.L of each was used separately for protein content determination), and the remainder was stored at-20 ℃.

And (3) protein quantification:

(1) preparing a BCA working solution: uniformly mixing the solution A and the solution B according to the required amount for measurement in a ratio of 50: 1;

(2) preparing a protein standard solution: preparing 0, 0.125, 0.25, 0.5, 1.0 and 2.0mg/mL protein standard substance;

(3) adopting a 96-hole enzyme label plate, firstly adding 25 times diluted sample (1 mu L sample +24 mu L diluent) and 25 mu L standard sample, then adding 200 mu L working solution, setting three parallel repeat, and tapping and mixing uniformly;

(4) after incubation for 30min at constant temperature of 37 ℃, an OD value is measured by an enzyme-labeling instrument at the wavelength of 560nm, and the protein concentration of each sample is calculated according to a standard curve.

Western blotting detection of protein level expression

(1) Preparing 10% SDA-PAGE gel;

(2) dissolving the gastric tissue protein, and adjusting the concentration by adopting a protein lysate and 4 multiplied by protein gel electrophoresis loading buffer solution according to the quantitative concentration of the protein so as to ensure that the final concentration of the sample protein is 25 mu g/mu L. Heating and denaturing the tissue protein liquid with the adjusted concentration at 98 ℃ for 10min, immediately inserting the tissue protein liquid into ice for cooling, and carrying out short-time vortex centrifugation on the sample to obtain a sample;

(3) pouring the electrophoresis buffer solution into an electrophoresis tank to a proper liquid level, and inserting the electrophoresis buffer solution into a gel plate. Slightly pulling out the comb, sucking out the denatured protein liquid, tightly attaching the denatured protein liquid to the upper part of the sample adding hole, and slightly adding the sample into the gel hole; the electrophoresis apparatus is set to a voltage-stabilizing state, the power supply is switched on, and the voltage is adjusted to 80V so that the sample passes through the concentrated gel. After the dye enters the separation gel, the voltage is increased to 120V, and electrophoresis is continued to enable the dye to reach a proper position of the separation gel;

(4) stopping electrophoresis after protein separation is complete, carefully taking out gel, removing a concentrated gel part, preparing a PVDF membrane with proper size in advance, soaking in methanol for about 5min, and sequentially arranging an anode plate, a sponge, filter paper, gel, the PVDF membrane, the filter paper, the sponge and a cathode plate on a membrane rotating device from bottom to top; ensuring good contact among all layers, and discharging bubbles on the contact surface of each layer of filter paper;

(5) and inserting the prepared film transferring clamp into a film transferring groove, adding a film transferring liquid precooled at 4 ℃ in advance, and correctly connecting a power supply to ensure that charges flow from a negative electrode to a positive electrode. Placing in a box filled with ice, switching on a power supply, and operating at constant current 275mA for 60-90 min;

(6) carefully taking out the transfer membrane after the membrane transfer is finished, marking the upper right corner of the membrane, rinsing the membrane in 1 xTBST solution once, putting the membrane into sealing solution containing 5% of skimmed milk powder, and slowly shaking the membrane on a shaking table at room temperature for sealing for 1-2 h;

(7) cutting a PVDF membrane according to the molecular weight of the protein, adding a primary antibody diluted by a confining liquid according to a certain proportion, and incubating overnight at 4 ℃;

(8) recovering primary antibody, rinsing with 1 × TBST for 3 times, each time for 5min, adding secondary antibody diluted with 1 × TBST according to a certain proportion, incubating for 50min, recovering secondary antibody, and washing membrane with 1 × TBST for 3 times, each time for 5 min;

(9) according to the following steps: 1(v/v) mixing two liquids in the ECL kit, uniformly spreading the mixed liquid on the surface of the PVDF membrane, and acting for 2min at room temperature. The film was developed in a Canon gel imaging system.

As can be seen from the attached figure 2, the hydrochloric acid alcohol modeling causes burning injury to the inner wall of the stomach of the mouse and red blood streak. The Shibi tea and camellin A treatment group has treatment and recovery effects on the gastric ulcer induced by the hydrochloric acid alcohol, and the effect is superior to that of the positive drug treatment group. Control: normal control group, Model: model control, Positive: cimetidine (100mg/kg), L-CA: low concentration camellin a (30mg/kg b.w.), H-CA: high concentration camellin a (100mg/kg b.w.), L-Tea: low-concentration freeze-dried Shibi tea powder (200mg/kg b.w.), H-CA: high concentration freeze-dried Shibi tea powder (700mg/kg b.w.), all values are expressed as mean + -SD, # # P <0.01 compared with Control; p <0.01 compared to Model.

As can be seen from the attached figure 3, the HE staining result shows that the epithelial cells of the gastric mucosa of the normal group are complete, the arrangement of the gland bodies in the mucosa is regular and compact, and the phenomena of inflammatory cell infiltration and epithelial exfoliation are not obvious; the epithelial cells of the gastric mucosa of the model group are loosely arranged and necrotized, and have obvious infiltration of a plurality of inflammatory cells and the epithelial exfoliation phenomenon; the yang medicine group has complete cells, the gland cells are arranged regularly, and no obvious inflammatory cell infiltration exists; epithelial cells of the L-CA group and the L-Tea group are complete and are arranged regularly, a mucous layer slightly falls off, and no obvious inflammatory cell infiltration exists; the epithelial cells of the H-CA group and the H-Tea group are complete and orderly arranged, and the submucosa has no obvious inflammatory cell infiltration and no obvious epithelial exfoliation phenomenon. Control: normal control group, Model: model control, Positive: cimetidine (100mg/kg), L-CA: low concentration camellin a (30mg/kg b.w.), H-CA: high concentration camellin a (100mg/kg b.w.), L-Tea: low-concentration freeze-dried Shibi tea powder (200mg/kg b.w.), H-CA: high concentration freeze-dried Shibi tea powder (700mg/kg b.w.), all values are expressed as mean + -SD, # # P <0.01 compared with Control; p <0.01 compared to Model.

As can be seen from FIG. 4, the immunohistochemistry and WB results indicate that Shiparicha and camellin A have the same effect of inhibiting the expression of the inflammatory factor IL6 as the positive drugs. Control: normal control group, Model: model control, Positive: cimetidine (100mg/kg), L-CA: low concentration camellin A

(30mg/kg b.w.), H-CA: high concentration camellin a (100mg/kg b.w.), L-Tea: low-concentration freeze-dried Shibi tea powder (200mg/kg b.w.), H-CA: high concentration freeze-dried Shibi tea powder (700mg/kg b.w.), all values are expressed as mean + -SD, # # P <0.01 compared with Control; p <0.01 compared to Model.

As can be seen from FIG. 5, i κ B- α is an upstream factor of NF κ B, binds to NF κ B and inhibits its activity, and immunohistochemistry and WB results indicate that the total i κ B- α expression of the model group is reduced, indicating that i κ B- α is activated, i κ B- α activation releases NF κ B, and NF κ B inflammatory pathways are activated; the gecko tea and camellin A treatment group can inhibit the degradation of i kappa B-alpha in a gradient manner, which indicates that the inflammatory pathway of NF kappa B can be inhibited. Control: normal control group, Model: model control, Positive: cimetidine (100mg/kg), L-CA: low concentration camellin a (30mg/kg b.w.), H-CA: high concentration camellin a (100mg/kg b.w.), L-Tea: low-concentration freeze-dried Shibi tea powder (200mg/kg b.w.), H-CA: high concentration freeze-dried Shibi tea powder (700mg/kg b.w.), all values are expressed as mean + -SD, # # P <0.01 compared with Control; p <0.01 compared to Model.

The invention creatively selects the Shibi tea as the raw material and is applied to the preparation of the medicine for treating gastric ulcer;

the active monomer camellin A extracted from the Shibi tea is rich in the tea, and can be directly extracted and separated from the tea leaves or purchased;

the tea aqueous extract and the monomer have good effect of treating gastric ulcer in the research concentration, and have no toxicity to animals and high safety.

Through the experiments, the Shibi tea extract and the camellin A are proved to be capable of inhibiting NF kappa B inflammatory pathways by regulating the activity of i kappa B-alpha, down-regulating the expression of an inflammatory factor IL-6 and relieving the gastric mucositis reaction; the Shibi tea extract and the camellin A have good treatment effect on the gastric ulcer of mice induced by hydrochloric acid alcohol, reduce the injury area and the injury index of gastric mucosa, and reduce the inflammatory infiltration of gastric epithelial tissues.

Claims

1. Use of folium Photiniae or camellianin A in preparing medicine for treating gastric ulcer is provided.

2. The use according to claim 1, wherein the camellin A is obtained by extraction from Shimantha tea.

3. The use of claim 1, wherein the Shibi tea is an aqueous extract of Shibi tea.

4. Use according to claim 3, wherein the aqueous extract of Shibi tea is prepared by:

pulverizing dry folium Cladoniae Rangiferinae, extracting the obtained tea powder in boiling water for at least one time, filtering, mixing the extractive solutions, concentrating the extractive solution by rotary evaporation, and freeze drying to obtain lyophilized powder of Cladonia Rangiferinae extract.

5. The use as claimed in claim 4, wherein the mass to volume ratio of Shibi tea to water is from 1: 15 to 25.

6. The use according to claim 5, wherein the number of extractions is 2 to 6.

7. The use according to claim 6, wherein the time for each extraction is 25-35 min; and (3) carrying out rotary evaporation and concentration on the extracting solution at 50-70 ℃.

8. The use as claimed in claim 4, wherein the mass to volume ratio of Shibi tea to water is from 1: 20; the extraction times are 3 times; the extraction time is 30 min; the extract was concentrated by rotary evaporation at 60 ℃.

9. Use according to claim 2, wherein the camellin A is greater than or equal to 98% pure.

10. Use according to claim 9, wherein camellin a is prepared by the method comprising: extracting 100g of Shibi tea with 1500mL of boiling water for 60min, filtering, extracting the residue with 1200mL of boiling water for 60min, and filtering; mixing the two filtrates, storing at 2 deg.C for 24h, filtering, and collecting precipitate; drying the precipitate at 60 deg.C for 5h to obtain crude product; recrystallizing the crude product with 20% ethanol for 5 times, purifying to obtain crystal, and drying the crystal at 60 deg.C for 3 hr to obtain camellin A monomer.