Use of cyanidin-3-o-galactoside in preparation of a medicine for inhibiting pulmonary fibrosis by regulating ppbp / ccl3 / slc2a3 axis
Transcriptomic screening of the Ppbp/Ccl3/Slc2a3 axis revealed that cyanidin-3-O-galactoside regulates this axis to inhibit pulmonary fibrosis, solving the problems of adverse drug reactions and diagnostic difficulties associated with existing drugs, and achieving effective treatment and early screening for pulmonary fibrosis.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- LIAONING UNIVERSITY
- Filing Date
- 2026-04-08
- Publication Date
- 2026-06-02
AI Technical Summary
Existing pulmonary fibrosis treatments such as pirfenidone and nintedanib have adverse reactions and are expensive. Traditional diagnostic methods are costly and prone to misdiagnosis. There is a lack of effective biomarkers for early screening and personalized treatment. Existing anthocyanin research has not fully elucidated its potential regulatory pathways.
The Ppbp/Ccl3/Slc2a3 axis was screened using transcriptomics technology. Cyanidin-3-O-galactoside was used to regulate this axis to inhibit pulmonary fibrosis. Animal experiments were conducted to verify its effects in reducing the pulmonary fibrosis coefficient, malondialdehyde content, and hydroxyproline content.
It effectively alleviates pulmonary fibrosis, reduces oxidative damage to lung tissue, decreases collagen deposition, and provides the possibility of early screening and personalized treatment, avoiding the adverse reactions and high costs of traditional drugs.
Smart Images
Figure CN122124078A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biomedicine and functional food technology, specifically relating to the application of cyanidin-3-O-galactoside in the preparation of drugs that inhibit pulmonary fibrosis by regulating the Ppbp / Ccl3 / Slc2a3 axis. Background Technology
[0002] Pulmonary fibrosis (PF) is a fatal fibrotic lung disease characterized by persistent interstitial inflammation. The irreversible progression of pulmonary fibrosis results in a low median survival of only 2-3 years, with less than 20% of patients surviving for 5 years. Due to the complexity of its pathogenesis, despite a long history of research and well-documented scientific literature, only two drugs are currently available for treating PF: pirfenidone and nintedanib. However, long-term use of pirfenidone and nintedanib can cause numerous adverse reactions, such as decreased appetite and impaired lung function, and these drugs are also very expensive, placing a significant financial burden on patients in the long run.
[0003] Traditional diagnosis of pulmonary fibrosis relies on high-resolution computed tomography (HRCT) and lung biopsy, methods that are costly and risky, and struggle to reveal the underlying molecular causes and progression mechanisms of the disease. Furthermore, pulmonary fibrosis is easily confused with other lung diseases, leading to misdiagnosis and delayed treatment. Against this backdrop, identifying reliable biomarkers that reflect the disease's origin, progression, and treatment response is crucial for optimizing diagnostic processes, enabling early screening, and personalized treatment. Transcriptomics sequencing technology provides a powerful tool for systematically analyzing disease-related gene expression profiles and uncovering potential regulatory molecules, offering technical support for the development of precision medicine for pulmonary fibrosis.
[0004] Anthocyanins, a class of polyphenolic compounds widely found in natural plants, have gradually attracted attention due to their significant anti-inflammatory, antioxidant, and anti-fibrotic activities. Among them, cyanidin-3-O-galactoside (C3G), as a representative anthocyanin with abundant content and well-defined activity, has been shown in previous studies to reduce the transformation of lung fibroblasts into myofibroblasts and collagen synthesis by inhibiting the transforming growth factor-β1 (TGF-β1) / Smad signaling pathway. Simultaneously, it scavenge reactive oxygen species, inhibit oxidative stress, and alleviate inflammatory damage and fibrotic deposition in lung tissue. However, current research on C3G largely focuses on the regulation of classical pathways, leaving many research gaps and numerous potential regulatory pathways and targets yet to be further elucidated. This invention aims to alleviate pulmonary fibrosis by using C3G to regulate the Ppbp / Ccl3 / Slc2a3 axis screened using transcriptomics technology. Summary of the Invention
[0005] This invention aims to find new regulatory axes to alleviate pulmonary fibrosis. It proposes using transcriptomics technology to screen the Ppbp / Ccl3 / Slc2a3 axis and to apply C3G regulation of this axis to alleviate pulmonary fibrosis. A bleomycin-induced pulmonary fibrosis mouse model was used to verify the effect of cyanidin-3-O-galactoside regulating the Ppbp / Ccl3 / Slc2a3 axis in alleviating pulmonary fibrosis. The effects on inflammation accumulation, oxidative stress, and tissue damage in the pulmonary fibrosis model mice were evaluated to verify the inhibitory effect of cyanidin-3-O-galactoside on pulmonary fibrosis through the Ppbp / Ccl3 / Slc2a3 axis.
[0006] The technical solution adopted in this invention is:
[0007] Application of cyanidin-3-O-galactoside in the preparation of drugs that inhibit pulmonary fibrosis by regulating the Ppbp / Ccl3 / Slc2a3 axis.
[0008] Furthermore, in the above-mentioned application, cyanidin-3-O-galactoside inhibits the increase in lung coefficient caused by pulmonary fibrosis by regulating the Ppbp / Ccl3 / Slc2a3 signaling axis.
[0009] Furthermore, in the above-mentioned application, cyanidin-3-O-galactoside inhibits the increase in malondialdehyde content caused by pulmonary fibrosis by regulating the Ppbp / Ccl3 / Slc2a3 signaling axis.
[0010] Furthermore, in the above-mentioned application, cyanidin-3-O-galactoside inhibits the increase in hydroxyproline caused by pulmonary fibrosis by regulating the Ppbp / Ccl3 / Slc2a3 signaling axis.
[0011] Furthermore, in the above-mentioned application, cyanidin-3-O-galactoside alleviates fibrotic lesions by regulating the Ppbp / Ccl3 / Slc2a3 signaling axis.
[0012] Furthermore, in the above application, the dosage of cyanidin-3-O-galactoside in animals is 400 mg / kg body weight.
[0013] Furthermore, in the above applications, the Ppbp / Ccl3 / Slc2a3 signaling axis is obtained through transcriptomics screening.
[0014] Furthermore, in the above-mentioned applications, the cyanidin-3-O-galactoside was obtained through network pharmacology screening.
[0015] Furthermore, in the above application, the core target of pulmonary fibrosis is Ccl3.
[0016] Furthermore, in the above application, the core target of pulmonary fibrosis, Ccl3, was obtained through transcriptomic screening.
[0017] The beneficial effects of this invention are as follows: Transcriptome sequencing data analysis revealed the core target of pulmonary fibrosis, Ccl3, and its regulatory network Ppbp / Ccl3 / Slc2a3. Network pharmacology analysis identified cyanidin-3-O-galactoside as the most relevant anthocyanin for pulmonary fibrosis. Animal experiments verified that cyanidin-3-O-galactoside can reduce the pulmonary fibrosis coefficient, decrease malondialdehyde content, and reduce hydroxyproline content. Therefore, it is demonstrated that cyanidin-3-O-galactoside can alleviate pulmonary fibrosis through Ppbp / Ccl3 / Slc2a3 signal regulation. Attached Figure Description
[0018] Figure 1 This is a Venn diagram of the core targets of pulmonary fibrosis analyzed by transcriptomics.
[0019] Figure 2 This is a Venn diagram showing the targets of anthocyanin-related active ingredients associated with pulmonary fibrosis. A: Core targets related to PF; B: Core targets of cyanidin-related anthocyanins; C: Core targets of delphinidin-related anthocyanins; D: Core targets of mallow-related anthocyanins; E: Core targets of pelargonidin-related anthocyanins; F: Core targets of peony-related anthocyanins; G: Core targets of morning glory-related anthocyanins.
[0020] Figure 3 The graph shows the comparison results of lung coefficients of mice in each group. The comparison between WT-Model and WT-Control groups, and between WT-Model and WT-Model-C3G groups is shown. *p<0.05, **p<0.01, ***p<0.001.
[0021] Figure 4 This is a graph showing the comparison of malondialdehyde (MDA) content in the lung tissue of mice in each group. Comparison between WT-Model and WT-Control groups, and between WT-Model and WT-Model-C3G groups. *p<0.05, **p<0.01, ***p<0.001.
[0022] Figure 5 This is a graph showing the comparison of hydroxyproline (HYP) content in the lung tissue of mice in each group. Comparison between WT-Model and WT-Control groups, and between WT-Model and WT-Model-C3G groups. *p<0.05, **p<0.01, ***p<0.001.
[0023] Figure 6 This is a comparison chart of HE, Masson, and Sirius red staining of lung tissue from different groups of mice. Detailed Implementation
[0024] Example 1
[0025] (I) Drug Preparation
[0026] 10% Chloral Hydrate: Weigh 10.0g of chloral hydrate powder, add 100mL of double-distilled water, dissolve and filter, and store away from light.
[0027] Preparation of Bleomycin (BLM) solution: Dissolve 25 mg BLM in 750 μL of physiological saline to prepare a 50 U / mL stock solution, dispensed into 150 μL vials. When using, add 7.35 mL of physiological saline to each 150 μL vial to dilute to a 1 U / mL working solution.
[0028] (ii) Sequencing tissue acquisition
[0029] Experimental animal grouping: 20 male C57BL / 6 mice and 20 male Ccl3 gene-deficient C57BL / 6 mice, weighing 20±2g, were randomly divided into two groups of 10 each, as follows:
[0030] (1) Normal control group (WT-blank);
[0031] (2) Pulmonary fibrosis model group (WT-pulmonary fibrosis model);
[0032] (3) KO-Ccl3 control group (KO-Ccl3 blank);
[0033] (4) KO-Ccl3 pulmonary fibrosis model group (KO-Ccl3 pulmonary fibrosis model).
[0034] Healthy male mice aged 6-8 weeks were selected and acclimatized to their environment for one week before model initiation. Except for two control groups, the remaining mice underwent tracheal infusion of 0.1 mL BLM (1 U / mL) for model initiation. All four groups of mice were administered physiological saline for 7 consecutive days. After 7 days, lung tissue samples were taken from the same portion of each group (n=5 per group) for sequencing.
[0035] (III) Ppbp / Ccl3 / Slc2a3 signals
[0036] Transcriptome sequencing data from WT-blank group and WT-pulmonary fibrosis model group mice were analyzed, such as... Figure 1 As shown, the core target of pulmonary fibrosis, Ccl3, was screened using differentially expressed genes in pulmonary fibrosis, PPI, PredGenes, and WGCNA. The interaction analysis of the WT-blank group, WT-pulmonary fibrosis model group, KO-Ccl3 blank group, and KO-Ccl3 pulmonary fibrosis model group yielded the Ppbp / Ccl3 / Slc2a3 signal axis.
[0037] (iv) Anthocyanin screening
[0038] like Figure 2 As shown, the molecular structures of delphinidin, malvidin, cyanidin, morning glory pigment, peony pigment, and geranium pigment were retrieved using Pubchem and imported into three databases for small molecule compound target gene prediction: SwisstargetPrediction, SEAPrediction, and Superhed. Based on the English name for pulmonary fibrosis, the GeneCard online database was searched for the name of pulmonary fibrosis. Anthocyanins and target genes for pulmonary fibrosis were screened, and cyanidin-3-O-galactoside, which is most associated with pulmonary fibrosis, was identified.
[0039] Example 2
[0040] (I) Drug Preparation
[0041] Cyanide-3-O-galactoside: Purchase monomer from Hebei Wangyou Biotechnology, purity 95-98%.
[0042] The preparation method of the 10% chloral hydrate and bleomycin solution is the same as in Example 1.
[0043] (ii) Efficacy test
[0044] Animal husbandry and handling: Laboratory animals were housed for one week to acclimatize to the laboratory conditions, then separated into individual cages. The room was kept under sterile conditions, and bedding was changed every two days. The laboratory lighting was controlled, with 12 hours of light followed by 12 hours of darkness. The room temperature was maintained at 20±1℃, and the humidity at 55±10%.
[0045] Experimental animal grouping: 30 male C57BL / 6 mice, weighing 20±2g, were randomly divided into 3 groups as follows, with 10 mice in each group.
[0046] (1) Normal control group (WT-Control);
[0047] (2) Pulmonary fibrosis model group (WT-Model);
[0048] (3) Drug administration group: Cyanidin-3-O-galactoside group 400 mg / kg (WT-Model-C3G);
[0049] Healthy male mice aged 6-8 weeks were selected and acclimatized to their environment for one week before modeling was initiated. Except for the normal control group, the remaining mice underwent endotracheal infusion of 0.1 mL BLM (1 U / mL) for modeling. Simultaneously with modeling, medication was administered. While the normal control group and the pulmonary fibrosis model group received saline solution, the medication groups received cyanidin-3-O-galactoside 400 mg / kg. The mice's condition was observed daily, mortality was recorded, and body weight was recorded every other day. Results were assessed after 5 weeks of experimentation.
[0050] (III) Test Results
[0051] 1. After blood collection, mice were sacrificed, lung tissue was collected and weighed, and the lung coefficient was calculated. The comparison results of the lung coefficients are shown below. Figure 3 .
[0052] like Figure 3 As shown, the lung coefficient of mice in the pulmonary fibrosis model group was significantly higher than that in the normal group (p<0.001). Cyanidin-3-O-galactoside administration could intervene in the increase in lung coefficient caused by pulmonary fibrosis. Compared with the pulmonary fibrosis model group, the administration group could significantly inhibit the increase in lung coefficient in mice (p<0.05). The experimental results indicate that cyanidin-3-O-galactoside has a significant dose-dependent intervention effect on the increase in lung coefficient during pulmonary fibrosis in mice.
[0053] 2. Detect the malondialdehyde (MDA) content in lung tissue. Results are shown below. Figure 4 .
[0054] like Figure 4 As shown, the malondialdehyde (MDA) content in the pulmonary fibrosis model group was significantly higher than that in the control group (p<0.001), and the cyanidin-3-O-galactoside administration group could intervene in the increase in MDA content caused by pulmonary fibrosis. The results indicate that cyanidin-3-O-galactoside has a good ability to alleviate oxidative damage in lung tissue, and its intervention effect on pulmonary fibrosis is significant and dose-dependent.
[0055] 3. The content of hydroxyproline, a marker of fibrosis, in lung tissue was detected. Results are shown below. Figure 5 .
[0056] like Figure 5 As shown, the hydroxyproline (HYP) content in the pulmonary fibrosis model mice was significantly higher than that in the normal group (p<0.001). Cyanidin-3-O-galactoside administration could intervene in the increase of hydroxyproline induced by pulmonary fibrosis. Compared with the pulmonary fibrosis model group, the administration group significantly inhibited the increase of hydroxyproline in mice (p<0.05). These results indicate that cyanidin-3-O-galactoside can reduce the increase of hydroxyproline in lung tissue, reduce excessive deposition of extracellular matrix, and thus alleviate bleomycin-induced pulmonary fibrosis.
[0057] 4. HE, Masson's red, and Sirius red staining were used to observe the structure and morphology of lung tissue in mice with pulmonary fibrosis. The results are shown in [Figure number missing]. Figure 6 .
[0058] like Figure 6 As shown, HE, Masson's red, and Sirius red (SR) section results indicated that the lung tissue structure of mice in the blank control group was normal, with no proliferative fibrosis. In the pulmonary fibrosis model group, the lung tissue of mice showed severe inflammation and damage, with extensive collagen deposition and complete destruction of alveolar structure. Compared with the model group, the lung tissue damage in the cyanidin-3-O-galactoside group was significantly improved, with alveolar tissue maintaining integrity and minimal inflammatory infiltration. The high-dose group showed a more significant effect. These results demonstrate that cyanidin-3-O-galactoside can alleviate bleomycin-induced pulmonary fibrosis.
Claims
1. Application of cyanidin-3-O-galactoside in the preparation of drugs that inhibit pulmonary fibrosis by regulating the Ppbp / Ccl3 / Slc2a3 axis.
2. The application according to claim 1, characterized in that, The cyanidin-3-O-galactoside inhibits the increase in lung coefficient caused by pulmonary fibrosis by regulating the Ppbp / Ccl3 / Slc2a3 signaling axis.
3. The application according to claim 1, characterized in that, The cyanidin-3-O-galactoside inhibits the increase in malondialdehyde content caused by pulmonary fibrosis by regulating the Ppbp / Ccl3 / Slc2a3 signaling axis.
4. The application according to claim 1, characterized in that, The cyanidin-3-O-galactoside inhibits the increase in hydroxyproline caused by pulmonary fibrosis by regulating the Ppbp / Ccl3 / Slc2a3 signaling axis.
5. The application according to claim 1, characterized in that, The cyanidin-3-O-galactoside alleviates fibrotic lesions by regulating the Ppbp / Ccl3 / Slc2a3 signaling axis.
6. The application according to claim 1, characterized in that, The dosage of cyanidin-3-O-galactoside in animals is 400 mg / kg body weight.
7. The application according to claim 1, characterized in that, The Ppbp / Ccl3 / Slc2a3 signaling axis was obtained through transcriptomics screening.
8. The application according to claim 1, characterized in that, The cyanidin-3-O-galactoside was obtained through network pharmacology screening.
9. The application according to claim 1, characterized in that, The core target of pulmonary fibrosis is Ccl3.
10. The application according to claim 9, characterized in that, The core target of pulmonary fibrosis, Ccl3, was obtained through transcriptomics screening.