A spleen-strengthening and stomach-benefiting prescription for treating gastric precancerous lesions, and its preparation method and application

Abstract

The present invention mainly relates to a spleen-tonifying and stomach-benefiting prescription for treating gastric precancerous lesions, as well as its preparation method and application, and belongs to the field of biomedicine technology. The spleen-tonifying and stomach-benefiting prescription described in the present invention is a traditional Chinese medicine composition, which includes ginseng, stir-fried white atractylodes, astragalus, Panax notoginseng, dried ginger, tangerine peel, ginger pinellia, poria, white flower oldenlandia diffusa, vinegar-curdled zedoaria, ganoderma lucidum, and licorice. The above raw materials are extracted with water to obtain the traditional Chinese medicine composition of the present invention. The traditional Chinese medicine composition of the present invention can alleviate the bloating symptoms and related pathological changes of gastric precancerous lesions; inhibit the overexpression of DMBT1, WFDC2, CFTR, AQP5, and CLU, which are markers of malignant progression of gastric precancerous lesions; reduce atrophy and intestinal metaplasia of the gastric antrum and gastric angle, effectively treat gastric precancerous lesions, and prevent the occurrence of gastric cancer.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to a spleen-strengthening and stomach-tonifying prescription for treating gastric precancerous lesions, and a preparation method and application thereof. Background Art

[0002] Gastric cancer is one of the common malignant tumors, which seriously endangers human health. Gastric precancerous lesions are an important pathological concept in the progression of gastric cancer, which refers to certain pathological changes in the gastric mucosa, which are more likely to turn into gastric cancer. Gastric mucosal cancer does not directly transform from normal cells into cancer cells, but needs to go through a multi-step process of canceration, namely chronic superficial gastritis → atrophic gastritis → intestinal metaplasia → dysplasia → gastric cancer. The lesions that appear during this period are called precancerous lesions. Its etiology and pathogenesis have not yet been fully elucidated. It is now clear that Helicobacter pylori infection is the main cause of chronic gastritis, but other physical, chemical and biological harmful factors can also cause this disease if they act on susceptible humans for a long time. Common clinical manifestations include stomach bloating, stomach pain, indigestion and other symptoms. Gastric bleeding may also occur. Sometimes, due to the destruction of intrinsic gastric factors, vitamin B 12 Malabsorption can lead to anemia. Currently, gastric precancerous lesions are primarily treated with medication, which requires regular follow-up. However, medications often provide limited control, necessitating surgery, making complete cure difficult and potentially long-term. Therefore, there is an urgent need for a drug that can effectively treat gastric precancerous lesions and improve their treatment and control. Summary of the Invention

[0003] In view of this, the object of the present invention is to provide a traditional Chinese medicine composition for treating gastric precancerous lesions. The traditional Chinese medicine composition is named Jianpi Yiwei Fang, which can improve the bloating symptoms of gastric precancerous lesions, alleviate the pathological changes of gastric precancerous lesions, inhibit the overexpression of malignant progression markers of gastric precancerous lesions, and avoid the occurrence of gastric cancer.

[0004] The invention provides a traditional Chinese medicine composition for treating gastric precancerous lesions. The composition comprises the following raw materials in parts by weight: 85-95 parts of ginseng, 115-125 parts of stir-fried atractylodes macrocephala, 145-155 parts of astragalus, 25-35 parts of notoginseng, 95-105 parts of dried ginger, 95-105 parts of tangerine peel, 85-95 parts of ginger pinellia, 145-155 parts of poria, 95-105 parts of oldenlandia diffusa, 85-95 parts of vinegared zedoaria, 115-125 parts of ganoderma lucidum and 95-105 parts of liquorice.

[0005] Preferably, the Chinese medicine composition comprises the following raw materials in parts by weight: 88-92 parts of ginseng, 118-122 parts of stir-fried Atractylodes macrocephala, 148-152 parts of Astragalus membranaceus, 28-32 parts of Panax notoginseng, 98-102 parts of dried ginger, 98-102 parts of dried tangerine peel, 88-92 parts of ginger and Pinellia ternata, 148-152 parts of Poria cocos, 98-102 parts of Hedyotis diffusa, 88-92 parts of vinegared Curcuma zedoaria, 118-122 parts of Ganoderma lucidum and 98-102 parts of licorice.

[0006] The present invention provides a method for preparing the traditional Chinese medicine composition, comprising the following steps:

[0007] Ginseng, stir-fried Atractylodes macrocephala, Astragalus membranaceus, dried ginger, tangerine peel, Pinellia ternata, Poria cocos, Hedyotis diffusa, Curcuma zedoaria, Ganoderma lucidum and licorice are mixed to obtain a mixture, the mixture is mixed with water, soaked, and decocted to obtain extract 1;

[0008] Grind Panax notoginseng, mix with water, soak, and boil to obtain extract 2;

[0009] The extract 1 and the extract 2 are mixed to obtain the traditional Chinese medicine composition.

[0010] Preferably, the mass volume ratio of the mixture to water is 1 g: (8-12) mL; the mass volume ratio of Panax notoginseng to water is 1 g: (15-18) mL.

[0011] Preferably, the soaking time is 55 to 65 minutes.

[0012] Preferably, when extract 1 is obtained, the number of decoction extractions is ≥ 2 times.

[0013] Preferably, the decoction extraction is: heating to boiling and then decocting for 35 to 45 minutes.

[0014] The present invention also provides application of the traditional Chinese medicine composition or the preparation method in preparing medicine for treating gastric precancerous lesions.

[0015] Preferably, the gastric precancerous lesion comprises spasmolytic polypeptide-expressing metaplasia.

[0016] Preferably, the gastric precancerous lesions include chronic atrophic gastritis.

[0017] Beneficial effects of the present invention:

[0018] The invention provides a spleen-strengthening and stomach-benefiting prescription for treating gastric precancerous lesions. The spleen-strengthening and stomach-benefiting prescription is a traditional Chinese medicine composition. In the traditional Chinese medicine composition, ginseng greatly replenishes vital energy, strengthens the spleen and nourishes the stomach, astragalus is sweet and warm, replenishes qi and strengthens the spleen, and stir-fried white atractylodes is sweet, bitter and warm, strengthens the spleen and dries dampness to help transportation and transformation. The three are collectively monarch medicines; tangerine peel and ginger pinellia regulate qi and harmonize the stomach, relieve fullness and eliminate abdominal distension, panax notoginseng and vinegar-curcuma activate blood circulation, remove blood stasis, eliminate accumulation and relieve pain, and white flower hedyotis diffusa herb clears heat and detoxifies, and are collectively ministerial medicines; dried ginger warms the middle and dispels cold, poria strengthens the spleen and eliminates dampness and tranquilizes the mind, and ganoderma lucidum replenishes qi and nourishes the mind, and are collectively adjuvant medicines; licorice strengthens the spleen and harmonizes the middle and harmonizes the various medicines, and is collectively a guiding medicine; the whole prescription has both tonifying and purging effects, uses both cold and warm effects, and treats both the symptoms and the root cause, with strict compatibility, and has the effects of strengthening the spleen and replenishing qi, and promoting blood circulation and benefiting the stomach. It is used to treat gastric precancerous lesions, can significantly alleviate the bloating symptoms and related pathological changes of gastric precancerous lesions, restore chief cell-like characteristics, and significantly reduce the level of inflammatory cell infiltration; inhibit the overexpression of DMBT1, WFDC2, CFTR, AQP5 and CLU, markers of malignant progression of gastric precancerous lesions; reduce atrophy and intestinal metaplasia of the gastric antrum and gastric angle, effectively treat gastric precancerous lesions, and avoid the occurrence of gastric cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 is the extracted ion chromatogram of the components to be tested; wherein A is the reference substance, and B is the freeze-dried powder prepared in Example 1; the blue numbers represent: 1 is deacylated arginine, 2 is liquiritin, 3 is hesperidin, 4 is notoginsenoside R1, 5 is ginsenoside Re, 6 is ginsenoside Rg1, 7 is ginsenoside Rb1, 8 is ganoderic acid A, 9 is glycyrrhizic acid, 10 is astragaloside IV, 11 is 6-gingerol, 12 is atractylodes lactone II, and 13 is atractylodes lactone I.

[0020] Figure 2 The figures show the pathological effects of the Chinese medicine composition (Jianpi Yiwei Fang) of the present invention on preventing and treating tamoxifen-induced SPEM gastric precancerous lesions in mice; wherein, C is the control group, M is the model group, Y is the positive drug group, GL is the Jianpi Yiwei Fang low-dose group, GM is the Jianpi Yiwei Fang medium-dose group, and GH is the Jianpi Yiwei Fang high-dose group.

[0021] Figure 3 Figure 2 is the effect of the Chinese medicine composition of the present invention on the expression of genes related to the malignant progression of SPEM gastric precancerous lesions; where # indicates P < 0.05 compared with C, ## indicates P < 0.01 compared with C, * indicates P < 0.05 compared with M, and ** indicates P < 0.01 compared with M; n = 8, data are based on express.

[0022] Figure 4 These are the results of gastroscopy and pathology examination of Case 1 in March 2024; from top to bottom, they are the results of gastric antrum gastroscopy examination and the results of gastric antrum pathology examination.

[0023] Figure 5 These are the results of gastroscopy and pathology examination of Case 1 in November 2024; from top to bottom, they are the results of gastric antrum gastroscopy examination and the results of gastric antrum pathology examination.

[0024] Figure 6 These are the results of gastroscopy and pathological examination of Case 2 in July 2024; from top to bottom they are the results of gastroscopy examination of the gastric angle, the results of pathological examination of the gastric angle, the results of gastroscopy examination of the gastric antrum, and the results of pathological examination of the gastric antrum.

[0025] Figure 7 These are the results of gastroscopy and pathological examination of Case 2 in December 2024; from top to bottom they are the results of gastroscopy of the gastric angle, the results of pathological examination of the gastric angle, the results of gastroscopy of the gastric antrum, and the results of pathological examination of the gastric antrum.

[0026] Figure 8 These are the results of gastroscopy and pathological examination of Case 3 in June 2023; from top to bottom they are the results of gastroscopy examination of the gastric angle, the results of pathological examination of the gastric angle, the results of gastroscopy examination of the gastric antrum, and the results of pathological examination of the gastric antrum.

[0027] Figure 9 These are the results of gastroscopy and pathology examination of Case 3 in August 2024; from top to bottom, they are the gastroscopy results of the gastric angle (left) and gastric antrum (right), and the pathology examination results of the gastric antrum. DETAILED DESCRIPTION

[0028] The present invention provides a traditional Chinese medicine composition for treating gastric precancerous lesions, comprising the following raw materials in parts by weight: 85-95 parts of ginseng, 115-125 parts of stir-fried atractylodes macrocephala, 145-155 parts of astragalus, 25-35 parts of notoginseng, 95-105 parts of dried ginger, 95-105 parts of dried tangerine peel, 85-95 parts of ginger pinellia, 145-155 parts of poria, 95-105 parts of oldenlandia diffusa, 85-95 parts of vinegared zedoaria, 115-125 parts of ganoderma lucidum and 95-105 parts of liquorice; preferably comprising the following raw materials in parts by weight: 88-92 parts of ginseng, 118-122 parts of stir-fried atractylodes macrocephala, 148-155 parts of astragalus. 152 parts, 28-32 parts of Panax notoginseng, 98-102 parts of dried ginger, 98-102 parts of dried tangerine peel, 88-92 parts of ginger pinellia, 148-152 parts of Poria, 98-102 parts of white flower oldenlandia diffusa, 88-92 parts of vinegared zedoary, 118-122 parts of ganoderma lucidum and 98-102 parts of liquorice; more preferably, the following raw materials in parts by weight are included: 90 parts of ginseng, 120 parts of stir-fried atractylodes, 150 parts of astragalus, 30 parts of Panax notoginseng, 100 parts of dried ginger, 100 parts of dried tangerine peel, 90 parts of ginger pinellia, 150 parts of Poria, 100 parts of white flower oldenlandia diffusa, 90 parts of vinegared zedoary, 120 parts of ganoderma lucidum and 100 parts of liquorice.

[0029] The present invention has no special limitation on the sources of the ginseng, stir-fried atractylodes macrocephala, astragalus root, notoginseng, dried ginger, tangerine peel, ginger pinellia, poria, oldenlandia diffusa, vinegared zedoaria, ganoderma lucidum and liquorice, and conventional commercial products in the field can be used.

[0030] In the present invention, ginseng greatly replenishes vital energy, strengthens the spleen and nourishes the stomach; astragalus is sweet and warm, replenishes qi and nourishes the spleen; stir-fried white atractylodes is sweet, bitter and warm in nature, strengthens the spleen and dries dampness to help transportation and transformation; tangerine peel, ginger and pinellia regulate qi and harmonize the stomach, relieve fullness and eliminate abdominal distension; Panax notoginseng and vinegar-curcuma promote blood circulation, remove blood stasis, eliminate accumulation and relieve pain; white flower hedyotis diffusa herb clears away heat and detoxifies; dried ginger warms the middle and dispels cold; poria strengthens the spleen and eliminates dampness, and calms the mind and soothes the nerves; ganoderma lucidum replenishes qi and nourishes the spirit; licorice strengthens the spleen and harmonizes the middle, and harmonizes all the medicines.

[0031] This invention uses ginseng, astragalus, and stir-fried atractylodes as main ingredients; tangerine peel, ginger pinellia, Panax notoginseng, vinegared zedoaria, and Hedyotis diffusa as assistant ingredients; dried ginger, poria, and ganoderma lucidum as adjuvant ingredients; and liquorice as a guiding ingredient. The formula has both tonifying and purging properties, treating both the symptoms and the root cause, with a rigorous combination of ingredients. It has the effects of strengthening the spleen and replenishing qi, promoting blood circulation, and benefiting the stomach. It is effective in treating gastric precancerous lesions, particularly chronic atrophic gastritis.

[0032] The present invention also provides a method for preparing the Chinese medicine composition, comprising the following steps:

[0033] First, prepare a mixture of ginseng, stir-fried atractylodes macrocephala, astragalus root, dried ginger, tangerine peel, ginger pinellia, poria, hedyotis diffusa, vinegared zedoaria, ganoderma lucidum and licorice, mix the mixture with water, soak it, and boil it to obtain extract 1;

[0034] In the present invention, the mass-to-volume ratio of the mixture to water is preferably 1g:(8-12)mL. In one embodiment, it can be 1g:8mL, 1g:9mL, 1g:10mL, 1g:11mL, or 1g:12mL. After the mixture is mixed with water, it is soaked for a period of preferably 55-65 minutes. In one embodiment, it can be soaked for 55 minutes, 58 minutes, 60 minutes, 63 minutes, or 65 minutes. After soaking, the mixture is decocted for extraction. The number of decoction extractions is preferably ≥2 times. In one embodiment, it can be decocted for 2, 3, or 4 times. The decoction extraction is preferably performed by heating to boiling and then decocting for 35-45 minutes. In one embodiment, it can be heated to boiling and then decocted for 35 minutes, 38 minutes, 40 minutes, 42 minutes, or 45 minutes. The decoction is preferably performed at a slight boil. In the present invention, after each decoction extraction, the decoction liquid is poured out, and water is added to the medicinal residue for the next decoction. After the decoction is completed, the decoction liquids of each decoction are combined, filtered, and the filtrate is collected to obtain Extract 1. The present invention does not specifically limit the filtering method, and conventional filtering methods in the art can be used.

[0035] Grind Panax notoginseng, mix with water, soak, and boil to obtain extract 2;

[0036] In the present invention, the Panax notoginseng is preferably pulverized into a powder, preferably an ultrafine powder, which means that the powder can completely pass through a No. 8 sieve and contains at least 95% of the powder that can pass through a No. 9 sieve. The present invention does not specifically limit the method of pulverization; conventional pulverization methods in the art can be used. In one embodiment, grinding can be used. The pulverized Panax notoginseng is mixed and soaked in water. The mass-to-volume ratio of Panax notoginseng to water is preferably 1g:(15-18)mL. In one embodiment, the ratio can be 1g:15mL, 1g:15.5mL, 1g:16mL, 1g:16.7mL, 1g:17mL, or 1g:18mL. The soaking time is preferably 55-65 minutes. In one embodiment, the soaking time can be 55 minutes, 58 minutes, 60 minutes, 63 minutes, or 65 minutes. After soaking, the Panax notoginseng is decocted for extraction. The decocting extraction is preferably performed by heating to boiling and then decocting for 35 to 45 minutes. In one embodiment, the decocting may be performed for 35, 38, 40, 42, or 45 minutes. The decocting state after boiling is preferably a slight boil. The resulting decoction, which does not require filtration, is Extract 2.

[0037] The extract 1 and extract 2 obtained above are mixed to obtain the Chinese medicine composition. In the present invention, after the extract 1 and extract 2 are mixed, preferably a concentration step is further included. The present invention does not specifically limit the concentration method, and conventional concentration methods in the art can be used. In one embodiment, a reduced pressure concentration method can be selected. After concentration, a thick extract is obtained, and the thick extract is preferably dried. The present invention does not specifically limit the drying method, and conventional drying methods in the art can be used. In one embodiment, a freeze-drying method can be selected to obtain a dry powder after drying.

[0038] The present invention also provides the use of the traditional Chinese medicine composition or the preparation method in preparing a drug for treating gastric precancerous lesions. The gastric precancerous lesions preferably include spasmolytic polypeptide-expressing metaplasia.

[0039] The gastric precancerous lesions preferably also include chronic atrophic gastritis. The chronic atrophic gastritis syndrome is characterized by spleen and stomach weakness, blood stasis blocking the collaterals, and symptoms include epigastric pain, fullness or distension, poor appetite, fatigue, etc.

[0040] The traditional Chinese medicine composition of the present invention is used to treat gastric precancerous lesions, can significantly alleviate the bloating symptoms and pathological changes of gastric precancerous lesions, restore chief cell-like characteristics, and significantly reduce the level of inflammatory cell infiltration; inhibit the overexpression of DMBT1, WFDC2, CFTR, AQP5 and CLU, which are markers of malignant progression of gastric precancerous lesions; reduce atrophy and intestinal metaplasia of the gastric angle and gastric antrum, effectively treat gastric precancerous lesions, and avoid the occurrence of gastric cancer.

[0041] In the pharmaceutical application of the present invention, the drug preferably further comprises a pharmaceutically acceptable excipient.

[0042] In the pharmaceutical application of the present invention, the dosage of each raw material of the Chinese medicine composition does not exceed the provisions of the pharmacopoeia standards; and the Chinese medicine composition of the present invention does not contain the 18 anti-incompatibility and 19 fear incompatibility taboos, and does not contain medicinal materials marked as toxic in the legal standards and proven to be toxic by modern toxicology.

[0043] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0044] In the following examples, unless otherwise specified, all methods are conventional.

[0045] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0046] Example 1

[0047] Take 90g of ginseng, 120g of stir-fried Atractylodes macrocephala, 150g of Astragalus membranaceus, 100g of dried ginger, 100g of dried tangerine peel, 90g of ginger pinellia, 150g of Poria, 100g of Hedyotis diffusa, 90g of vinegar-soaked Curcuma, 120g of Ganoderma lucidum, and 100g of liquorice, and mix the above materials to obtain a mixture.

[0048] The mixture was decocted in a Chinese medicine decoction bag, 10 times the amount of water (1 g mixture: 10 mL water) was added, and the mixture was soaked for 60 minutes. The mixture was heated to boiling and kept slightly boiling. After 40 minutes, the decoction was poured out. The residue was added with 10 times the amount of water (1 g mixture: 10 mL water), heated to boiling and kept slightly boiling for 40 minutes, and the decoction was poured out. The two decoctions were combined, filtered, and the filtrate was collected.

[0049] Weigh 30 g of Panax notoginseng, grind it into powder, add 16.7 times (500 mL) of water, soak for 60 minutes, heat to boiling and keep it slightly boiling, and mix it with the above filtrate after 40 minutes to obtain a mixed solution.

[0050] The mixed solution is concentrated under reduced pressure to a thick extract, and then freeze-dried to obtain a freeze-dried powder.

[0051] Example 2

[0052] Take 85g of ginseng, 125g of stir-fried Atractylodes macrocephala, 145g of Astragalus membranaceus, 95g of dried ginger, 105g of dried tangerine peel, 85g of ginger pinellia, 145g of Poria, 105g of Hedyotis diffusa, 85g of vinegar-soaked Curcuma, 115g of Ganoderma lucidum, and 95g of liquorice, and mix the above pieces evenly to obtain a mixture.

[0053] The mixture was decocted in a traditional Chinese medicine decoction bag. 8 times the amount of water was added and the mixture was soaked for 55 minutes. The mixture was heated to a boil and maintained at a simmering temperature. After 35 minutes, the decoction was discarded. The residue was added to 11 times the amount of water, heated to a boil and maintained at a simmering temperature for 42 minutes, and the decoction was discarded. The residue was added to 10 times the amount of water, heated to a boil and maintained at a simmering temperature for 44 minutes, and the decoction was discarded. The three decoctions were combined, filtered, and the filtrate was collected.

[0054] Weigh another 25 g of Panax notoginseng, grind it into powder, add 18 times (450 mL) of water, soak for 55 minutes, heat to boiling and keep it slightly boiling, and mix it with the above filtrate after 35 minutes to obtain a mixed solution.

[0055] The mixed solution is concentrated under reduced pressure to a thick extract, and then freeze-dried to obtain a freeze-dried powder.

[0056] Example 3

[0057] Take 95g of ginseng, 115g of stir-fried Atractylodes macrocephala, 155g of Astragalus membranaceus, 105g of dried ginger, 95g of dried tangerine peel, 95g of ginger Pinellia tuber, 155g of Poria, 95g of Hedyotis diffusa, 95g of vinegar-soaked Curcuma, 125g of Ganoderma lucidum, and 105g of liquorice, and mix the above pieces evenly to obtain a mixture.

[0058] The mixture was decocted in a traditional Chinese medicine decoction bag. 12 times the amount of water was added and the mixture was soaked for 65 minutes. The mixture was heated to a boil and maintained at a simmering temperature. After 45 minutes, the decoction was discarded. The residue was added to 9 times the amount of water, heated to a boil and maintained at a simmering temperature for 37 minutes, and the decoction was discarded. The residue was added to 10 times the amount of water, heated to a boil and maintained at a simmering temperature for 38 minutes, and the decoction was discarded. The residue was added to 12 times the amount of water, heated to a boil and maintained at a simmering temperature for 43 minutes, and the decoction was discarded. The four decoctions were combined, filtered, and the filtrate collected.

[0059] Weigh another 35 g of Panax notoginseng, grind it into powder, add 15 times (525 mL) of water, soak for 65 minutes, heat to boiling and keep it slightly boiling, and mix it with the above filtrate after 45 minutes to obtain a mixed solution.

[0060] The mixed solution is concentrated under reduced pressure to a thick extract, and then freeze-dried to obtain a freeze-dried powder.

[0061] Test Example 1

[0062] Identification of ingredients of the Chinese medicine composition of the present invention (referred to as the spleen-tonifying and stomach-benefiting prescription)

[0063] 1. Methods

[0064] 1.1 Preparation of test solution and reference solution

[0065] In Example 1 of the present invention, 353 g of lyophilized powder was obtained, with a yield of 28.5%. Accurately weigh 0.1 g of the lyophilized powder prepared in Example 1 (denoted as the Jianpi Yiwei Recipe lyophilized powder), add 40 mL of methanol, weigh the mass, sonicate for 30 minutes, cool to room temperature, and make up the difference. Centrifuge the supernatant at 10,000 rpm for 15 minutes to obtain the test solution.

[0066] Accurately draw appropriate amounts of deacetylcaryophyllic acid, glycyrrhizin, hesperidin, notoginsenoside R1, ginsenoside Re, ginsenoside Rg1, ginsenoside Rb1, ganoderic acid A, glycyrrhizic acid, astragaloside IV, 6-gingerol, atractylodes lactone II, atractylodes lactone I, germacrone, dehydropachymic acid, and pachymic acid reference substances, and use chromatographic methanol to prepare single reference substance stock solutions with mass concentrations of 1.24 mg / mL, 1.13 mg / mL, 0.57 mg / mL, 1.14 mg / mL, 1.64 mg / mL, 1.72 mg / mL, 2.04 mg / mL, 2.24 mg / mL, 1.80 mg / mL, 1.15 mg / mL, 2.00 mg / mL, 1.10 mg / mL, 1.11 mg / mL, 1.08 mg / mL, 1.44 mg / mL, and 1.02 mg / mL, respectively. Accurately pipette an appropriate amount of each single reference substance stock solution to prepare a mixed reference substance solution with a mass concentration of 10 μg / mL.

[0067] 1.2 Chromatographic and mass spectrometry conditions

[0068] Waters Acauity UPLC BEH C 18 Chromatographic column (100 mm × 2.1 mm, 1.7 μm); positive ion mode: gradient elution: 0.1% formic acid in acetonitrile (A) - 0.1% formic acid in water (B); negative ion mode: gradient elution: 0.01% formic acid in acetonitrile (A) - 0.01% formic acid in water (B). The gradient elution program for quantitative analysis was: 0–4.5 min, 8%–30% A; 4.5–9.5 min, 30%–70% A; 9.5–12.5 min, 70%–100% A; 12.5–14.0 min, 100% A. The gradient elution program for component analysis was: 0–2.5 min, 3% A; 2.5–18.0 min, 3%–48% A; 18.0–25.0 min, 48%–70% A; 25.0–28.0 min, 70%–100% A; and 28.0–31.0 min, 100% A. The column temperature was 50°C, the flow rate was 0.4 mL / min, and the injection volume was 5 μL.

[0069] Waters Xevo G2 Q-Tof mass spectrometer, ESI ion source, data were collected in positive and negative ion modes, capillary voltage 3.0 kV (-3.0 kV), cone voltage 40 V (-40 V), ion source temperature 120 °C, desolvation temperature 550 °C, desolvation gas flow rate 900 L / h, scan range m / z 50~1200.

[0070] The specific quantitative ion information of quantitative analysis is: acetylcarnitine (negative ion mode, MH, t R =0.79min, m / z 389.1084), liquiritin (negative ion mode, MH, t R =3.02min, m / z 417.1186), hesperidin (negative ion mode, MH, t R =3.86min, m / z 609.1819), notoginsenoside R1 (negative ion mode, M+HCOO, t R =4.45min, m / z 977.5321), ginsenoside Re (negative ion mode, M+HCOO, t R =4.71min, m / z 991.5478), ginsenoside Rg1 (negative ion mode, M+HCOO, t R =4.74min, m / z 845.4899), ginsenoside Rb1 (negative ion mode, M+HCOO, t R =6.29min, m / z 1153.6006), ganoderic acid A (negative ion mode, MH, t R =6.72min, m / z 515.3009), glycyrrhizic acid (negative ion mode, MH, t R =6.84min, m / z 821.3960), astragaloside IV (negative ion mode, M+HCOO, t R =6.89min, m / z 829.4586), 6-gingerol (negative ion mode, MH, t R =7.47min, m / z 293.1753), Atractylodes lactone II (positive ion mode, M+H, t R =8.93min, m / z 233.1542), Atractylodes lactone I (positive ion mode, M+H, t R =9.65min, m / z 231.1385), Germacron (positive ion mode, M+H, t R =10.02min, m / z 219.1749), dehydropachymic acid (negative ion mode, MH, t R =11.20min, m / z 525.3580), Pachymic acid (negative ion mode, MH, t R =11.38min, m / z 527.3736).

[0071] 2. Results

[0072] 2.1 Quantitative analysis system adaptability

[0073] Take the mixed reference solution and test solution, and inject and measure according to the conditions of "1.2". The extracted ion chromatograms of the components to be measured in positive and negative ion modes are shown in Figure 2. Figure 1 The results showed that there was no obvious interference in the determination and the method had good specificity.

[0074] 2.2 Quantitative analysis results

[0075] The test solution was prepared according to the method under "1.1" in two parallel replicates. The samples were injected, determined and the content was calculated according to the conditions under "1.2". The results showed that the contents of the index components deacetylcarnitine, liquiritin, hesperidin, notoginsenoside R1, ginsenoside Re, ginsenoside Rg1, ginsenoside Rb1, ganoderic acid A, glycyrrhizic acid, astragaloside IV, 6-gingerol, atractylodes lactone II and atractylodes lactone I in the freeze-dried powder of Jianpi Yiwei Recipe were 0.006%, 0.323%, 1.889%, 0.112%, 0.064%, 0.619%, 0.436%, 0.032%, 0.871%, 0.024%, 0.315%, 0.002% and 0.009%, respectively.

[0076] The agreed quantitative components, germacrone, dehydropachymic acid and pachymic acid, were not detected in the lyophilized powder.

[0077] Test Example 2

[0078] The therapeutic effect of the Chinese medicine composition of the present invention on gastric precancerous lesions:

[0079] Animal drug administration and modeling plan:

[0080] Forty-eight male C57BL / 6J mice were randomly divided into six groups: a control group (C), a model group (M), a positive drug group (folic acid, 1.95 mg / kg, Y), and low-dose (1.15 g / kg, GL), medium-dose (2.3 g / kg, GM), and high-dose (4.6 g / kg, GH) Jianpi Yiwei Formula groups. The Jianpi Yiwei Formula was the lyophilized powder prepared in Example 1 of the present invention. Except for the control and model groups, mice were gavaged with distilled water at a dose of 0.1 mL / 10 g. All other groups received the various doses of the drug for eight days. Starting on day 3, tamoxifen (4 g / 20 g) was administered intraperitoneally for three consecutive days. High-dose tamoxifen induces a gastric precancerous lesion model in mice, characterized by spasmolytic polypeptide-expressing metaplasia (SPEM).

[0081] On the 9th day, the mice were killed, and part of the gastric tissue was taken for pathological section observation. RNA was extracted from the other part to detect changes in the expression levels of DMBT1, GPX2, WFDC2, CFTR, AQP5 and CLU, markers of SPEM malignant progression.

[0082] Experimental results:

[0083] Pathological observation results of gastric body tissue: such as HE staining of mouse gastric body pathology ( Figure 2 ), in the tamoxifen-induced mouse SPEM gastric precancerous lesion model group (M), gastric epithelial cells shed and died, and the characteristics of the chief cells at the bottom of the gastric glands disappeared, accompanied by infiltration of inflammatory cells; in the positive drug folic acid group, the infiltration of inflammatory cells was weakened, but a large number of gastric epithelial cells were still observed to die, and the characteristics of the chief cells were not restored; the high-dose group of the Jianpi Yiwei prescription could partially restore the characteristics of the chief cells and significantly reduce the level of inflammatory cell infiltration.

[0084] SPEM malignant progression marker test results: Figure 3 As shown in the results, Jianpi Yiwei recipe can effectively reduce the overexpression of DMBT1, WFDC2, CFTR, AQP5 and CLU, the malignant progression markers of SPEM gastric precancerous lesions, induced by tamoxifen.

[0085] Test Example 3

[0086] clinical trials

[0087] 1. No adverse reactions have been reported in the clinical application of the Chinese medicine composition of the present invention. During the clinical application, 27 patients with gastric precancerous lesions who visited Shenzhen Traditional Chinese Medicine Hospital were collected and treated with the Jianpi Yiwei Recipe for 3 months (taking the mixed solution prepared in Example 1, with a daily dose of 1 / 10 of the mixed solution in Example 1, taken twice). The gastric bloating before and after treatment was scored according to the following criteria: 0: no symptoms, 1: occasional symptoms or obvious improvement, 2: alleviated symptoms, 3: severe symptoms. A paired rank sum test (Wilcoxon test) was performed before and after medication.

[0088] The results are shown in Table 1 below, which indicate that the Jianpi Yiwei prescription can effectively improve gastric distension in patients with gastric precancerous lesions.

[0089]

[0090] 2. Typical Cases

[0091] The spleen-strengthening and stomach-benefiting prescriptions used in the following cases were all mixed solutions prepared in Example 1 of the present invention. One dose was 1 / 10 of the amount of the mixed solution prepared in Example 1. The dosage was 1 dose per day, divided into two doses.

[0092] Case 1: Patient Cheng, female, 33 years old, first diagnosed on March 26, 2024. Chief complaint: intermittent upper abdominal distension for more than 3 years. Gastroscopy at an external hospital 3 years ago showed: chronic atrophic gastritis, carbon 13 breath test was positive, and the reexamination after quadruple bactericidal therapy was negative; Gastroscopy at an external hospital in 2022 showed: chronic atrophic gastritis, pathology showed: intestinal metaplasia (+), low-grade tumor-like lesions, local erosions; Gastroscopy on March 29, 2024 showed: chronic atrophic gastritis, pathology showed: chronic atrophic gastritis in the gastric antrum, mild dysplasia of individual glands, inflammatory cells (2+), atrophy (2+), intestinal metaplasia (2+), gastroscopy and pathology results are shown. Figure 4. Current symptoms: upper abdominal distension, occasional stomach pain, acid reflux, more obvious after eating, nausea, good appetite, good sleep, normal bowel movements, pale red tongue, thin white fur with teeth marks, purple bruises under the tongue, and a stringy and thin pulse. He has a history of "type 2 diabetes" for more than 5 years, with general blood sugar control, fasting blood sugar of 7.6mmol / L. He denies any history of other diseases and any family or genetic history. Western medicine diagnosis: chronic atrophic gastritis. Traditional Chinese medicine diagnosis: stomach distension, syndrome type: spleen and stomach deficiency and cold with blood stasis and depression. Treatment is to strengthen the spleen and replenish qi, activate blood circulation and benefit the stomach, supplemented by soothing the liver and regulating the spleen. I give a spleen and stomach strengthening prescription. Starting from April 2024, the "Spleen and Stomach Strengthening Prescription" decoction is taken for treatment, taken after breakfast and dinner, 1 dose per day. Advise the patient to keep a good mood, go out more and interact with people, eat regularly, avoid cold and raw food, abstain from sour, sweet and spicy food, abstain from coffee, strong tea, wine, sweet potato and honey; eat less soy products, beverages and fruits, avoid barbecue, pickled products, tobacco, alcohol and high-salt food, avoid staying up late and exercise appropriately. After 7 months, the patient's upper abdominal distension and acid reflux symptoms improved significantly. In October 2024, he went to the hospital and complained of no epigastric discomfort, no bloating and no acid reflux. In November 2024, the gastroscopy showed: chronic atrophic gastritis, and the pathology showed: chronic atrophic gastritis with erosion in the gastric antrum, mild dysplasia of local glands, inflammatory cells (2+), atrophy (1+), intestinal metaplasia (1+), and the gastroscopy and pathology results were shown. Figure 5 The patient's antral atrophy and intestinal metaplasia were improved.

[0093] Case 2: Patient Liao, female, 46 years old, first diagnosed on August 30, 2024. Chief complaint: intermittent episodes of epigastric discomfort for more than a year. Previous endoscopy on July 26, 2024 showed: superficial gastritis (referring to pathology, excluding atrophic gastritis C2), pathology showed: chronic atrophic gastritis in the gastric angle, mild dysplasia of local glands, inflammation (1+), atrophy (2+), intestinal metaplasia (1+); chronic atrophic gastritis in the gastric antrum, mild dysplasia of some glands (intestinal adenomatous dysplasia), inflammation (1+), atrophy (1+), intestinal metaplasia (2+), endoscopy and pathology results are shown in Figure 6. Current symptoms: intermittent epigastric discomfort, bloating, dull stomach pain, heartburn, hunger, no acid reflux and belching, normal appetite, good sleep, once a day bowel movement, difficult and sticky stool, usually emotionally unstable, normal menstruation, pale and dark tongue with teeth marks, slightly thick and greasy white tongue coating, purple blood stasis under the tongue, smooth pulse. Western medicine diagnosis: chronic atrophic gastritis. Traditional Chinese medicine diagnosis: epigastric pain, syndrome type: spleen and stomach deficiency and cold with blood stasis and dampness. Treatment is to strengthen the spleen and replenish qi, promote blood circulation and benefit the stomach, supplemented by soothing the liver and regulating the spleen. Give a spleen and stomach strengthening prescription. Starting from August 2024, take the "Spleen and Stomach Strengthening Prescription" decoction for treatment, take it after breakfast and dinner, one dose per day. Advise the patient to maintain a good mood, eat regularly, avoid cold, raw, spicy, barbecued, pickled foods, tobacco, alcohol, foods high in salt and oil, avoid staying up late, and exercise appropriately. Four months later, the patient's symptoms improved significantly, with no bloating, occasional epigastric discomfort, belching, acid reflux, or heartburn. A follow-up gastroscopy on December 26, 2024, showed atrophic gastritis C2 (refer to pathology). Pathology showed chronic atrophic gastritis in the gastric angle, mild reactive atypical hyperplasia of the focal glands, inflammation (1+), atrophy (1+), and intestinal metaplasia (1+). Chronic atrophic gastritis in the gastric antrum, mild atypical hyperplasia of the focal glands, inflammation (1+), atrophy (1+), and intestinal metaplasia (1+). The results of the gastroscopy and pathology are shown in the table below. Figure 7 The patient's gastric angle atrophy and antral intestinal metaplasia were improved.

[0094] Case 3: Patient Xu, female, 26 years old, first diagnosed on June 19, 2023. Main complaint: gastric distension for more than 2 years. Gastroscopy on June 5, 2023 showed: chronic atrophic gastritis (C2, refer to pathology) with bile reflux. Pathology showed: chronic atrophic gastritis in the antrum, inflammatory cells (1+), atrophy (1+); chronic atrophic gastritis in the gastric angle, inflammatory cells (1+), atrophy (1+). Gastroscopy and pathology results are shown in Figure 8 . Current symptoms: bloating, no stomach pain, belching, blood in the stool, poor appetite, good sleep, no fear of cold, pale red tongue with teeth marks and thin white fur, slightly purple bruises under the tongue, and thin pulse. Western medicine diagnosis: chronic atrophic gastritis. Traditional Chinese medicine diagnosis: stomach distension, syndrome type: spleen and stomach deficiency and cold with blood stasis and qi stagnation. Treatment is to strengthen the spleen and replenish qi, promote qi and blood circulation. Give a spleen and stomach strengthening prescription. From June 2023, take 35 doses of "Spleen and Stomach Strengthening Prescription" decoction (about 1.5 months), take after breakfast and dinner, 1 dose per day. Advise the patient to keep a good mood, eat regularly, avoid cold, raw, spicy, barbecued, pickled products, tobacco, alcohol, high-salt and high-oil foods, avoid staying up late, and exercise appropriately. After more than a month, the patient's symptoms improved significantly. He reported no bloating, no stomach pain, occasional belching, no blood in his stool, good appetite, normal sleep, no fear of cold, pale red tongue with light teeth marks and thin white fur, slightly purple sublingual veins, and a thready pulse. The medication was then discontinued. One year later, on August 3, 2024, a follow-up gastroscopy showed: chronic atrophic gastritis (C1, refer to pathology). Pathology showed: chronic atrophic gastritis of the gastric antrum, inflammation (mild), atrophy (mild), and intestinal metaplasia (mild). The gastroscopy and pathology results are shown in the figure below. Figure 9 Because no obvious abnormalities were found in the gastric angle, no pathological examination was performed on the gastric angle. The patient's atrophy and intestinal metaplasia of the gastric angle and gastric antrum were significantly improved.

[0095] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. Use of a traditional Chinese medicine composition in the preparation of a drug for treating gastric precancerous lesions, characterized in that: The composition is composed of the following raw materials in parts by weight: 85-95 parts of ginseng, 115-125 parts of stir-fried atractylodes macrocephala, 145-155 parts of astragalus, 25-35 parts of notoginseng, 95-105 parts of dried ginger, 95-105 parts of dried tangerine peel, 85-95 parts of ginger pinellia, 145-155 parts of poria, 95-105 parts of oldenlandia diffusa, 85-95 parts of vinegared zedoaria, 115-125 parts of ganoderma lucidum, and 95-105 parts of liquorice; The preparation method of the traditional Chinese medicine composition comprises the following steps: mixing ginseng, stir-fried atractylodes macrocephala, astragalus root, dried ginger, tangerine peel, ginger pinellia, poria, hedyotis diffusa, vinegared zedoaria, ganoderma lucidum and liquorice to obtain a mixture, mixing the mixture with water, soaking the mixture, and decocting and extracting to obtain extract 1; Grind Panax notoginseng, mix with water, soak, and boil to obtain extract 2; Mixing extract 1 and extract 2 to obtain the traditional Chinese medicine composition; The gastric precancerous lesions include spasmolytic polypeptide-expressing metaplasia and chronic atrophic gastritis.

2. The use according to claim 1, characterized in that The traditional Chinese medicine composition is composed of the following raw materials in parts by weight: 88-92 parts of ginseng, 118-122 parts of stir-fried atractylodes macrocephala, 148-152 parts of astragalus, 28-32 parts of panax notoginseng, 98-102 parts of dried ginger, 98-102 parts of tangerine peel, 88-92 parts of ginger pinellia, 148-152 parts of poria, 98-102 parts of oldenlandia diffusa, 88-92 parts of vinegared zedoaria, 118-122 parts of ganoderma lucidum and 98-102 parts of liquorice.

3. The use according to claim 1, characterized in that The mass volume ratio of the mixture to water is 1 g: (8-12) mL, and the mass volume ratio of Panax notoginseng to water is 1 g: (15-18) mL.

4. The use according to claim 1, characterized in that The soaking time is 55 to 65 minutes.

5. The use according to claim 1, characterized in that When extract 1 is obtained, the number of decoction extractions is ≥ 2 times.

6. The use according to claim 1, characterized in that The decoction extraction comprises heating to boiling and then decocting for 35 to 45 minutes.

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