Application of Chinese knotweed rhizome and aloe bowel-relaxing composition in preparation of anti-helicobacter pylori drugs

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a novel application of a Chinese knotweed rhizome and aloe bowel-relaxing composition in preparation of a medicine for preventing or treating helicobacter pylori infection. The fallopia multiflora and aloe bowel-relaxing composition consists of fallopia multiflora, semen cassiae, immature bitter orange, aloe, donkey-hide gelatin, medlar, bighead atractylodes rhizome and ginseng. Experiments show that the Chinese knotweed root-aloe bowel-relaxing composition has relatively strong bacteriostatic and bactericidal effects on helicobacter pylori, not only influences the micromorphology and cell adhesion ability of the helicobacter pylori thalli, but also influences the genes of the helicobacter pylori thalli.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology and relates to a new use of a Shouhui laxative composition, and specifically relates to a new use of the Shouhui laxative composition in preparing a drug for preventing or treating Helicobacter pylori infection. Background Art

[0002] Helicobacter pylori (HP) is a Gram-negative, microaerophilic, spiral-shaped bacterium that mainly parasitizes the human gastric mucosa and upper duodenum. It is the only known bacterium that can survive in the highly acidic environment of the stomach. Helicobacter pylori is spiral or arc-shaped, with a diameter of about 0.5 microns and a length of 2.5 to 4 microns. It has 4-6 flagella that enable it to move under the gastric mucosa. Helicobacter pylori is adapted to grow in a low-oxygen environment, which enables it to survive in the hypoxic environment under the gastric mucosa. It decomposes urea into ammonia and carbon dioxide through urease, thereby neutralizing gastric acid, which is also an important mechanism for its survival in the stomach. Helicobacter pylori is mainly transmitted through oral-oral and fecal-oral routes, which are common in shared meals, close contact and poor hygiene habits. Helicobacter pylori infection is closely related to a variety of gastric diseases, including chronic gastritis, gastric ulcers, duodenal ulcers and gastric cancer. It is the main pathogenic factor of gastric ulcer and duodenal ulcer. Helicobacter pylori can be detected in about 80% of gastric ulcer patients and more than 90% of duodenal ulcer patients.

[0003] Helicobacter pylori is classified as a Class I carcinogen by the World Health Organization (WHO) and is an important factor leading to gastric cancer. Long-term infection can cause chronic inflammation of the gastric mucosa, gradually developing into atrophic gastritis, intestinal metaplasia, dysplasia, and may eventually develop into gastric cancer. In addition, Helicobacter pylori infection may also be associated with other diseases, such as gastric MALT lymphoma, idiopathic thrombocytopenic purpura, and some skin diseases. Common diagnostic methods include 13C or 14C breath test, serum antibody test, stool antigen test, rapid urease test under gastroscopy, and tissue biopsy. The standard treatment for Helicobacter pylori infection is the combination of antibiotics and proton pump inhibitors (PPIs), with common regimens being triple therapy (two antibiotics plus one PPI) or quadruple therapy (two antibiotics plus one PPI and one bismuth agent). With the widespread use of antibiotics, Helicobacter pylori has significantly increased its resistance to a variety of commonly used antibiotics (such as clarithromycin, metronidazole, and levofloxacin). This has led to a decrease in eradication rates and an increased risk of treatment failure. At the same time, antibiotics not only kill Helicobacter pylori, but may also damage the normal flora in the intestines, leading to dysbacteriosis and increasing the chance of infection.

[0004] In recent years, there have been many reports on medicines or health products for treating or preventing Helicobacter pylori infection: CN118593640A discloses a Chinese medicine composition for resisting Helicobacter pylori, comprising Rhizoma Coptidis, processed Magnolia officinalis, Rhizoma Phragmites, processed Pinellia tuber, Fructus Douchi, Rhizoma Acorus, Fructus Gardeniae and Herba Taraxaci, and the composition is made into a mouthwash; CN110354010A discloses a mouthwash with anti-Helicobacter pylori efficacy, comprising A Chinese medicine active component, B filler, C flavoring agent and D solvent, wherein A Chinese medicine active component comprises Rhein extract, Berberine extract, Matrine extract, Baicalin extract and Mango Ginger Joint Phenol extract. The existing technology or marketed products have the disadvantages of unclear efficacy and weak pertinence.

[0005] CN100453105C discloses a Shouhui laxative capsule, which is made of Polygonum multiflorum, aloe, Cassia seed, wolfberry, donkey-hide gelatin, ginseng, Atractylodes macrocephala, and Citrus aurantium. It obtained the production approval (National Medicine Standard Z20150041) and the new drug certificate (National Medicine Certificate Z20150005) on May 6, 2015, and was launched on February 16, 2020. The trade name is Shouhui Tongbian Capsule. It is an exclusive product of Lunan Houpu Pharmaceutical Co., Ltd. Its prescription is derived from the clinical prescription of famous old Chinese medicine practitioners. It is mainly used clinically for the treatment of functional constipation. It has the effects of moisturizing the intestines and promoting bowel movements, purging turbidity and detoxifying, nourishing blood and moisturizing dryness, and nourishing qi and strengthening the spleen. It has a good moisturizing and laxative therapeutic effect on constipation caused by internal accumulation of toxic evil and deficiency of yin fluid, can comprehensively improve gastrointestinal function, has a definite effect, fast onset, no toxic side effects, and has the effects of detoxification, beauty, weight loss and lipid reduction. It is especially suitable for the elderly and those with weak constitutions. In recent years, with the in-depth research on Chinese medicine preparations, more and more new effects of Shouhui laxative preparations have been discovered.

[0006] Currently, there is no research or report on the use of Shouhui Tongbian Capsule in the treatment or prevention of Helicobacter pylori infection. Summary of the invention

[0007] In view of the defects in the prior art for the prevention and treatment of Helicobacter pylori infection, the present invention develops new uses based on the company's patent CN100453105C, the invention name of which is "a composition with laxative, detoxifying, weight loss and lipid-lowering functions and a preparation method", specifically a new use in the preparation of drugs for the prevention or treatment of Helicobacter pylori infection.

[0008] The Shouhui laxative Chinese medicine composition of the invention is composed of Polygonum multiflorum, Cassia seed, Citrus aurantium, aloe, donkey-hide gelatin, wolfberry, Atractylodes macrocephala and ginseng.

[0009] The components in the composition of the present invention are prepared according to the preparation or extraction methods of the components in CN100453105C, a composition with laxative, detoxifying, weight-loss and lipid-lowering functions and its preparation method.

[0010] The components in the composition are calculated by weight as follows: 60-150 parts by weight of Polygonum multiflorum, 80-180 parts by weight of Cassia seed, 50-200 parts by weight of Citrus aurantium, 100-200 parts by weight of Aloe, 30-150 parts by weight of Donkey-hide gelatin, 30-150 parts by weight of Lycium barbarum, 20-100 parts by weight of Atractylodes macrocephala, and 20-100 parts by weight of Ginseng; preferably, the composition comprises 120 parts by weight of Polygonum multiflorum, 140 parts by weight of Cassia seed, 120 parts by weight of Citrus aurantium, 160 parts by weight of Aloe, 75 parts by weight of Donkey-hide gelatin, 75 parts by weight of Lycium barbarum, 50 parts by weight of Atractylodes macrocephala, and 50 parts by weight of Ginseng.

[0011] The present invention protects the novel use of the above-mentioned Chinese medicine composition in the preparation of a drug for treating or preventing Helicobacter pylori infection. The composition can significantly inhibit Helicobacter pylori, has a lower effective antibacterial concentration than the antibiotics in the prior art, and can also prevent or treat diseases such as gastric ulcers caused by Helicobacter pylori.

[0012] Preferably, the use includes type I Helicobacter pylori and type II Helicobacter pylori infection, the type I Helicobacter pylori infection can cause diseases such as gastric ulcer and duodenal ulcer, and the type II Helicobacter pylori infection can cause diseases such as chronic gastritis.

[0013] Preferably, in the above use, the composition can be used in combination with antibiotics to prevent or treat Helicobacter pylori infection.

[0014] Further preferably, the antibiotic is selected from one or more of clarithromycin, amoxicillin, levofloxacin or metronidazole.

[0015] The composition of the present invention can be used to prepare medicines, health products and foods, including but not limited to medicines.

[0016] The Chinese medicine composition provided by the present invention may also contain pharmaceutically acceptable excipients and be prepared into a pharmaceutical preparation according to conventional preparation processes.

[0017] Preferably, in the use, the composition is a capsule, granule, pill or tablet; further preferably, the composition is Shouhui Tongbian Capsule.

[0018] The invention object of the present invention is mainly achieved through the following technical solutions:

[0019] Example 1 shows that the Shouhui laxative composition has strong antibacterial and bactericidal effects on Helicobacter pylori.

[0020] Example 2 is a dynamic investigation of the strong antibacterial effect of the Shouhui laxative composition on Helicobacter pylori.

[0021] Example 3 shows that the Shouhui laxative composition affects the microscopic morphology of Helicobacter pylori cells, causing damage to the head and tail of Hp.

[0022] Example 4 shows that the Shouhui laxative composition can significantly reduce the ability of Helicobacter pylori to adhere to cells.

[0023] Example 5 shows that the Shouhui laxative composition affects the genes of Helicobacter pylori.

[0024] Example 6 shows that the Shouhui laxative composition can be used in combination with antibiotics such as clarithromycin, amoxicillin, levofloxacin, metronidazole, etc., without antagonism, which is helpful to solve the problem of antibiotic resistance.

[0025] In the composition of the present invention, eight Chinese medicinal herbs are used in combination to produce a significant synergistic effect, thereby significantly improving the effect of preventing and treating Helicobacter pylori infection.

[0026] The inventors found in the study that eight kinds of Chinese medicine were extracted separately by continuous reflux of 75% ethanol, and the ATCC 700392MIC was used to detect the inhibitory effect of various Chinese medicines on Helicobacter pylori. The results showed that the minimum inhibitory concentration of aloe vera was 640μg / mL, cassia seed was 1280μg / mL, and the remaining Chinese medicines were greater than 2580μg / mL. After using the Shouhui laxative preparation, the inhibitory concentration was significantly reduced.

[0027] Further studies have shown that in the Chinese medicine composition of the present invention, Polygonum multiflorum is bitter, sweet, astringent, slightly warm in nature, and enters the liver and kidney meridians, and has the effects of nourishing the liver and kidney, replenishing essence and blood, blackening hair, strengthening tendons and bones, and moistening the intestines and relieving constipation. Rhein in Polygonum multiflorum can inhibit the proliferation and glucose consumption of human gastric adenocarcinoma BGC-823 cells, and can also cause ROS accumulation in HepG2 cells, causing cell mitochondrial damage, thereby accelerating cell apoptosis. Another study found that the ethyl acetate extract of Polygonum multiflorum has a blocking effect on G2 phase cells, can induce cell apoptosis in vitro, and can inhibit tumor growth and angiogenesis in vivo, and has a broad-spectrum anti-cancer activity.

[0028] In the Chinese medicine composition of the present invention, Cassia seed is bitter, sweet and slightly cold, enters the liver and kidney, and has the effects of clearing the liver and improving eyesight, promoting diuresis and relieving constipation. The main components of Cassia seed include chrysophanol, rhein, rhein, etc. These anthraquinone compounds have a strong inhibitory effect on the growth of Helicobacter pylori.

[0029] In the Chinese medicine composition of the present invention, the bitter, pungent, sour and slightly cold nature of the immature bitter fruit enters the spleen and stomach meridians, and has the effects of breaking up qi and eliminating accumulation, resolving phlegm and dispersing lumps. It can treat gastrointestinal qi stagnation and has the function of bidirectionally regulating gastrointestinal smooth muscle. Studies have found that the Zhishi Xiaopi Decoction can significantly increase the patient's pepsinogen and serum gastrin levels in the treatment of HP-related gastritis patients, thereby inhibiting the inflammatory reaction of the gastric mucosa, thereby improving symptoms and improving the efficacy.

[0030] In the Chinese medicine composition of the present invention, aloe vera is cold in nature and bitter in taste, and enters the liver and large intestine meridians, and has the functions of purgation, liver cleansing, and insecticidal. Natural antibacterial substances in aloe vera, such as aloe-emodin, aloin, and aloe polysaccharides, can interfere with the cell wall structure of Helicobacter pylori and inhibit its growth and reproduction. The inventors also found that aloe vera has a strong anti-inflammatory effect, and can reduce the excessive gastric acid secretion caused by Helicobacter pylori by reducing the inflammatory reaction of the gastric mucosa; the antioxidant substances contained in aloe vera can reduce the oxidative stress caused by Helicobacter pylori infection, reduce the damage of gastric mucosal cells, and alleviate symptoms such as gastritis and ulcers.

[0031] In the traditional Chinese medicine composition of the present invention, donkey-hide gelatin is mild in nature and sweet in taste, and has the effects of nourishing blood and yin. Its unique polyanion base structure increases the resistance to bacterial perforation in immunity, infection and inflammatory reactions, forming a barrier, thereby forming a strong protective effect on the gastric mucosa and promoting the rapid healing of the gastric mucosa of patients infected with Helicobacter pylori.

[0032] In the traditional Chinese medicine composition of the present invention, Atractylodes macrocephala can remove oxygen free radicals in the body and regulate the immune function of the body.

[0033] In the Chinese medicine composition of the present invention, ginseng tastes sweet and slightly bitter, is slightly warm in nature, and enters the spleen, lung, heart, and kidney meridians. It has the effects of greatly replenishing vital energy, restoring pulse and strengthening deficiency, replenishing spleen and lungs, promoting body fluid and nourishing blood, and calming the nerves and improving intelligence. The acidic polysaccharide in ginseng inhibits the adhesion of Helicobacter pylori to human gastric adenocarcinoma epithelial cells.

[0034] In the Chinese medicine composition of the present invention, the combined use of Polygonum multiflorum, Cassia seed, Citrus aurantium, aloe, donkey-hide gelatin, Chinese wolfberry, Atractylodes macrocephala and ginseng has a significantly better effect of inhibiting Helicobacter pylori than any single Chinese medicine. The combination of the medicines has a slow attack, strengthens the body and eliminates evil, inhibits the growth of Helicobacter pylori in the body, and achieves the purpose of clearing turbid air in the body.

[0035] Compared with the prior art, the present invention has the following outstanding advantages:

[0036] The present invention provides a new use of the Shouhui laxative composition for resisting Helicobacter pylori, and experimentally verifies that it has a significant inhibitory effect on Helicobacter pylori. The drug combination in the prescription may achieve the purpose of comprehensive conditioning of the body by regulating the functions of human internal organs, the circulation of qi and blood, etc., rather than just treating a single symptom. Traditional Chinese medicine can comprehensively improve symptoms and improve the quality of life of patients. Compared with antibiotics, the side effects of traditional Chinese medicine are relatively mild and have less overall impact on the body. Moreover, Shouhui laxative capsules have the effects of warming the middle and removing phlegm, resolving phlegm and stopping vomiting, etc., which can promote the coordinated operation of organs in the body and enhance human immunity. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 This is a diagram showing the dynamic investigation of the antibacterial effect of the Shouhui laxative composition on Helicobacter pylori in Example 2.

[0038] Figure 2 This is a diagram showing the effect of the drug administration group in Example 3 on the microscopic morphology of Helicobacter pylori cells.

[0039] Figure 3 The figure shows the microscopic morphology of Helicobacter pylori cells in the growth control group in Example 3.

[0040] Figure 4 This is a graph showing the effect of the Shouhui laxative composition at different concentrations in Example 4 on the ability of Helicobacter pylori to adhere to cells.

[0041] Figure 5 This is a diagram showing the effect of the Shouhui laxative composition in Example 5 on the Helicobacter pylori gene. DETAILED DESCRIPTION

[0042] The following specific implementation modes are listed to further illustrate the present invention, but do not limit the scope of the present invention in any way. Those skilled in the art can make various modifications or improvements based on the basic idea of ​​the present invention, but as long as they do not deviate from the basic idea of ​​the present invention, they are all within the scope of the present invention.

[0043] The Shouhui laxative composition used in the following examples of the present invention is Shouhui laxative capsules, produced by Lunan Houpu Pharmaceutical Co., Ltd., batch number: 141423023, and the positive control drug is clarithromycin (Shanghai McLean Biochemical Co., Ltd., China). Strains: Helicobacter pylori standard strain ATCC700392, clinically isolated Helicobacter pylori strains SS1, CS01, QYZ-001, QYZ-004.

[0044] Example 1: Investigation of the effect of the Shouhui laxative composition on inhibiting or killing Helicobacter pylori

[0045] Method for determining the minimum inhibitory concentration (MIC): weigh 10.24 mg of Shouhui Tongbian Capsule (without the capsule shell), add 2 mL of brain heart infusion to dissolve and mix, centrifuge at 12000 rpm for 3 min, filter the supernatant through a 0.22 μm organic filter membrane, and obtain a 5.12 mg / mL stock solution. Take 500 μL of the stock solution + 500 μL of brain heart infusion to prepare multiple concentration gradients of 2560, 1280, 640, 320, 160, 80 μg / mL by double dilution method, dissolve the antibiotic in dimethyl sulfoxide (DMSO) to prepare a stock solution, dilute it to multiple concentration gradients (DMSO content ≤ 1%) when used, draw 50 μL of the prepared drug solution and add it to a 96-well plate, set at least 3 duplicate wells for each drug concentration, set a negative control well (containing only the drug, not inoculated with the strain), and a growth control well (containing no drug, only inoculated with the strain). Take out the strain from the incubator, scrape the bacterial moss with a moistened sterile cotton swab into the brain heart infusion, adjust the turbidity of the bacterial suspension to 1McF with brain heart infusion containing 20% ​​FBS, dilute 10 times, inoculate 50μL per well in the above-mentioned 96-well plate containing the drug (the final turbidity of the bacterial suspension in the 96-well plate is about 1×106CFU / mL), place the 96-well plate in the incubator, and culture it under microaerobic conditions at 37°C and 150rpm for 72 hours before taking out and observing the results. The results are available only when the growth control appears button-shaped or obviously turbid. The clearest well observed by naked eye is the lowest drug concentration, which is the MIC (minimum inhibitory concentration). Clarithromycin was used as a positive control in each experiment, and the MIC value determination was repeated 3 times. The results are shown in Table 1 below.

[0046] Method for determining the minimum bactericidal concentration (MBC): Take out the 96-well plate incubated for 72 hours in the microbroth dilution method to determine the MIC, prepare multiple concentration gradients of Shouhui Tongbian Capsules such as MIC, 2MIC, and 4MIC, and then draw 100μL of the liquid in the MIC, 2MIC, and 4MIC drug concentration wells and evenly spread them on the fresh blood plate. After culturing for 72 hours, take out the observation results to reduce the initial inoculation concentration by 10 3 The lowest drug concentration at CFU / mL is MBC. The results are shown in Table 1 below.

[0047] Table 1

[0048] Helicobacter pylori strains Drug sensitivity MIC (μg / mL) MBC (μg / mL) ATCC 700392 S 320 640 SS1 S 320 640 CS01 R(CLR) 320 640 QYZ-001 R(MTZ) 160 320 QYZ-004 R (CLR, MTZ, LEF, AMO) 160 320

[0049] Note: S stands for drug sensitivity, R stands for drug resistance, CLR stands for clarithromycin, MTZ stands for metronidazole, LEF stands for levofloxacin, and AMO stands for amoxicillin.

[0050] The results showed that the Shouhui laxative composition had a dual effect of antibacterial and bactericidal effects on Helicobacter pylori; in addition, the Shouhui laxative composition also had a strong inhibitory effect on Helicobacter pylori isolated clinically, indicating that the Shouhui laxative composition can solve the problem of antibiotic resistance to a certain extent.

[0051] Example 2: Dynamic investigation of the antibacterial effect of the Shouhui laxative composition on Helicobacter pylori

[0052] Determination method: Prepare Shouhui Tongbian Capsules (without capsule shell) with multiple concentration gradients such as MIC, 1 / 2MIC, and 1 / 4MIC by the two-fold dilution method, and use the brain heart extract without medicine as the growth control group. Taking ATCC 700392 as a representative, adjust the turbidity of the bacterial suspension as described in Example 1, with a ratio of bacterial suspension: drug solution / brain heart extract without medicine of 1:1, inoculate 2mL per well in a 6-well plate, place in a three-gas incubator, 37°C, microaerobic environment, 150rpm shaking culture, aspirate 100μL of culture solution in a 96-well plate at 0, 8, 12, 24, 28, 36, 48, 60, and 72h to measure OD 600nm. The larger the absorbance value, the more bacteria there are and the worse the antibacterial effect of the drug. Repeat the experiment 3 times and take the average value. The experimental results are as follows Figure 1 shown.

[0053] The results showed that the Shouhui laxative composition had a good antibacterial effect on Helicobacter pylori at MIC ~ 1 / 4MIC, and could inhibit the growth of Helicobacter pylori within 72 hours. The effect of inhibiting Helicobacter pylori showed a dose-increasing trend.

[0054] Example 3: Investigating the effect of the Shouhui laxative composition on the microscopic morphology of Helicobacter pylori cells

[0055] Determination method: Use a sterile cotton swab to scrape the mature ATCC 700392 strain grown on the blood plate, adjust its turbidity to 1 MCF, culture it in 10% FBS BHI for 24 hours, then take 1 mL of the bacterial solution and put it in 49 mL 10% FBS BHI, mix it, and treat the ATCC 700392 strain with or without MIC drug concentration for 12 hours, collect the bacteria by centrifugation at 5000 rpm for 3 minutes, wash it with PBS three times, fix it at 4°C overnight with electron microscopy fixative, and use a graded ethanol series to initially dehydrate the sample, then freeze-dry and fix it. The sample was sprayed with gold and observed by scanning electron microscope (SU8020, Hitachi, Japan). The experimental results are as follows. Figure 2 and 3 As shown, Figure 2 The left and right pictures are respectively the effects of the drug administration group on the microscopic morphology of Helicobacter pylori under microscopes of different magnifications; Figure 3The left and right pictures show the microscopic morphology of Helicobacter pylori in the growth control group under microscopes of different magnifications.

[0056] The electron microscope image showed the ultrastructure of Hp. The surface of the bacteria in the growth control group was smooth and intact, and most of them were in the shape of spiral rods. The head and tail of Hp in the group treated with the Shouhui laxative composition were damaged, which affected the survival of Helicobacter pylori.

[0057] Example 4: Investigating the effect of the Shouhui laxative composition on the ability of Helicobacter pylori to adhere to cells

[0058] Assay method: GES-1 cells (4×10 5 / well) were inoculated in a 6-well plate for 24 hours, and then treated with different concentrations of Shouhui Tongbian Capsules (without the capsule shell) (0, 160, 320, 640 μg / mL). 4 hours later, infection was carried out with 1% FITC-stained ATCC 700392 (MOI of 100) for 8 hours. After removing the non-adherent Helicobacter pylori, fix it with 4% paraformaldehyde fixative for 20 minutes. Subsequently, the 4% paraformaldehyde fixative was discarded, and DAPI staining solution and anti-fluorescence quencher were added. Finally, the fluorescence signal was analyzed using an IM-5FLD fluorescence inverted microscope. The experimental results are shown in Figure 4 shown.

[0059] Figure 4 In the figure, blue fluorescence represents GES-1 cells, and green fluorescence represents the number of Helicobacter pylori. After administration of the Shouhui laxative composition, the adhesion of Helicobacter pylori to GES-1 cells was significantly reduced, and as the administration concentration increased, the adhesion ability was gradually inhibited.

[0060] Example 5: Using RT-qPCR analysis method to investigate the effect of the Shouhui laxative composition on Helicobacter pylori genes

[0061] Determination method: In order to collect enough bacteria, Helicobacter pylori ATCC 700392 was adjusted to 1 MCF using a DensiCHEK Plus turbidimeter and cultured for 2 days. 1 mL of bacterial solution (OD 600 = 0.5-0.6) was diluted 50 times in BHI broth and 10% FBS (with or without MIC of Shouhui Tongbian Capsules (capsule shell removed)) was added for 24 hours. The total RNA of the sample was extracted using the Purelink RNA kit. Its concentration was determined using a NanoDrop 2000 spectrophotometer (Thermo Scientific, USA). On the Thermo Scientific 7500 Fast Real-Time PCR System (Real-Time Quantitative PCR System), SYBR PremixEx Tap TMKit (Takara) was used and RT-qPCR was performed according to the manufacturer's instructions. The data were verified using the 2-ΔΔCt method. Table 2 lists the specific primers used to amplify mRNA fragments. After normalization with the internal reference gene 16S, the expression regulation of these genes was investigated by comparison with the control bacteria. The experimental results are shown in Figure 5 shown.

[0062] Table 2 Primers used for RT-qPCR analysis

[0063]

[0064]

[0065] Figure 5 In the experiment, Helicobacter pylori strain ATCC 00392 was used, the concentration of Shouhui Tongbian Capsule was 320 μg / mL, and the incubation time was 24 h. * indicates p < 0.05, and ** indicates p < 0.01.

[0066] The results showed that the antibacterial effect of Shouhui Tongbian composition on ATCC 700392 strain was as follows: at MIC, flagellar genes (flaA, flaB), adhesion genes (alpA, alpB, babA), virulence factors (cagA) and urease genes (ureE, ureF) were significantly downregulated after 24 hours of incubation. RT-qPCR analysis results showed that Shouhui Tongbian capsule had an effect on the mRNA level of virulence genes of Helicobacter pylori.

[0067] Example 6: Investigating the effect of the combined use of the Shouhui laxative composition and antibiotics

[0068] Determination method: Shouhui Tongbian Capsules (without capsule shells) were diluted, and the antibiotics were clarithromycin, metronidazole, amoxicillin, and levofloxacin. 60 μL of Helicobacter pylori ATCC 700392 bacterial suspension was added to each well of a 96-well plate, and at the same time, 30 μL of Shouhui Tongbian Capsules (0, 160, 320, 640, 1280, 2560 μg / mL) and 30 μL of antibiotics were added. The final bacterial concentration was approximately 1×106 CFU / mL. The concentration ranges of CLR, LEF, MTZ, and AMO were 0 - 0.008 μg / mL, 0 - 0.16 μg / mL, 0 - 6.4 μg / mL, and 0 - 0.4 μg / mL, respectively. It was placed in a triple-gas incubator and cultured with shaking at 37°C, in a microaerophilic environment, and at 150 rpm for 72 h. The concentration of the well that was clearly clarified to the naked eye was taken as the minimum combined inhibitory concentration. The combined inhibitory index FICI = MIC of Shouhui Tongbian Capsules (used alone) / MIC of Shouhui Tongbian Capsules (used in combination) + MIC of antibiotics (used alone) / MIC of antibiotics (used in combination). When FICI ≤ 0.5, it indicates a synergistic effect; when 0.5 < FICI ≤ 1, it is an additive effect; when 1 < FICI ≤ 2, it is an irrelevant effect; when FICI > 2, it is an antagonistic effect.

[0069] Table 3 Combined use of Shouhui Tongbian composition and antibiotics

[0070]

[0071] The results showed that there was no antagonistic effect in the combined application of the Shouhui Tongbian composition with the four antibiotics, clarithromycin, amoxicillin, levofloxacin, and metronidazole. It was an additive effect, which was beneficial to solving the problem of antibiotic resistance, reducing the dosage of antibiotics, and reducing the toxic and side effects.

Claims

1. Use of the Shouhui laxative composition in the preparation of an anti-Helicobacter pylori drug, characterized in that: The Shouhui laxative composition is composed of Polygonum multiflorum, Cassia seed, Citrus aurantium, aloe, donkey-hide gelatin, wolfberry, Atractylodes macrocephala and ginseng.

2. The use according to claim 1, characterized in that The use includes Helicobacter pylori infection or gastric ulcer and gastritis caused by Helicobacter pylori.

3. The use according to claim 1, characterized in that The use includes Helicobacter pylori type I and Helicobacter pylori type II infections.

4. The use according to claim 1, characterized in that The composition can be used in combination with antibiotics to prevent or treat Helicobacter pylori infection.

5. The use according to claim 4, characterized in that The antibiotic is selected from one or more of clarithromycin, amoxicillin, levofloxacin or metronidazole.

6. The use according to claim 1, characterized in that The composition can be used to prepare products including but not limited to medicines, health products and foods.

7. The use according to claim 1 or 6, characterized in that The composition may also contain pharmaceutically acceptable excipients and be prepared into a pharmaceutical preparation according to conventional preparation techniques.

8. The use according to claim 1 or 6, characterized in that The composition can be prepared as capsules, granules, pills or tablets.

9. The use according to claim 8, characterized in that The composition is Shouhui laxative capsule.

10. The use according to claim 1, characterized in that The minimum inhibitory concentration of the Shouhui laxative preparation is 320 μg / mL.

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