A composition for treating chronic gastritis and use thereof

Abstract

The present application relates to the field of traditional Chinese medicine, and particularly to a composition for treating chronic gastritis and application thereof, the composition is prepared from the following raw materials according to weight fraction: 8-12 parts of Fashanxia, 8-12 parts of Huangqin, 3-7 parts of Huanglian, 4-8 parts of Ganjiang, 12-18 parts of Taizishen, 12-18 parts of Baihuasheshecao, 12-16 parts of Banzhilian, 8-12 parts of E'zhu, 12-20 parts of Chaozhigu, 18-22 parts of Chaoquya, 18-22 parts of Chaomaiya, 8-12 parts of Yujin, 1-5 parts of Sanqi, 8-12 parts of Dazao and 3-8 parts of Zhigancao. The present application effectively solves the problems of lack of specific therapy and specific intervention target for reversing mucosal atrophy, intestinal metaplasia and other precancerous lesions in the prior art, provides a new idea for the mechanism research of CAG treatment by traditional Chinese medicine, and has important theoretical value and clinical conversion prospect for promoting the modernization of traditional Chinese medicine.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine, specifically to a composition for treating chronic gastritis and its application. Background Technology

[0002] Chronic atrophic gastritis (CAG) has a high clinical prevalence, a long course, and often nonspecific symptoms. Gastric cancer (GC) is prevalent globally and is one of the leading causes of cancer death. In 1975, Correa et al. proposed the classic evolutionary model of gastric cancer: chronic superficial gastritis → CAG → intestinal metaplasia (IM) → dysplasia → GC. CAG is a crucial link in the gastric inflammation-cancer transformation process and a key stage for drug intervention. Furthermore, surveys show that the incidence of GC is as high as 6% in individuals with CAG accompanied by IM and dysplasia. Therefore, effective prevention and treatment of CAG is of great significance in reducing the incidence and mortality of GC.

[0003] Current clinical intervention strategies for gastric atrophy (CAG) face significant challenges. Traditional treatments primarily include Helicobacter pylori eradication therapy, acid suppression and gastric protection, antioxidant therapy, and regulation of gastrointestinal function. While these interventions can partially alleviate symptoms or reverse mild atrophy, their effectiveness in reversing pathological changes and effectively reducing the risk of cancer remains uncertain for patients with moderate to severe CAG, accompanied by extensive intramucosal metaplasia (IM) or dysplasia. The fundamental reason lies in the incomplete understanding of the deep mechanisms underlying the complex biological process of gastric inflammation-cancer transformation, and the lack of specific therapies and intervention targets for reversing precancerous lesions such as mucosal atrophy and intestinal metaplasia. Therefore, elucidating the intrinsic molecular mechanisms of CAG malignant transformation and finding safe and effective strategies for multi-target, multi-stage intervention are urgent scientific frontiers and clinical needs in the field of gastric cancer prevention and treatment. Summary of the Invention

[0004] In view of the shortcomings and deficiencies of the existing technology, the purpose of this invention is to provide a traditional Chinese medicine composition for treating chronic gastritis, reveal the molecular mechanism by which the composition effectively intervenes in the "inflammation-cancer" transformation of CAG, provide new ideas for the study of the mechanism of traditional Chinese medicine treatment of CAG, and have important theoretical value and clinical translation prospects for promoting the modernization of traditional Chinese medicine.

[0005] To achieve the objective of this invention, the inventors provide the following technical solution: This invention first provides a composition for treating chronic gastritis—hereinafter referred to as "Xiaoliu Zhiwei Formula" or "XLZWF" for ease of reference—which is prepared from the following raw materials in parts by weight: 8-12 parts of Pinellia ternata, 8-12 parts of Scutellaria baicalensis, 3-7 parts of Coptis chinensis, 4-8 parts of Zingiber officinale, 12-18 parts of Codonopsis pilosula, 12-18 parts of Hedyotis diffusa, 12-16 parts of Scutellaria barbata, 8-12 parts of Curcuma zedoaria, 12-20 parts of Citrus aurantium (fried), 18-22 parts of germinated barley (fried), 18-22 parts of malt (fried), 8-12 parts of Curcuma longa, 1-5 parts of Panax notoginseng, 8-12 parts of Ziziphus jujuba, and 3-8 parts of Glycyrrhiza uralensis (processed).

[0006] As a preferred embodiment, the composition for treating chronic gastritis according to the present invention is prepared from the following raw materials in parts by weight: 10 parts of Pinellia ternata, 10 parts of Scutellaria baicalensis, 5 parts of Coptis chinensis, 6 parts of dried ginger, 15 parts of Codonopsis pilosula, 15 parts of Hedyotis diffusa, 15 parts of Scutellaria barbata, 10 parts of Curcuma zedoaria, 15 parts of stir-fried Citrus aurantium, 20 parts of stir-fried germinated barley, 20 parts of stir-fried malt, 10 parts of Curcuma longa, 3 parts of Panax notoginseng, 10 parts of jujube, and 6 parts of prepared licorice root.

[0007] The "tumor-eliminating and atrophy-treating formula" described in this invention is an empirical formula obtained by the inventor's research team under the guidance of Professor Zhang Xiaoping, a nationally renowned traditional Chinese medicine doctor, on the "theory of spleen and stomach qi transformation," combined with years of clinical exploration and practice.

[0008] Professor Zhang Xiaoping believes that the transformation of CAG from inflammation to cancer is a dynamic evolutionary process of "qi stagnation - turbidity accumulation - toxic stasis - atrophy". The pathogenesis is rooted in spleen and stomach weakness, with deficiency of vital energy and stagnation of pathogenic factors, leading to the generation of pathological products such as damp-heat and phlegm stasis, forming a pathogenesis of deficiency in the root and excess in the branch, and a complex mixture of cold and heat. She proposes the treatment principle of "balancing cold and heat, regulating ascending and descending functions", combined with regulating qi and blood circulation, eliminating stasis and detoxifying, removing atrophy and promoting new tissue growth, and restoring the ascending and descending functions of the spleen and stomach. Clinically, she modified the ancient formula Banxia Xiexin Tang to create a formula for eliminating tumors and treating atrophy. This formula, based on Banxia Xiexin Tang, adds the detoxifying, antibacterial, and anti-inflammatory herbs Hedyotis diffusa and Scutellaria barbata; the qi-regulating, stagnation-dispersing, blood-activating, and new tissue-generating herbs Curcuma zedoaria, Panax notoginseng, and Curcuma longa; and the qi-regulating herbs of stir-fried Citrus aurantium, stir-fried germinated rice, and stir-fried malt. The entire formula uses both cold and heat herbs, regulating both qi and blood. Figure 1 Experiments have shown that the tumor-eliminating and atrophy-treating formula of the present invention is significantly effective in treating CAG and precancerous lesions.

[0009] Preferably, in the composition for treating chronic gastritis according to the present invention, the composition is a pharmaceutical preparation selected from: decoctions, tablets, capsules, lozenges, granules, pills, powders, ointments, elixirs, suspensions, solutions, injections, suppositories, ointments, sprays, drops, drop pills, or patches. Further, more preferably, the pharmaceutical preparation of the composition is an ointment.

[0010] As a preferred embodiment, according to the present invention, a composition for treating chronic gastritis comprises the following ingredients: Pinellia ternata 10g, Scutellaria baicalensis 10g, Coptis chinensis 5g, Zingiber officinale 6g, Codonopsis pilosula 15g, Hedyotis diffusa 15g, Scutellaria barbata 15g, Curcuma zedoaria 10g, Citrus aurantium 15g, Hordeum vulgare 20g, Hordeum vulgare 20g, Curcuma longa 10g, Panax notoginseng 3g, Ziziphus jujuba 10g, and Glycyrrhiza uralensis 6g, prepared as a paste. It is taken twice daily, half an hour after breakfast and dinner, one packet each time, dissolved in a small amount of warm water.

[0011] The above provides a composition for treating chronic gastritis. To make the inventive objectives, composition features, and advantages of this invention more apparent and understandable, the invention will be further described in detail below with reference to specific embodiments.

[0012] The present invention also provides the use of the above composition in the preparation of a medicament for treating chronic gastritis; further, the use in the preparation of a medicament for treating chronic atrophic gastritis is preferred.

[0013] The present invention also provides the application of the above composition in the potential regulation of cellular homeostasis through multi-pathway synergy. Preferably, the multi-pathway synergy includes the PI3K / AKT / P53 signaling pathway and autophagy-related pathways.

[0014] Experiments show that the tumor-eliminating and atrophy-treating formula of the present invention may exert its therapeutic effect by inhibiting the PI3K / AKT / P53 pathway to activate cell autophagy, and by synergistically exerting anti-inflammatory, anti-aging and gastric mucosal function effects.

[0015] Experiments show that the tumor-reducing and atrophy-treating formula of the present invention may play a role in reversing IM by inhibiting the PI3K / AKT / P53 pathway and activating autophagy.

[0016] Compared with the prior art, the beneficial effects of the present invention are: (1) Chronic atrophic gastritis (CAG) is a key stage of precancerous lesions of the stomach. Effectively inhibiting or reversing the progression of CAG is of great significance for preventing the occurrence of gastric cancer. This invention, by observing the effects of Xiaoliu Zhiwei Decoction on gastric mucosal pathology, aging phenotype, mitochondrial function and autophagy and related indicators of CAG rats after intervention, and its correlation with the p53 / PINK1 / Parkin signaling axis, can innovatively reveal the molecular mechanism by which Xiaoliu Zhiwei Decoction effectively intervenes in the "inflammation-cancer" transformation of CAG. This provides a new idea for the study of the mechanism of traditional Chinese medicine in treating CAG, which is in line with the TCM diagnosis and treatment concept of "preventing disease before it occurs and preventing the progression of disease after it occurs". It enriches the theoretical connotation of TCM "treating disease before it occurs" and is conducive to the research and development of new TCM compound prescriptions. It has high clinical application value.

[0017] (2) Chronic atrophic gastritis (CAG) is a critical precancerous stage in the development of gastric cancer, and there is a lack of effective interventions in clinical practice. The tumor-eliminating and atrophic formula developed by the research team of this invention can effectively improve gastric mucosal atrophy and intestinal metaplasia. The research group has previously confirmed through in vitro and in vivo experiments that the tumor-eliminating and atrophic formula can inhibit the activation of cell autophagy by the p53 pathway and exert therapeutic effects in synergistic anti-inflammatory, anti-aging, and improvement of gastric mucosal function. Based on the bidirectional regulation of the PINK1 / Parkin pathway by p53 and the core role of mitophagy in clearing senescent cells, the inventors proposed the hypothesis that the tumor-eliminating and atrophic formula regulates mitophagy through the p53 / PINK1 / Parkin signaling axis to resist cell senescence and intervene in the "inflammation-cancer" transformation of CAG. This invention aims to use a multi-factor combined method to induce a CAG rat model and an doxorubicin-induced GES-1 cell senescence model. Through strategies such as dynamic monitoring of mitochondrial autophagy, detection of cell senescence phenotypes, gene silencing and overexpression, and single-cell transcriptome sequencing, the study will verify the mechanism by which the Xiaoliu Zhiwei formula regulates mitochondrial function and alleviates SASP through the p53 / PINK1 / Parkin pathway, thereby blocking the "inflammation-cancer" transformation of CAG. This will provide some scientific basis for the prevention and treatment of precancerous lesions of gastric cancer using traditional Chinese medicine. Attached Figure Description

[0018] Figure 1 This is a schematic diagram of the "tumor-eliminating and atrophy-treating formula" of the present invention.

[0019] Figure 2 These are heatmaps and volcano plots of DEGs identified in the GEO dataset of Experiment Example 1 of this invention, and heatmaps and volcano plots of the first 25 genes and DEGs in the gastritis and LGIN groups in the GSE130823 dataset.

[0020] Figure 3 These are heatmaps and volcano plots of DEGs identified in the GEO dataset of Experiment Example 1 of this invention, and heatmaps of the first 25 genes and DEGs in the gastritis and HGIN groups in the GSE130823 dataset.

[0021] Figure 4 These are heatmaps and volcano plots of DEGs identified in the GEO dataset of Experiment Example 1 of this invention, and heatmaps and volcano plots of the first 25 genes in the gastritis and IGC groups in the GSE130823 dataset.

[0022] Figure 5 These are heatmaps and volcano plots of DEGs identified in the GEO dataset of Experiment Example 1 of this invention, and heatmaps and volcano plots of the first 25 genes and DEGs in the chronic gastritis and LGIN groups in the GSE55696 dataset.

[0023] Figure 6These are heatmaps and volcano plots of DEGs identified in the GEO dataset of Experiment Example 1 of this invention, and heatmaps and volcano plots of the first 25 genes and DEGs in the chronic gastritis and HGIN groups in the GSE55696 dataset.

[0024] Figure 7 These are heatmaps and volcano plots of DEGs identified in the GEO dataset of Experiment Example 1 of this invention, and heatmaps and volcano plots of the first 25 genes in the chronic gastritis and EGC groups in the GSE55696 dataset.

[0025] Figure 8 This is the diagram from Experimental Example 1 of this invention. Wherein: ① is a schematic diagram of the intersection target sites of XLZWF and CAG genes; ② is a GO functional enrichment analysis diagram; ③ is a KEGG pathway enrichment analysis diagram.

[0026] Figure 9 This is the spectrum from Experimental Example 1 of this invention. Specifically: ① is a schematic diagram showing 56 key targets identified through screening of XLZWF's blood-entry components; ② is a schematic diagram showing the molecular docking results.

[0027] Figure 10 This shows the gastric mucosa and pathological changes in CAG rats in Experiment Example 2 of this invention. Wherein: A represents macroscopic examination photographs of the stomachs of rats in different groups.

[0028] Figure 11 This describes the gastric mucosa and pathological changes in CAG rats in Experiment Example 2 of this invention. Specifically: B is a typical representative section showing the pathological characteristics of rat gastric tissue stained with HE; C is a typical representative section showing the pathological characteristics of rat gastric tissue stained with Masson's Law.

[0029] Figure 12 This describes the serum inflammatory markers of different groups of CAG rats in Experiment Example 2 of this invention. Wherein: D and E represent the serum levels of PG I and PG II, respectively; F and G represent the PG I / PG II ratio and the serum level of GAS-17, respectively; and H, I, and J represent the serum levels of IL-1β, IL-6, and TNF-α, respectively. Results are expressed as mean ± standard deviation (n = 6). ### p<0.001 vs. control group; ns p>0.05, * p<0.05, ** p<0.01, *** p<0.001 vs. model group.

[0030] Figure 13This describes the effect of XLZWF on the PI3K / AKT / P53 signaling pathway in vivo, as shown in Experimental Example 3 of this invention. Wherein: A, B, and C represent the mRNA expression levels of PI3K, AKT, and P53 in gastric tissue, respectively; D, E, and F represent the protein expression levels of PI3K, phosphorylated AKT (p-AKT), and AKT, respectively; G and H represent the protein expression levels of p-AKT and p53, respectively. Results are expressed as mean ± standard deviation or M (P25, P75). n = 6. ### p<0.001 vs control group; ns p>0.05, * p<0.05, ** p<0.01, *** p<0.001 vs model group.

[0031] Figure 14 This describes the effect of XLZWF on autophagy-related factors in vivo, as shown in Experimental Example 3 of this invention. Wherein: I is a representative Western blot image; J, K, and L represent the mRNA expression of autophagy-related markers LC3, Beclin-1, and Atg13, respectively; M, N, and O represent the protein expression of autophagy-related markers LC3, Beclin-1, and Atg13, respectively. Results are expressed as mean ± standard deviation or M (P25, P75). n = 6. ### p<0.001 vs control group; ns p>0.05, * p<0.05, ** p<0.01, *** p<0.001 vs model group.

[0032] Figure 15 This is a diagram illustrating the process of establishing the PLGC cell model in Experiment Example 4 of this invention. A shows cell survival at different MNNG concentrations, used to determine a safe concentration range suitable for in vitro use; B, C, and D show the protein expression of KLF4, MUC2, and VILLIN at a 40 μM MNNG dose, respectively; E shows the mRNA expression of KLF4 at a 40 μM MNNG dose. Results are expressed as mean ± standard deviation. ns p>0.05, ## p<0.01, ### p<0.00 vs control group; ** p<0.01 vs model group.

[0033] Figure 16This diagram illustrates the process of establishing the PLGC cell model in Experiment Example 4 of this invention and the effect of XLZWF-containing serum. Specifically: F and G show the mRNA expression of MUC2 and VILLIN at a dose of 40 μM MNNG, respectively; H is a representative Western blot image; I shows cell survival at different concentrations of XLZWF-containing serum, used to determine the optimal in vitro drug concentration; J shows the effect of XLZWF-containing serum on the survival rate of PLGC model cells, expressed as mean ± standard deviation. ns p>0.05, ## p<0.01, ### p<0.00 vs control group; ** p<0.01 vs model group.

[0034] Figure 17 This describes the effect of XLZWF-containing serum on the results of Experimental Example 4 of this invention. Wherein: A is a representative Western blot image; BG bar graph shows the relative mRNA and protein expression of KLF4, MUC2, and VILLIN in PLGC cells. Results are expressed as mean ± standard deviation. # p<0.05, ## p<0.01, ### p<0.001 vs control group; ** p<0.01, *** p<0.001 vs model group.

[0035] Figure 18 This describes the effect of XLZWF-containing serum on the results of Experimental Example 4 of this invention. Wherein: H, I, J, and K represent the mRNA expression levels of PI3K, AKT, mTOR, and p53, respectively; L, M, N, and O represent the protein expression levels of PI3K, p-PI3K, AKT, and p-AKT, respectively; P, Q, and R represent the protein expression levels of mTOR, p-mTOR, and p53, respectively. Results are expressed as mean ± standard deviation. # p<0.05, ## p<0.01, ### p<0.001 vs control group; ** p<0.01, *** p<0.001 vs model group.

[0036] Figure 19 This describes the effect of XLZWF-containing serum on the results of Experiment 4 of this invention. S, T, U, and V represent the mRNA and protein expression levels of Beclin-1 and LC3, respectively. Results are expressed as mean ± standard deviation. # p<0.05, ## p<0.01,### p<0.001 vs control group; ** p<0.01, *** p<0.001 vs model group.

[0037] Figure 20 This describes the effect of XLZWF on aging-related phenotypes in CAG rats in Experimental Example 5 of this invention. Wherein: A is a representative Western blot image; B, C, and D show the protein expression levels of VEGFR1, VEGFR2, and COX-2 in each group; E, F, and G show the mRNA expression levels of VEGFR1, VEGFR2, and COX-2 in each group. Detailed Implementation

[0038] The present invention will be described in more detail below with reference to the embodiments. It should be understood that the implementation of the present invention is not limited to the embodiments below, and any modifications and / or alterations made to the present invention will fall within the protection scope of the present invention.

[0039] In this invention, unless otherwise specified, all parts and percentages are by weight, and all equipment and raw materials are commercially available or commonly used in the industry. Unless otherwise specified, the methods used in the embodiments are general techniques in the art.

[0040] Example 1 A composition for treating chronic gastritis, known as "Xiaoliu Zhiwei Fang," comprises, by weight: 10g of prepared Pinellia ternata, 10g of Scutellaria baicalensis, 5g of Coptis chinensis, 6g of dried ginger, 15g of Codonopsis pilosula, 15g of Hedyotis diffusa, 15g of Scutellaria barbata, 10g of Curcuma zedoaria, 15g of stir-fried Citrus aurantium, 20g of stir-fried germinated barley, 20g of stir-fried malt, 10g of Curcuma longa, 3g of Panax notoginseng, 10g of jujube, and 6g of prepared licorice root.

[0041] The relevant experimental results are shown in the experimental examples section below.

[0042] Example 2 A composition for treating chronic gastritis, known as "Xiaoliu Zhiwei Fang," comprises, by weight: 8g of Pinellia ternata, 8g of Scutellaria baicalensis, 3g of Coptis chinensis, 5g of Zingiber officinale, 12g of Codonopsis pilosula, 12g of Hedyotis diffusa, 12g of Scutellaria barbata, 10g of Curcuma zedoaria, 13g of Citrus aurantium (fried), 18g of germinated barley, 22g of malt, 8g of Curcuma longa, 2g of Panax notoginseng, 8g of Ziziphus jujuba, and 4g of Glycyrrhiza uralensis (processed).

[0043] The experimental results are similar to those in Example 1, and will not be repeated here.

[0044] Example 3 A composition for treating chronic gastritis, known as "Xiaoliu Zhiwei Fang," comprises, by weight: 12g of prepared Pinellia ternata, 12g of Scutellaria baicalensis, 4g of Coptis chinensis, 8g of dried ginger, 18g of Codonopsis pilosula, 18g of Hedyotis diffusa, 16g of Scutellaria barbata, 12g of Curcuma zedoaria, 20g of stir-fried Citrus aurantium, 22g of stir-fried germinated barley, 18g of stir-fried malt, 12g of Curcuma longa, 5g of Panax notoginseng, 12g of jujube, and 8g of prepared licorice root.

[0045] The experimental results are similar to those in Example 1, and will not be repeated here.

[0046] Example 4 A composition for treating chronic gastritis, known as "Xiaoliu Zhiwei Fang," comprises, by weight: 10g of prepared Pinellia ternata, 12g of Scutellaria baicalensis, 7g of Coptis chinensis, 4g of dried ginger, 16g of Codonopsis pilosula, 16g of Hedyotis diffusa, 12g of Scutellaria barbata, 8g of Curcuma zedoaria, 18g of stir-fried Citrus aurantium, 20g of stir-fried germinated barley, 18g of stir-fried malt, 10g of Curcuma longa, 5g of Panax notoginseng, 12g of jujube, and 3g of prepared licorice root.

[0047] The experimental results are similar to those in Example 1, and will not be repeated here.

[0048] Experimental Examples Experimental Example 1: Analysis of the Effective Components of the Tumor-Eliminating and Atrophy-Treating Formula Sample preparation: Ten SPF-grade male SD rats were randomly divided into a control group and an XLZWF group. The XLZWF group was administered 3.686 g / kg XLZWF suspension by gavage, while the control group was administered 2 mL of physiological saline by gavage daily for 7 consecutive days. Rats were fasted for 12 h before the last administration. One h after administration, rats were anesthetized (2% sodium pentobarbital, 0.2 mL / 100 g), and blood was collected from the abdominal aorta in heparin sodium anticoagulant tubes. The blood was incubated at 4℃ for 2 h, centrifuged at 3000 r / min for 15 min, and the supernatant was collected. The intact liver of the rats was harvested, rinsed with pre-cooled physiological saline or PBS, blotted dry with filter paper, and approximately 100 mg of liver tissue was immediately flash-frozen in liquid nitrogen. All liver tissue was stored at -80 ℃ for later use.

[0049] ① XLZWF aqueous extract: Accurately measure 100 μL of XLZWF suspension, dilute with methanol to a final volume, vortex to mix, and then sonicate in an ice bath to ensure complete extraction. Subsequently, centrifuge at high speed, collect the supernatant, filter through a 0.22 μm organic filter membrane, and collect the filtrate.

[0050] ② Serum sample extraction: Take 100 μL of rat plasma supernatant, precipitate proteins with acetonitrile, vortex, centrifuge, take the supernatant, blow dry with nitrogen at room temperature, and redissolve with acetonitrile.

[0051] ③ Liver tissue sample extraction: After grinding the liver sample with liquid nitrogen, 50 mg was added to 1 mL of pre-cooled extraction solution, homogenized in an ice bath, and ultrasonically broken up. The homogenate was incubated at -20℃ for 1 hour, centrifuged, and the supernatant was collected. After vacuum drying, it was reconstituted with 100 μL of acetonitrile:water (1:1) for subsequent analysis.

[0052] Ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HDMS) was used to analyze and identify the aqueous extract of the Xiaoliu Zhiwei formula, the serum containing the Xiaoliu Zhiwei formula, and blank serum. A total of 465 chemical components were identified in the liver tissue of rats in the XLZWF group (280 in positive ion mode and 185 in negative ion mode). A total of 1024 chemical components were identified in the serum of rats containing XLZWF (557 in positive ion mode and 467 in negative ion mode). Comparison between blood-entering components and liver metabolic components revealed that serum components were mainly composed of the original drug and its primary conjugates (such as glucuronic acid / sulfate conjugates), reflecting the systemic exposure and transport of the drug. Liver components, however, exhibited a significantly different metabolic profile, characterized by abundant phase I (hydroxylation, demethylation) and phase II (conjugation) metabolites, accompanied by extensive reprogramming of endogenous metabolites (such as bile acids and fatty acids). Comparative analysis with the chemical component spectra in the aqueous extract of the traditional Chinese medicine and the blank rat serum identified 129 original blood-entering components.

[0053] To explore the potential biological functions of the active ingredients in the Xiaoliu Zhiwei formula, we searched for its pharmacodynamic targets in 129 blood-entering components of the formula using databases such as SwissTargetPrediction, STITCH, SEA, and TCMSP. After deduplication, a total of 2237 pharmacodynamic targets were obtained. 750 CAG disease targets were obtained using the GSE130823 dataset. Figure 2-4 The GSE55696 dataset yielded 822 disease targets for CAG. Figure 5-7 The Genecard database yielded 967 CAG disease targets, which, after deduplication, resulted in 2233 CAG disease targets. Correlation was established between these disease targets and the efficacy targets of the anti-tumor and anti-atrophy formula, identifying 365 common targets. Figure 8 ① in the middle.

[0054] GO functional enrichment analysis suggests that the biological processes targeting blood-entering components are mainly enriched in key functions such as apoptosis inhibition, inflammatory response, growth factor signaling pathways (e.g., EGFR / IGF), hypoxia response, and cell proliferation regulation. In terms of cellular components, they are mainly located in structures such as the cell membrane, extracellular region, exosomes, and membrane receptor complexes. Molecularly, they are characterized by protein interactions (such as homologous binding and dimerization), kinase activity, and enzyme binding, involving biochemical activities such as signal transduction, enzymatic regulation, and epigenetic modification. Figure 8 (② in the middle).

[0055] KEGG pathway enrichment analysis revealed that the target sites of the blood-entered components play important roles in key biological processes such as tumorigenesis (e.g., cancer pathways, prostate cancer), metabolic and vascular diseases (e.g., atherosclerosis), cell signaling and apoptosis regulation (e.g., PI3K / AKT, P53 pathways), and infection immunity and autophagy (e.g., cytomegalovirus infection, autophagy pathway). In particular, they were significantly enriched in core pathways closely related to cellular homeostasis, such as PI3K / AKT, P53, and autophagy. Figure 8 (③ in the middle).

[0056] The target proteins were uploaded to the STRING database to obtain the PPI network. Cluster analysis of the PPI protein network was performed using the MCODE plugin in Cytoscape 3.10.1 software (all settings were default), and the data was sorted by degree value, resulting in 56 core targets including AKT1, TP53, EGFR, KRAS, HRAS, BCL2, BCL2L1, CASP3, CASP8, CASP9, and HIF1A. Figure 9 (① in Table 1).

[0057] Meanwhile, using Cytoscape 3.10.1 software, an XLZWF-component-CAG-target network diagram was constructed and sorted from high to low degree values ​​to screen out core active ingredients such as isoscoparin, cimicifugoside, 4-hydroxybenzoic acid, vanillin, and H1,2,4-hydroxyhydroquinone.

[0058] Table 1. Protein density information of core target sites The main active ingredients, isoscoparin, cimicifugoside, 4-hydroxybenzoic acid, vanillin, and hydroxyhydroquinone, were molecularly docked with their target proteins (AKT1, TP53, BCL2, BCL2L1, and HIF1A). Figure 9 (②, AI). The results showed that most of the active ingredients had good binding activity with the target protein.

[0059] Mass spectrometry analysis identified 1024 and 465 chemical components in the serum and liver of drug-treated rats, respectively, of which 129 were the original components entering the bloodstream, indicating that the drug undergoes extensive metabolic activation in vivo. Network pharmacology analysis further identified 365 key targets, and enrichment analysis showed that these targets were significantly enriched in biological processes such as apoptosis regulation, inflammatory response, PI3K / AKT / P53 signaling pathway, and autophagy, suggesting that XLZWF may regulate cellular homeostasis through multiple pathways. PPI network screening identified 56 core targets, including AKT1, TP53, and EGFR, and identified core active ingredients such as isochrysine and cimicifugain. Finally, molecular docking verified the stable binding between the above components and key target proteins.

[0060] Experimental Example 2: The tumor-eliminating and atrophy-treating formula improves pathological damage and gastric function in PL-CAG rats by inhibiting inflammation. Multifactorial composite modeling method: A CAG rat model was established by allowing free access to MNNG solution, administering ranitidine combined with alcohol via gavage, and simultaneously inducing starvation / fullness disorders. Rats in the model group received free access to 150 mg / L MNNG solution daily and were given ranitidine (0.03 g / kg) via gavage daily. Simultaneously, a starvation / fullness disorder was implemented (2 days of feeding followed by 1 day of fasting). On the morning of the first day of the feeding cycle, rats were administered 56℃, 150 g / L NaCl solution (2 mL / 100 g) via gavage, and on the afternoon of the fasting cycle, they were administered 30% ethanol via gavage. The model was established for 12 consecutive weeks. (Related content can be found in the following references: [1] Yu Chunyue, Liu Ting, Liu Kaige, et al. Establishment and evaluation of animal model of chronic atrophic gastritis syndrome [J]. Chinese Journal of Traditional Chinese Medicine, 2018, 33(05): 2140-4; [2] Chen Xinyi, Shu Jin. Research progress on rat model of gastric precancerous lesions induced by MNNG solution combined with multifactors [J]. Chinese Journal of Comparative Medicine, 2022, 32(08):104-8; [3] Zhuang Kunhai. Study on intervention mechanism of spleen-tonifying, heat-clearing and blood-activating method for chronic atrophic gastritis based on "inflammation-cancer chain" [D]; Guangzhou University of Traditional Chinese Medicine, 2017.) Male SPF-grade SD rats with a weight range of 200±20 g were used in the experiment. After modeling, rats (n=30) were randomly divided into five groups using a random number table: Model group, low-dose XLZWF group (XLZWF-L), medium-dose XLZWF group (XLZWF-M), high-dose XLZWF group (XLZWF-H), and Weifuchun group (WFC), with six rats in each group. The equivalent drug dose was calculated using the human-rat surface area conversion formula, and referring to the conversion method for animal model drug doses in "Animal Models of Human Diseases," XLZWF low, medium, and high doses were determined to be 0.5 times, 1 times, and 2 times the clinical equivalent dose, respectively. The XLZWF-L, XLZWF-M, and XLZWF-H groups were administered XLZWF suspension by gavage at doses of 1.843, 3.686, and 7.372 g / kg, respectively. The WFC group was administered the adult equivalent dose (0.441 g / kg) by gavage. The control and model groups were administered 2 ml of normal saline via gavage once daily for 12 consecutive weeks.

[0061] In the previous study, the research group used a multi-factor composite modeling method to establish a CAG rat model. After successful modeling, the rats were randomly divided into five groups: the model group, the low-dose group of Xiaoliu Zhiwei Fang (L-XLZWF), the medium-dose group of Xiaoliu Zhiwei Fang (M-XLZWF), the high-dose group of Xiaoliu Zhiwei Fang (H-XLZWF), and the Weifuchun group (WFC). After 12 weeks of continuous administration, blood and tissue samples were collected from the rats. The efficacy was evaluated by HE and Masson staining of the gastric mucosa tissue. The levels of PG I, PG II, PGR (PG I / PG II), GAS-17, IL-1β, TNF-α, and IL-6 in the rat serum were detected. The results are as follows: ①Gastric mucosa and pathological changes in each group of rats Visual observation: At week 24, compared with the control group, the model group had a smaller stomach volume and a lighter color, indicating a thinner mucosal layer and fewer folds. In all treatment groups, the stomach volume increased, the mucosal layer was thicker, and the folds were more obvious during visual examination. Figure 10 (A in the middle).

[0062] HE staining showed that at week 24, compared with the control group, the gastric mucosa in the model group was structurally disrupted and disordered, with extensive necrosis of epithelial cells and structural disorder. The number of glands was reduced, and atrophy was observed. Extensive diffuse inflammatory cell infiltration was present in the lamina propria of the mucosa. Compared with the model group, the XLZWF-L group showed milder gastric mucosal damage, with epithelial cell shedding in some areas, moderate inflammatory cell infiltration, and some recovery of glandular structure, although some atrophy remained. The XLZWF-M group showed mostly restored gastric mucosal structure, relatively orderly arrangement of epithelial cells, reduced inflammatory cell infiltration to a mild level, and effective inhibition of glandular atrophy. The XLZWF-H group showed relatively intact gastric mucosal structure, more orderly arrangement of epithelial cells, significantly reduced inflammatory cell infiltration, and some recovery of glandular structure. A small number of scattered inflammatory cell infiltrations were observed. Figure 11 (B in the middle).

[0063] Masson staining showed that at week 24, compared with the control group, the model group exhibited disordered gastric mucosal structure, extensive inflammatory cell infiltration, and numerous coarse blue collagen fibers in the interstitium, distributed in a reticular or sheet-like pattern, encapsulating glands. Compared with the model group, the XLZWF-L group showed less significant recovery of gastric mucosal structure, a slight reduction in inflammatory cell infiltration, and a slight decrease in the area and density of blue collagen fibers compared to the model group; the XLZWF-M group showed varying degrees of recovery of gastric mucosal structure, a slight reduction in inflammatory cell infiltration, and a decrease in the area and density of blue collagen fibers compared to the model group; the XLZWF-H group showed significant recovery of gastric mucosal structure, a significant reduction in inflammatory cell infiltration, and the area and density of blue collagen fibers were close to those of the control group. Figure 11 (C in the middle).

[0064] ② Serum PG I, PG II, PGR (PG I / PG II) and GAS-17 levels in each group of rats Compared with the control group, the model group showed significantly lower levels of gastrin-17 (GAS-17) and pepsinogen I (PG I), as well as a significantly lower PG I / PG II ratio (p<0.01). In contrast, the treatment groups showed significantly increased levels of GAS-17 and PG I, as well as a significantly increased PG I / PG II ratio (p<0.05, p<0.01). Figure 12 (DG in the middle).

[0065] ③ Expression levels of serum inflammatory factors (IL-1β, IL-6, TNF-α) in rats of each group Compared with the control group, the levels of IL-1β, IL-6, and TNF-α in the model group were significantly increased (p<0.01). Compared with the model group, the levels of IL-1β, IL-6, and TNF-α in the treatment group were significantly decreased (p<0.01). Figure 12 HJ in the middle).

[0066] Experimental Example 3 ① Effects of the tumor-eliminating and atrophy-treating formula on the expression of PI3K / AKT / P53 signaling pathway-related proteins and mRNAs in the gastric tissue of PL-CAG rats Compared with the control group, the expression of p-PI3K, p-Akt, and p53 proteins and mRNAs in the model group was significantly increased (p<0.01, p<0.001). Compared with the model group, the expression of L-XLZWF, M-XLZWF, H-XLZWF, and WFC, as well as p-PI3K, p-Akt / Akt, and p53 proteins and mRNAs, was significantly decreased (p<0.01, p<0.001). Figure 13 (AH in) ② Effects of the tumor-eliminating and atrophy-treating formula on the expression of autophagy-related pathway factors in the gastric tissue of PL-CAG rats Compared with the control group, the expression of Atg13, LC3, and Beclin-1 proteins and mRNAs in the model group was significantly decreased (all p < 0.0001). Compared with the model group, the expression of L-XLZWF, M-XLZWF, H-XLZWF, and WFC, as well as Atg13, LC3, and Beclin-1 proteins and mRNAs, was significantly increased (p < 0.01, p < 0.001). Figure 14 (JO in) Experiment Example 4 ① XLZWF-containing serum significantly improved cell viability and effectively reversed intestinal metaplasia-related phenotypes. The research group previously used the CCK-8 assay to detect the effect of different MNNG concentrations on the proliferation rate of GES-1 cells, and finally selected a 40 μM MNNG concentration for subsequent modeling. Figure 15 In A), treatment of GSE-1 cells with 40 μM MNNG successfully induced an intestinal metaplasia cell model (PLGC cells), characterized by a significant upregulation of the protein and mRNA expression of intestinal metaplasia markers VILLIN, KLF4, and MUC2. Figure 15 BE in Figure 16 The FH in the cell model). Subsequently, intervention with 10% serum containing the anti-tumor and anti-atrophy formula significantly improved the viability of PLGC cells (FH). Figure 16 The study found that it could effectively reverse intestinal metaplasia-related phenotypes and significantly downregulate the expression of the aforementioned intestinal metaplasia markers. Figure 17 (AG in the middle).

[0067] ② Effects of serum containing the tumor-eliminating and atrophy-treating formula on the expression of PI3K / AKT / p53 signaling pathway-related proteins and mRNAs and autophagy-related factors in PLGC cells. Compared with the control group, the expression of p-PI3K, p-Akt, mTOR, and p53 proteins and mRNA was significantly increased in the model group (p<0.01, p<0.001). Compared with the model group, the expression of p-PI3K, p-Akt / Akt, mTOR, and p53 proteins and mRNA was significantly decreased in the serum group containing the Xiaoliu Zhiwei formula (p<0.01, p<0.001). Figure 18 HO in Figure 19 (PR in the text).

[0068] Compared with the control group, the expression of Beclin-1 and LC3 I / II proteins and mRNA was significantly increased in the model group (p<0.001). Compared with the model group, the expression of Beclin-1 and LC3 I / II proteins and mRNA was significantly increased in the serum containing the anti-tumor and anti-atrophy formula (p<0.05, p<0.01). Figure 19 (SV in the text).

[0069] Experimental Example 5: Effects of the Tumor-Eliminating and Atrophy-Treating Formula on the Senescence-Related Secretion Phenotypes (VEGFR1, VEGFR2) of the Gastric Mucosa in CAG Rats In the previous study, the research group used a multi-factor composite modeling method to establish a CAG rat model. After successful modeling, the rats were randomly divided into five groups: the model group, the low-dose group of Xiaoliu Zhiwei Fang (L-XLZWF), the medium-dose group of Xiaoliu Zhiwei Fang (M-XLZWF), the high-dose group of Xiaoliu Zhiwei Fang (H-XLZWF), and the Weifuchun group (WFC). After 12 weeks of continuous administration, rat gastric tissue specimens were collected (using the same modeling and administration method as in Experiment 2). Western blot was used to detect the protein expression levels of VEGFR1, VEGFR2, and COX-2 in rat gastric tissue, and RT-qPCR was used to detect the mRNA expression levels of VEGFR1, VEGFR2, and COX-2 in rat gastric tissue. The results are as follows: Compared with the control group, the expression levels of VEGFR1, VEGFR2, and COX-2 proteins were increased in the model group (p < 0.05); compared with the model group, the expression levels of VEGFR1, VEGFR2, and COX-2 proteins were decreased in each treatment group (p < 0.05). Figure 20 (AD in the text).

[0070] Compared with the control group, the mRNA expression levels of VEGFR1, VEGFR2, and COX-2 in the model group were all increased (p < 0.05); compared with the model group, the mRNA expression levels of VEGFR1, VEGFR2, and COX-2 in each treatment group were all decreased (p < 0.05). Figure 20 (EG in the middle).

[0071] In in-vivo experiments, the Xiaoliu Zhiwei formula can effectively improve the pathological state of the gastric mucosa in CAG rats, reduce serum inflammatory indicators, inhibit SASP, and preliminarily confirm that the Xiaoliu Zhiwei formula may play a therapeutic role by inhibiting the PI3K / AKT / P53 pathway to activate autophagy, synergistically anti-inflammatory, anti-aging, and improve gastric mucosal function.

[0072] In in-vitro experiments, the serum containing the Xiaoliu Zhiwei formula can enhance the cell viability of the PLGC model and reverse the abnormal high expression of IM markers. The mechanism may be to activate autophagy by inhibiting the PI3K / AKT / P53 pathway, thereby playing a role in reversing IM.

[0073] Experimental Example 6 Observe the clinical advantages of the Xiaoliu Zhiwei formula A single-center, randomized, open, controlled clinical design was adopted. A total of 80 clinical CAG patients were collected and randomly divided into 2 groups: the control group used Weifuchun as the positive control drug, and the experimental group used the Xiaoliu Zhiwei formula. All patients received 12 weeks of intervention treatment, and the improvement degrees of clinical symptoms, gastroscopic manifestations, and pathological histology before and after treatment were observed and compared between the two groups.

[0074] Finally, 37 patients in the experimental group and 35 patients in the control group were included in the clinical controlled study, and the baseline data of the two groups were balanced and comparable. The results showed that the experimental group had significantly better curative effects than the control group in terms of improving traditional Chinese medicine syndromes, relieving core symptoms, and repairing gastric mucosal damage. Its total clinical effective rate (81.08%) and the effective rate of traditional Chinese medicine syndromes (83.78%) were both significantly higher than those of the control group, as shown in Table 2-6: Table 2 Changes in the total scores of traditional Chinese medicine syndromes before and after medication in the two groups [M(P 25 , P 75 ) or ) Note: Compared with the control group, ns P>0.05, * P<0.05, ** P<0.01, *** P<0.001; compared within the same group, # P<0.05, ## P<0.01, P<0.001. (The same as the following table of research results) Table 3 Changes in gastroscopic scores before and after medication in the two groups [M(P 25 , P 75 ) Table 4 Changes in pathological scores before and after medication in the two groups [M(P 25 , P 75 ) or ) Table 5 Comparison of clinical curative effects between the two groups (%) Table 6. Changes in AST before and after treatment [U / L] or M (P 25 P 75 )]

Claims

1. A composition for treating chronic gastritis, characterized in that, The composition, by weight, is prepared from the following raw materials: 8-12 parts of processed Pinellia ternata, 8-12 parts of Scutellaria baicalensis, 3-7 parts of Coptis chinensis, 4-8 parts of dried ginger, 12-18 parts of Pseudostellaria heterophylla, 12-18 parts of Hedyotis diffusa, 12-16 parts of Scutellaria barbata, 8-12 parts of Curcuma zedoaria, 12-20 parts of stir-fried Citrus aurantium, 18-22 parts of stir-fried germinated barley, 18-22 parts of stir-fried malt, 8-12 parts of Curcuma longa, 1-5 parts of Panax notoginseng, 8-12 parts of jujube, and 3-8 parts of prepared licorice root.

2. The composition according to claim 1, characterized in that, The composition, by weight, is prepared from the following raw materials: 10 parts of Pinellia ternata, 10 parts of Scutellaria baicalensis, 5 parts of Coptis chinensis, 6 parts of dried ginger, 15 parts of Codonopsis pilosula, 15 parts of Hedyotis diffusa, 15 parts of Scutellaria barbata, 10 parts of Curcuma zedoaria, 15 parts of stir-fried Citrus aurantium, 20 parts of stir-fried germinated barley, 20 parts of stir-fried malt, 10 parts of Curcuma longa, 3 parts of Panax notoginseng, 10 parts of jujube, and 6 parts of prepared licorice root.

3. The composition according to any one of claims 1-2, characterized in that, The composition is a pharmaceutical preparation, which is selected from: decoction, tablet, capsule, lozenge, granule, pill, powder, ointment, pill, suspension, solution, injection, suppository, ointment, spray, drop, drop pill or patch.

4. The composition according to any one of claims 1-2, characterized in that, The composition is prepared into a paste by weight parts and taken twice a day, half an hour after breakfast and dinner, one packet each time, dissolved in warm water.

5. The composition according to any one of claims 1-2, characterized in that, The main active ingredients of the composition include: isomethicone, cimicifugoside, 4-hydroxybenzoic acid, vanillin, and H1,2,4-phenylpyrogallol.

6. Use of the composition according to any one of claims 1-2 in the preparation of a medicament for treating chronic gastritis.

7. Use of the composition according to any one of claims 1-2 in the preparation of a medicament for treating chronic atrophic gastritis.

8. The use of the composition according to any one of claims 1-2 in regulating cellular homeostasis through multi-pathway synergistic regulation.

9. The application according to claim 8, characterized in that, The aforementioned multi-pathway synergy includes the PI3K / AKT / P53 signaling pathway and autophagy-related pathways.

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