A kind of radix scutellariae extract for resisting Helicobacter pylori and its preparation method and compound medicine

Abstract

The invention discloses a radix scutellariae extract for resisting Helicobacter pylori, a preparation method thereof and a composite drug. The invention first obtains a radix scutellariae water extract by a water extraction method, and then extracts the radix scutellariae extract by using ethyl acetate as a solvent. In vitro experiments show that the method enriches the main active substances of radix scutellariae for resisting Helicobacter pylori, has a significant antibacterial effect on the standard strain SS1 of Helicobacter pylori, and has a minimum inhibitory concentration (MIC) of 32 μg / ml, which significantly inhibits the biofilm formation of the standard strain SS1 of Helicobacter pylori. In addition, it is also found that the radix scutellariae extract of the invention is used in combination with antibiotics commonly used in clinical treatment of HP infection, and has a synergistic effect with metronidazole and levofloxacin, and an additive effect with clarithromycin and amoxicillin, which has the effect of reducing the amount of antibiotics used in clinical application, and has potential application value for improving the current status of antibiotic resistance. Compared with the water extract, the MIC of the radix scutellariae extract extracted by the invention against Helicobacter pylori is greatly reduced, and the in vitro antibacterial activity is significantly enhanced.

Description

Technical Field

[0001] The invention belongs to the field of extracts and extraction technology, and particularly relates to a method for preparing a Helicobacter pylori-resistant Herba Lycopodii extract. Background Art

[0002] Helicobacter pylori (Helicobacterpylori, Hp) is a kind of spiral, flagellate, microaerophilic Gram-negative bacteria, it is located at various positions of stomach and duodenum, can continue to be colonized in human gastric mucosa, almost all can cause gastric mucosal active inflammation, a series of common diseases such as chronic gastritis, chronic atrophic gastritis, peptic ulcer and gastric cancer can also occur on the basis of active inflammation, long-term chronic inflammation can cause gastric mucosal morphology and pathological changes, if not taking therapeutic measures, cause intestinal metaplasia and dysplasia even canceration, seriously endanger public health. Helicobacter pylori has higher morbidity worldwide, is identified as class Ι carcinogen by the World Health Organization (WHO).

[0003] At present, the Hp eradication programs at home and abroad mainly include standard triple and quadruple therapy, and the antibacterial drugs mainly include metronidazole, clarithromycin, levofloxacin, etc. However, with the long-term use and abuse of antibiotics, Hp resistance is getting higher and higher, leading to intestinal flora imbalance, poor patient compliance, and large side effects. Traditional Chinese medicine has the characteristics of multi-component, multi-pathway, and multi-target action, and the incidence of drug resistance in traditional Chinese medicine therapy is low, adverse reactions are few, and patient compliance is good. Compared with traditional antibiotic therapy, traditional Chinese medicine therapy has unique advantages in anti-Hp. Therefore, it is of great significance to develop new anti-HP infection drugs from traditional Chinese medicine.

[0004] Xihuangcao is the dried whole herb of the plant of the genus Lamiaceae, also known as creek grass. It often grows on hillsides, fields, streams, etc. It tastes bitter and is cold in nature. It enters the liver, gallbladder, and large intestine meridians. It has the effects of clearing away heat and dampness, jaundice, cooling blood and dispersing blood stasis. It can be used to treat damp-heat jaundice, cholecystitis, trauma, sores, etc. The main active ingredients of Xihuangcao include diterpenes, flavonoids, phenolic acids, polysaccharides, coumarins, etc., which have multiple pharmacological activities such as liver protection, anti-cancer, antibacterial, anti-inflammatory, antiviral, antioxidant, and immunomodulatory. Abietanoids has long been used as a folk Chinese medicine to treat jaundice, hepatitis, acute cholecystitis and enteritis (Jun Wan, Hua-Yi Jiang, Jian-Wei Tang, et al. Ent-Abietanoids Isolated from Isodon serra[J]. Molecules, 2017, 22(02):309). It has a long history of use in southern regions such as Guangdong, Guangxi, Jiangxi, and Fujian. It is a commonly used Chinese herbal medicine in clinical and folk medicine, and is also the main raw material for a variety of health products and Chinese patent medicines, such as Compound Abietanoids Granules, Shiwei Abietanoids Granules, Danshitong Capsules, Xiaoyan Lidan Tablets, Shenling Gankang Capsules, etc. It is also an important ingredient in Cantonese herbal tea and has great market potential (Qiu Jiajia, Liu Junmin, Zhan Ruoting, Wang Deqin, Xu Youyang. Analysis of the main chemical components of different varieties of Abietanoids based on UPLC-Q-TOF-MS and UPLC-DAD[J]. Chinese Journal of Traditional Chinese Medicine, 2022, 47(13):3539-3547).

[0005] So far, no relevant research has been found on the anti-Helicobacter pylori infection effect of Xihuangcao extract. Summary of the invention

[0006] In view of the shortcomings of the existing technology for treating Helicobacter pylori, the present invention proposes a method for preparing a Herba Cynoglossi anti-Helicobacter pylori extract, which enriches the main active substances of Herba Cynoglossi anti-HP, thereby greatly improving the antibacterial activity.

[0007] To this end, the technical solution adopted in the present invention is:

[0008] The present invention provides a method for preparing an anti-Helicobacter pylori extract of Herba Lycopodii, comprising the following steps:

[0009] S1. Processing of medicinal materials: crush the Herba Lycopodii into coarse medicinal powder.

[0010] S2. Water extraction: extract the crude powder of the medicinal material in step S1 by water extraction to obtain the water extract of Herba Cynoglossi

[0011] S3, ethyl acetate extraction: using ethyl acetate as solvent, extracting the water extract of the herba scopariae in step S2 to obtain the herba scopariae extract.

[0012] Preferably, the ethyl acetate extraction method described in step S3 is: the ethyl acetate extraction method described in step S3 is: ethyl acetate is added to the water extract of the herb, and extraction is performed until there is no obvious color change between the aqueous layer and the organic layer, the extracts are combined, and concentrated by a rotary evaporator to obtain the herb extract.

[0013] Furthermore, the water extraction method of step S2 is: adding water to the crude powder of the medicinal material at a solid-liquid ratio of 1:10, then placing it under hot reflux extraction at 100° C. for 3 hours, collecting the filtrate and concentrating it to obtain the water extract of the Herba Lycopodii.

[0014] Preferably, in step S1, the crushed hyacinthus annuus is passed through a sieve No. 1 to No. 3 to obtain a coarse powder of the medicinal material. Specifically, the crushed hyacinthus annuus is passed through a sieve No. 2 to obtain a coarse powder of the medicinal material.

[0015] The present invention also provides an anti-Helicobacter pylori extract of Herba Cynoglossi from the preparation method; and application of the Herba Cynoglossi extract as a drug for preparing an anti-Helicobacter pylori drug.

[0016] Another technical solution of the present invention is to provide an anti-Helicobacter pylori compound medicine, comprising the anti-Helicobacter pylori Herba Lysimachiae extract prepared by the above solution, and at least one of metronidazole, levofloxacin, clarithromycin and amoxicillin.

[0017] Furthermore, the above-mentioned anti-Helicobacter pylori compound medicine is characterized in that the anti-Helicobacter pylori Xihuangcao extract is composed of the anti-Helicobacter pylori Xihuangcao extract and metronidazole in a mass ratio of 1:0.0125; or is composed of the anti-Helicobacter pylori Xihuangcao extract and clarithromycin in a mass ratio of 1:0.0039; or is composed of the anti-Helicobacter pylori Xihuangcao extract and amoxicillin in a mass ratio of 1:0.00049; or is composed of the anti-Helicobacter pylori Xihuangcao extract and levofloxacin in a mass ratio of 1:0.0156.

[0018] Compared with the prior art, the use of the Herba Cynoglossi extract in the preparation of anti-Helicobacter pylori drugs disclosed in the present invention is disclosed for the first time. The Herba Cynoglossi extract has inhibitory activity against Helicobacter pylori and has an inhibitory effect on the formation of Helicobacter pylori biofilm, and can be used for the prevention and treatment of Helicobacter pylori infection.

[0019] In vitro experiments show that the extract has strong anti-Helicobacter pylori activity, has a significant antibacterial effect on the standard strain of Helicobacter pylori SS1, and the minimum inhibitory concentration (MIC) is 32μg / ml, and the inhibitory effect on HP is concentration-dependent, and the greater the concentration, the more obvious the inhibitory effect. The extract can also significantly inhibit the biofilm formation of the standard strain of Helicobacter pylori SS1. In addition, it is found that the Xihuangcao extract of the present invention is used in combination with antibiotics commonly used in the clinical treatment of HP infection, and has a synergistic effect with metronidazole and levofloxacin, and an additive effect with clarithromycin and amoxicillin, which has the effect of reducing the amount of clinically used antibiotics, and has potential application value for improving the current status of antibiotic resistance. The minimum inhibitory concentration (MIC) of the water extract is 1024μg / ml. In contrast, the MIC of the Xihuangcao extract extracted by the present invention against Helicobacter pylori is greatly reduced, and the in vitro antibacterial activity is significantly enhanced. The Xihuangcao extract obtained by the method of the present invention can enrich the main active substances of Xihuangcao against Hp, thereby improving the antibacterial activity of the Xihuangcao extract. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The effect of the Xihuangcao extract of the present invention on the growth of HP SS1 after 72 hours of action;

[0021] Figure 2 is the time-inhibition curve of the Herba Cynoglossi extract of the present invention against HP SS1;

[0022] Figure 3 This is a crystal violet staining diagram of the Herba Lycopodii extract of the present invention:

[0023] Figure 4 The present invention relates to the effect of the Herba Lysimachiae extract on the biofilm formation of HP SS1. DETAILED DESCRIPTION

[0024] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in each embodiment of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0025] Unless otherwise specified, the equipment used in this example is conventional experimental equipment, the materials and reagents used are commercially available unless otherwise specified, and the experimental methods without special instructions are also conventional experimental methods.

[0026] Herba Cynanchifoliae was purchased from Anhui Huifeng Pharmaceutical Co., Ltd.

[0027] Example 1

[0028] The present embodiment provides a method for preparing a Helicobacter pylori-resistant Herba Lycopodii extract, comprising the following steps:

[0029] S1. Clean the Xihuangcao powder, dry it in the sun, crush it into coarse powder, and pass it through a No. 1 sieve to obtain coarse powder;

[0030] S2. Water extraction: add water to the crude powder of the medicinal material at a solid-liquid ratio of 1:14, then reflux and extract at 100°C for 3 hours, collect the filtrate to obtain the water extract of Rhizoma Cynoglossi

[0031] S3, ethyl acetate extraction: add ethyl acetate twice the volume of the water extract of S2 tricholoma scoparia, extract until there is no obvious color change between the aqueous layer and the organic layer, combine the extracts, and concentrate on a rotary evaporator to obtain the tricholoma scoparia extract.

[0032] Example 2

[0033] The present embodiment provides a method for preparing a Helicobacter pylori-resistant Herba Lycopodii extract, comprising the following steps:

[0034] S1. Clean the Xihuangcao powder, dry it in the sun, crush it into coarse powder, and pass it through a No. 2 sieve to obtain coarse powder;

[0035] S2. Water extraction: add water to the crude powder of the medicinal material at a solid-liquid ratio of 1:13, then reflux and extract at 90°C for 4 hours, collect the filtrate to obtain the water extract of the Herba Lycopodii.

[0036] S3, ethyl acetate extraction: add ethyl acetate in an amount 3 times the volume of the water extract of S2 tricholoma, extract until there is no obvious color change between the aqueous layer and the organic layer, combine the extracts, and concentrate on a rotary evaporator to obtain the tricholoma extract.

[0037] Example 3

[0038] The present embodiment provides a method for preparing a Helicobacter pylori-resistant Herba Lycopodii extract, comprising the following steps:

[0039] S1. Clean the herb, dry it in the sun, and grind it into powder using a grinder. Sieve it through a No. 3 sieve to obtain a coarse powder of the medicinal material.

[0040] S2. Water extraction: add water to the crude powder of the medicinal material at a solid-liquid ratio of 1:12, then reflux and extract at 110°C for 1 hour, collect the filtrate to obtain the water extract of the Herba Lycopodii.

[0041] S3, ethyl acetate extraction: add ethyl acetate in an amount 3 times the volume of the water extract of S2 tricholoma, extract until there is no obvious color change between the aqueous layer and the organic layer, combine the extracts, and concentrate on a rotary evaporator to obtain the tricholoma extract.

[0042] Example 4

[0043] The present embodiment provides a method for preparing a Helicobacter pylori-resistant Herba Lycopodii extract, comprising the following steps:

[0044] S1. Clean the herb, dry it in the sun, and grind it with a grinder. Sieve it through a No. 2 sieve to obtain a coarse powder of the medicinal material.

[0045] S2. Water extraction: add water to the crude powder of the medicinal material at a solid-liquid ratio of 1:10, then reflux and extract at 100°C for 3 hours, collect the filtrate to obtain the water extract of the Herba Lycopodii.

[0046] S3, ethyl acetate extraction: add ethyl acetate in an amount 3 times the volume of the water extract of S2 tricholoma, extract until there is no obvious color change between the aqueous layer and the organic layer, combine the extracts, and concentrate on a rotary evaporator to obtain the tricholoma extract.

[0047] Comparative Example 1

[0048] The preparation method of a Herba Lycopodiellae extract for resisting Helicobacter pylori provided in this comparative example comprises the following steps:

[0049] S1. Clean the herb, dry it in the sun, and grind it with a grinder. Sieve it through a No. 2 sieve to obtain a coarse powder of the medicinal material.

[0050] S2. Water extraction: add water to the crude powder of the medicinal material at a solid-liquid ratio of 1:10, then reflux and extract at 100°C for 3 hours, collect the filtrate to obtain the water extract of the Herba Lycopodii.

[0051] Comparative Example 2

[0052] The preparation method of a Herba Lycopodiellae extract for resisting Helicobacter pylori provided in this comparative example comprises the following steps:

[0053] S1. Clean the herb, dry it in the sun, and grind it with a grinder. Sieve it through a No. 2 sieve to obtain a coarse powder of the medicinal material.

[0054] S2. Alcohol extraction: add 90% ethanol to the crude powder of the medicinal material at a solid-liquid ratio of 1:10, then reflux and extract at 100°C for 3 hours, collect the filtrate to obtain the alcohol extract of the Herba Lycopodiellae.

[0055] Comparative Example 3

[0056] The preparation method of a Herba Lycopodiellae extract for resisting Helicobacter pylori provided in this comparative example comprises the following steps:

[0057] S1. Clean the herb, dry it in the sun, and grind it with a grinder. Sieve it through a No. 2 sieve to obtain a coarse powder of the medicinal material.

[0058] S2. Add 90% ethanol to the crude powder of the medicinal material at a solid-liquid ratio of 1:10, then place it under hot reflux extraction at 100°C for 2 hours, and collect the filtrate to obtain the alcohol extract of the Herba Cynoglossi.

[0059] S3, ethyl acetate extraction: add ethyl acetate in an amount 3 times the volume of the ethanol extract of angelica dahurica to the ethanol extract of angelica dahurica in S2, extract until there is no obvious color change between the aqueous layer and the organic layer, combine the extracts, and concentrate them on a rotary evaporator to obtain the angelica dahurica extract.

[0060] In order to verify the effect of the technical solution provided in the present application, the in vitro activity against Helicobacter pylori in Example 4 and Comparative Examples 1-3 was tested below.

[0061] 1.1 Materials

[0062] 1) Strain: Helicobacter pylori standard strain SS1. HP SS1 was donated by Professor Chen Ye of Southern Medical University and was stored in a -80°C refrigerator.

[0063] 2) Culture medium and main reagents: brain heart infusion (BHI), Columbia medium, fetal calf serum (FCS) and anhydrous ethanol.

[0064] 3) Main instruments: constant temperature incubator, multifunctional microplate reader, centrifuge and electronic balance, etc.

[0065] 1.2 Determination method: Prepare multiple concentration gradients of Herba Cynoglossi extract solutions (512 μg / mL, 256 μg / mL, 128 μg / mL, 64 μg / mL, 32 μg / mL, etc.) by the two-fold dilution method, and prepare them with BHI culture medium containing 10% FCS. Pipette 50 μL of the above solution and add it to a 96-well plate. Set up 3 replicate wells for each drug concentration, and set up sterile control wells (containing 10% FCS BHI culture medium, no strain inoculation) and growth control wells (not containing any drugs, only inoculating strains). Take out the strain from the incubator, scrape the bacterial moss with a sterile loop to make a bacterial suspension in a BHI culture solution containing 10% FCS, adjust the turbidity of the bacterial suspension to 1McF with BHI culture solution, dilute 10 times, and inoculate 50μL per well in the above-mentioned 96-well plate containing the drug, immediately place the 96-well plate in an anaerobic box, add a microaerobic bag, and culture at 37°C for 72h, observe the results and test the OD600 absorbance value. When the bacteria in the growth control well (i.e., without drug) grow significantly and the sterile control group grows aseptically, and the trend of the OD600 absorbance value test result is consistent with the turbidity observed by naked eyes, the experiment is meaningful, and the minimum drug concentration that can be observed in the clear well is the MIC, and the experiment is repeated 3 times.

[0066] The results are shown in Table 1. As can be seen from Table 1, the MIC of Example 4 is lower than that of Comparative Examples 1-3, indicating that the extract obtained by this method is enriched with the active ingredients of Herba Cynoglossi anti-HP, thereby enhancing its antibacterial activity. Figure 1It can be seen that the sterile control wells are clear and the growth control wells are obviously turbid. This experimental result is meaningful. Through visual observation, the MIC of Example 4 is 32 μg / mL.

[0067] Table 1 Minimum inhibitory concentration (MIC) of extracts obtained by different extraction methods against Hp SS1

[0068]

[0069] Example 5

[0070] An anti-Helicobacter pylori extract of Herba Cynoglossi was prepared from the anti-Helicobacter pylori extract of Herba Cynoglossi in Example 4 and metronidazole in a mass ratio of 1:0.0125;

[0071] Example 6

[0072] An anti-Helicobacter pylori extract of Herba Cynoglossi was prepared from the anti-Helicobacter pylori extract of Herba Cynoglossi in Example 4 and clarithromycin in a mass ratio of 1:0.0039;

[0073] Example 7

[0074] An anti-Helicobacter pylori radix cyperi extract is composed of the anti-Helicobacter pylori radix cyperi extract prepared in Example 4 and amoxicillin in a mass ratio of 1:0.00049;

[0075] Example 8

[0076] An anti-Helicobacter pylori Herba Lysimachiae extract is composed of the anti-Helicobacter pylori Herba Lysimachiae extract prepared in Example 4 and levofloxacin in a mass ratio of 1:0.0156.

[0077] The effect of Herba Lycopodiellae extract combined with antibiotics on Hp SS1 was determined by the checkerboard dilution method.

[0078] 1. Test samples: the extract of the present invention, amoxicillin, clarithromycin, metronidazole, levofloxacin

[0079] 2. Determination method: Prepare the Herba Rabdosiae Rubescentis extract solutions with multiple concentration gradients (128 μg / mL, 64 μg / mL, 32 μg / mL, 16 μg / mL) according to the two-fold dilution method, and prepare antibiotic solutions with different concentrations in the same way (amoxicillin 0.031 - 0.25 μg / mL, clarithromycin 0.002 - 0.015 μg / mL, metronidazole 1 - 8 μg / mL, levofloxacin 0.25 - 2 μg / mL). Take out the strain SS1 from the incubator, scrape the bacteria into the broth with a sterilized loop, adjust the turbidity of the bacterial suspension to 1 McF with BHI culture medium containing 10% FCS and then dilute it 10 times. According to the microbroth checkerboard dilution method, inoculate 100 μL of the mixed solution of bacterial suspension - Herba Rabdosiae Rubescentis extract solution - antibiotic solution into each well of a 96-well plate at a ratio of bacterial suspension: Herba Rabdosiae Rubescentis extract solution: antibiotic solution = 2:1:1. Then place it in an anaerobic box and add a microaerophilic bag, and culture at 37 °C for 72 h. The concentration at which obvious clear wells can be observed is the minimum combined inhibitory concentration. The combined inhibitory index FICI = MIC (MIC of Herba Rabdosiae Rubescentis in combination) / MIC (MIC of Herba Rabdosiae Rubescentis administered alone) + MIC (MIC of the antibiotic in combination) / MIC (MIC of the antibiotic administered alone). When FICI ≤ 0.5, it is a synergistic effect; when 0.5 < FICI ≤ 1, it is an additive effect; when 1 < FICI ≤ 4, it is an indifferent effect, and when FICI > 4.0, it is an antagonistic effect.

[0080] As can be seen from Table 2, when the Herba Rabdosiae Rubescentis extract of the present invention is used in combination with the antibiotics commonly used in clinical treatment of Hp infection, it shows a synergistic effect with metronidazole and levofloxacin, and an additive effect with clarithromycin and amoxicillin. The combination can reduce the MIC of the antibiotics, has the effect of reducing the dosage of antibiotics used clinically, and has potential application value for improving the current situation of antibiotic resistance.

[0081] Table 2 Effects of the combination of Herba Rabdosiae Rubescentis extract and antibiotics against Hp

[0082]

[0083] Dynamically monitor the antibacterial effect of Herba Rabdosiae Rubescentis extract on HP SS1 through the time-bacteriostatic curve

[0084] 1. Test sample: The Herba Rabdosiae Rubescentis extract of the present invention.

[0085] 2. Determination method: Prepare multiple concentration gradients of the Herba Lycopodiellae extract solution (256 μg / mL, 128 μg / mL, 64 μg / mL, 32 μg / mL, 16 μg / mL) by the two-fold dilution method, and use the broth without the solution (drug-free broth) as the growth control group. Take out strain SS1 from the incubator, scrape the bacteria with a sterile loop and put them into brain heart infusion to make a bacterial suspension, adjust the turbidity of the bacterial suspension to 1McF with BHI culture medium containing 10% FCS, and then dilute it 10 times, inoculate 2mL of the mixture of bacterial suspension-drug solution / drug-free broth in each well of a 6-well plate at the ratio of bacterial suspension: drug solution / drug-free broth = 1:1, then place it in an anaerobic box, add a microaerobic bag, and culture at 37°C, aspirate 100μL of culture solution to measure OD600nm at 0h, 12h, 24h, 36h, 48h, 60h, and 72h. The larger the absorbance value, the more bacteria there are.

[0086] 3. By Figure 2 It can be seen that the inhibitory effect of the Herba Lycopodii extract on Hp is concentration-dependent. The higher the concentration of the Herba Lycopodii extract, the more obvious the inhibitory effect. When the concentration of the Herba Lycopodii extract is 32 μg / mL, the growth of Hp is basically inhibited.

[0087] Effects of extracts from Rhizoma Cyperi on biofilm formation of HP SS1

[0088] 1. Test sample: the Herba Lysimachiae extract of the present invention.

[0089] 2. Determination method:

[0090] 200 μL of bacterial suspension with a concentration of 1×106 CFU / mL was added to each group of the 48-well plate, and cultured for 72 hours at 37°C microaerobic and saturated humidity to form Hp biofilm. The culture supernatant was removed, and then fresh culture medium was added to wash and remove suspended Hp. Multiple concentration gradients of Xihuangcao extract solutions (256 μg / mL, 128 μg / mL, 64 μg / mL, 32 μg / mL, 16 μg / mL) were prepared by the two-fold dilution method, and BHI without the solution was used as the growth control group, and incubated for 24 hours at 37°C microaerobic conditions. At the end of the incubation, the culture supernatant was removed, and the culture plate was washed with 200 μL of PBS to remove bacterial non-adherent cells. After drying, it was stained with 0.1% (w / v) crystal violet at room temperature for 15 minutes. The crystal violet staining solution was discarded, and the samples were washed with PBS until no color remained. After drying at room temperature, 75% ethanol solution was added to dissolve the samples, and the absorbance at 570 nm was measured using an enzyme marker to measure the amount of biofilm.

[0091] 3. By Figure 4It can be seen that 256μg / mL, 128μg / mL, and 64μg / mL of Herba Lysimachiae extracts can significantly inhibit the formation of Hp biofilm (p**<0.01, p****<0.001).

[0092] The embodiments of the present invention are described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions and variations of these embodiments are made without departing from the principles and spirit of the present invention, and still fall within the protection scope of the present invention.

Claims

1. A method for preparing an anti-Helicobacter pylori extract of Herba Lycopodii, characterized in that: The following steps are involved: S1. Grind the herb into coarse powder; S2, water extraction: extracting the crude powder of the medicinal material in step S1 by water extraction to obtain a water extract of the herb; S3, ethyl acetate extraction: using ethyl acetate as solvent, extracting the water extract of the herba scopariae in step S2 to obtain the herba scopariae extract.

2. The method for preparing an anti-Helicobacter pylori extract of Herba Lycopodii according to claim 1, characterized in that: The ethyl acetate extraction method described in step S3 is: adding ethyl acetate to the water extract of the herb, extracting until there is no obvious color change between the water layer and the organic layer, combining the extracts, and concentrating them on a rotary evaporator to obtain the herb extract.

3. The method for preparing an anti-Helicobacter pylori extract of Herba Lycopodii according to claim 2, characterized in that: The volume ratio of the ethyl acetate to the water extract of the herb is (2-4):

1.

4. The method for preparing an anti-Helicobacter pylori extract of Herba Cynoglossifoliae according to claim 1, characterized in that: The water extraction method of step S2 is: adding water to the crude powder of the medicinal material at a solid-liquid ratio of 1: (10-14), and then placing it at (90-110° C.) for reflux extraction for (1-4) hours, and collecting the filtrate to obtain the water extract of the herb.

5. The method for preparing the anti-Helicobacter pylori extract of Herba Lycopodii according to claim 4, characterized in that: The water extraction method of step S2 is: adding water to the crude powder of the medicinal material at a solid-liquid ratio of 1:10, then placing it under hot reflux extraction at 100° C. for 3 hours, collecting the filtrate and concentrating it to obtain the water extract of the Herba Lycopodii.

6. The method for preparing the anti-Helicobacter pylori extract of Herba Lycopodii according to claim 5, characterized in that: The concentration is to concentrate until the solid-liquid ratio of the crude medicinal material powder to the filtrate is 1:

3.

7. The method for preparing the anti-Helicobacter pylori extract of Herba Lycopodii according to claim 1, characterized in that: Step S1, the crushed Herba Cynanchum wilfordii is sieved through No. 1 to No. 3 sieves to obtain a coarse powder of the medicinal material.

8. The anti-Helicobacter pylori extract of Herba Lycopodii obtained by the preparation method according to any one of claims 1 to 7, characterized in that: The extract of the Herba Cynoglossi is used as a drug for preparing drugs against Helicobacter pylori.

9. An anti-Helicobacter pylori compound medicine comprising the anti-Helicobacter pylori Herba Cynoglossi extract of claim 8, and at least one of metronidazole, levofloxacin, clarithromycin, and amoxicillin.

10. The anti-Helicobacter pylori compound medicine according to claim 9, characterized in that: The anti-Helicobacter pylori Xihuangcao extract is composed of the anti-Helicobacter pylori Xihuangcao extract of claim 8 and metronidazole in a mass ratio of 1:0.0125; or the anti-Helicobacter pylori Xihuangcao extract of claim 8 and clarithromycin in a mass ratio of 1:0.0039; or the anti-Helicobacter pylori Xihuangcao extract of claim 8 and amoxicillin in a mass ratio of 1:0.00049; or the anti-Helicobacter pylori Xihuangcao extract of claim 8 and levofloxacin in a mass ratio of 1:0.0156.

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