Application of party ginseng glycosides in preparation of drugs for preventing or treating sepsis

Abstract

The application discloses application of atractylenolide in preparation of a medicine for preventing or treating sepsis, and relates to the technical field of biological medicine, and has the technical points that: the application carries out experimental research on atractylenolide in prevention and / or treatment of sepsis by constructing an LPS-induced acute sepsis model, and the results show that atractylenolide has a good protective effect on LPS-induced sepsis, and has small side effects, and can be used for preparing a protective medicine for treating or preventing sepsis. The application provides a new use of atractylenolide in preparation of a medicine for treating sepsis, and has remarkable effects in improving tissue damage and playing an anti-inflammatory role, and has small side effects.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to the use of codonopsis pilosula in preparing a drug for preventing or treating sepsis. Background Art

[0002] Sepsis is a common pathological process in many infectious and non-infectious diseases, such as burns, trauma, and pancreatitis. If left untreated, it can lead to multiple organ dysfunction and ultimately death. Sepsis is the leading cause of death in intensive care units (ICUs) worldwide, with a mortality rate exceeding that of acute myocardial infarction and ranking first among non-tumor diseases. Early detection and timely initiation of treatment are crucial for achieving a good outcome in patients with sepsis. Sepsis is typically characterized by a systemic immune-inflammatory response, with common symptoms including fever, tachycardia, tachypnea and respiratory changes, and abnormal psychological status. Current treatment options for sepsis still rely on combination antibiotic therapy and multi-organ support therapy. However, with the prevalence of drug-resistant strains, existing clinical treatments are gradually falling into a predicament, and there is an urgent need to develop new and effective methods for treating sepsis.

[0003] Numerous studies have shown that immune dysregulation and systemic inflammatory responses in patients with sepsis are significant contributors to mortality. Therefore, effectively alleviating or reversing this immune-inflammatory imbalance may be key to treating sepsis. Macrophages play a crucial role in the innate immune response. They recognize and phagocytose pathogens through pathogen-associated molecular pattern recognition receptors, including toll-like receptors (TLRs), oligonucleotide-binding domain-like receptors (NLRs), and retinoic acid-induced gene-like receptors (RLRs), and secrete inflammatory cytokines such as IL-1, IL-2, IL-6, IFN-γ, and TNF-α to combat infection. Macrophages in patients with sepsis become abnormally active, leading to excessive release of inflammatory cytokines, resulting in a persistent inflammatory response and exacerbated tissue damage. Therefore, early inhibition of macrophage secretion or production of inflammatory cytokines may effectively alleviate sepsis.

[0004] For centuries, Codonopsis pilosula, a perennial herb in the Campanulaceae family, has been widely used in East and Southeast Asia as both a food and medicine, tonifying Qi and blood, enhancing immunity, improving appetite, and delaying aging. Lobetyolin (LBT), a bioactive compound extracted from Codonopsis pilosula, has been shown to possess diverse biological and pharmacological properties, including cardioprotective, anti-inflammatory, antioxidant, and anti-tumor properties. Despite the literature reporting the multiple pharmacological activities of lobetyolin, its role in the prevention and treatment of sepsis has been unknown. Summary of the Invention

[0005] The present invention aims to solve the above problems. The present invention discovers the clinical application value of codonopsis pilosula in the prevention and treatment of sepsis, and proposes that codonopsis pilosula has a good protective effect on sepsis, improves pathological damage to the lungs and livers of mice, and has significant anti-inflammatory effects with minimal side effects.

[0006] In order to achieve the above object, the technical solution of the present invention is as follows: the molecular formula of the dangshen glycoside is C 20 H 28 O8, molecular weight 396.43, CAS No.: 129277-38-9, structural formula

[0007]

[0008] As shown below:

[0009] The present invention provides the use of codonopsis pilosula in preparing a medicine for preventing and / or treating sepsis.

[0010] Preferably, the application includes:

[0011] (A1) preparing a product for improving the survival rate of sepsis patients;

[0012] (A2) preparing a product for reducing the expression level of inflammatory factors in the serum of sepsis patients;

[0013] (A3) preparing a product for alleviating lung and liver damage in patients with sepsis;

[0014] (A4) preparing a product for restoring the regulation of inflammatory factors in peritoneal macrophages in patients with sepsis;

[0015] (A5) Preparing a product for preventing sepsis.

[0016] Preferably, codonopsis pilosula alleviates sepsis by reducing inflammatory response.

[0017] Preferably, the reduction of inflammatory response includes down-regulating the expression of proinflammatory factors and antibacterial response genes; the proinflammatory factors include TNF-α, IL-6 and IL-1β, and the antibacterial response genes include Cxcl10, Tgtp1, and Gbp5.

[0018] Preferably, the drug comprises codonopsis pilosula and pharmaceutically acceptable excipients.

[0019] Preferably, the pharmaceutically acceptable excipients include any one or more of fillers, disintegrants, binders, lubricants and flavoring agents.

[0020] Compared with the existing technology, this solution has the following beneficial effects:

[0021] The present invention discloses the use of codonopsis glycoside in the preparation of a protective drug against LPS-induced sepsis. Animal and cell experiments were conducted, respectively. In C57 wild-type mice, a sepsis model was simulated by intraperitoneal injection of LPS, and codonopsis glycoside was administered intraperitoneally to demonstrate its application in sepsis protection. Results showed that compared with the untreated model group, the codonopsis glycoside-treated model significantly improved pathological damage in the mouse lung and liver tissues, reduced the levels of inflammatory factors in peripheral blood, and reduced the responses of inflammatory factors and antimicrobial signals in peritoneal macrophages. Cell experiments used primary C57 mouse macrophages as experimental subjects, and LPS-stimulated in vitro sepsis cell models were constructed to investigate the effect of codonopsis glycoside on the macrophage response to LPS stimulation. Results showed that compared with the untreated LPS-only group, the LPS-treated group significantly downregulated the levels of proinflammatory factors in the cell culture supernatant and cells. Therefore, codonopsis glycoside can play a role in preventing and treating sepsis and can be used to prepare drugs for the treatment of sepsis, with good efficacy and low toxic side effects, and has considerable potential value.

[0022] The present invention provides more possible candidate drugs for treating sepsis.

[0023] The present invention provides a new use of codonopsis pilosula in treating sepsis, which has significant effects in improving tissue damage and exerting anti-inflammatory effects with few side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 Schematic diagram of the protective effect of dangshen glycoside pretreatment on sepsis in mice according to an embodiment of the present invention;

[0025] Figure 2 Schematic diagram of the results of related gene detection of mouse peritoneal macrophages in an embodiment of the present invention;

[0026] Figure 3 2 is a schematic diagram of GO analysis of the gene differential expression analysis of RNA sequencing results of mouse peritoneal macrophages after treatment with codonopsis pilosula in an embodiment of the present invention;

[0027] Figure 4 Schematic diagram of KEGG analysis of gene differential expression of RNA sequencing results of mouse peritoneal macrophages after treatment with codonopsis pilosula in an embodiment of the present invention;

[0028] Figure 5 Schematic diagram of RNA-seq analysis of mouse peritoneal macrophages pretreated with codonopsis pilosula and then stimulated with LPS in an embodiment of the present invention;

[0029] Figure 6 This is a schematic diagram of the inhibition of TNF-α and IL-6 expression in macrophages by codonopsis pilosula in an embodiment of the present invention. DETAILED DESCRIPTION

[0030] In order to enable those skilled in the art to better understand the present invention, the technical solution of the present invention will be further described in detail below in conjunction with the embodiments of the present invention and the accompanying drawings. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts should fall within the scope of protection of the present invention.

[0031] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0032] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the present invention. The present description and examples are intended to be illustrative only.

[0033] The terms "include," "comprising," "having," and "containing" used herein are open-ended terms, meaning including but not limited to, the present invention. It should be noted that the embodiments and features of the embodiments of the present invention may be combined with each other unless there is a conflict. The present invention will be described in detail below with reference to the embodiments.

[0034] Example:

[0035] The present invention studies the use of codonopsis glycoside in the preparation of LPS-induced sepsis protective drugs through animal and cell experiments. The animals are C57 wild-type mice as experimental subjects. The sepsis model is simulated by intraperitoneal injection of LPS, and codonopsis glycoside is administered by intraperitoneal injection to explore its application in sepsis protection. The cell experiment uses mouse peritoneal macrophages as experimental subjects. The cells are stimulated with LPS to construct an in vitro sepsis cell model to explore the effect of codonopsis glycoside on macrophages.

[0036] Animal Husbandry: Male C57BL / 6 mice (8 weeks old) were obtained from the Model Animal Research Center of Nanjing University. Experiments were conducted after a 1-2 week acclimation period. Four to six mice were housed in a cage and provided with regularly replaced sterilized standard mouse feed and room-temperature drinking water. The animals were divided into two groups: the LPS model group (LPS) and the codonopsis pilosula group (LPS + LBT).

[0037] Animal modeling and drug administration: Mice were first intraperitoneally injected with 10mM LBT or phosphate-buffered saline (PBS). 24 hours later, lipopolysaccharide (LPS) (5mg / kg) was intraperitoneally injected. Peripheral blood samples, lung tissue, and liver tissue were collected 4 hours later. Successful establishment of an LPS-induced sepsis mouse model was confirmed by observing the survival of the mice and subsequently measuring pathological damage in their lungs and liver tissues, as well as pro-inflammatory cytokine levels in peripheral blood and peritoneal macrophages.

[0038] Experimental Conclusion

[0039] refer to Figure 1 As shown in the figure, the protective effect of LBT pretreatment on sepsis in mice. C57BL / 6 mice were intraperitoneally injected with LBT (10 mM) or PBS, and 24 hours later, LPS (5 mg / kg) was injected. Figure 1 A is the analysis of mouse survival rate; Figure 1 B- Figure 1 D is ELISA to detect IL-6 ( Figure 1 B), TNF-α ( Figure 1 C), IL-1β ( Figure 1 D) secretion level; Figure 1 E and Figure 1 F is H&E staining of mouse lung and liver tissues.

[0040] refer to Figure 2 Figure 2 shows the results of related gene detection in mouse peritoneal macrophages. Mouse peritoneal macrophages were treated with 10 μM LBT or PBS for 24 hours and then RNA was extracted and subjected to RNA sequencing. Figure 2 A is the difference in gene changes between the 10 μM codonopsis pilosula treatment group and the control group; Figure 2 B is a volcano plot showing the specific changes of all genes after RNA sequencing; Figure 2 C is a Venn diagram showing differentially regulated genes after treatment with leucoside (LBT) (10 μM) or PBS; Figure 2 D and Figure 2 E is the top 10 upregulated proteins ranked by FPKM fragment expression value ( Figure 2 D) and downregulation ( Figure 2 E) Differential genes.

[0041] refer to Figure 3 As shown, GO analysis of the gene differential expression analysis of RNA sequencing results of mouse peritoneal macrophages after treatment with 10 μM of LBT for 24 hours. Figure 3 A is the GO grouping of the 20 most significant changes in differentially expressed genes. Figure 3 B- Figure 3C is the volcano plot of the difference in bacterial-related genes in mouse macrophages after LBT treatment ( Figure 3 B), and the top 10 significantly changed genes with FPKM values ​​> 0 ( Figure 3 C). Figure 3 D- Figure 3 E is the volcano plot of the down-regulated genes in mouse macrophages in response to lipopolysaccharide after LBT treatment ( Figure 3 D), and the top 10 significantly changed genes with FPKM values ​​> 0 ( Figure 3 E).

[0042] refer to Figure 4 As shown, KEGG analysis of gene differential expression analysis of RNA sequencing results of mouse peritoneal macrophages after treatment with 10 μM of LBT for 24 hours. Figure 4 A is a scatter plot of the top 20 most significant KEGG pathways in mouse peritoneal macrophages after LBT treatment compared with the control group. Volcano plot of gene differences in cytokine-cytokine receptor interactions in macrophages after LBT treatment ( Figure 4 B), and the top 10 significantly downregulated genes with FPKM values ​​> 0 ( Figure 4 C) Volcano plot of gene differences in macrophages responding to Toll ligand signaling pathway after LBT treatment ( Figure 4 D), and the top 10 significantly downregulated genes with FPKM values ​​> 0 ( Figure 4 E).

[0043] refer to Figure 5 As shown, mouse peritoneal macrophages were pretreated with 10 μM dapoxetine (LBT) and then stimulated with 100 ng / ml LPS, and the peritoneal macrophages were collected for RNA-seq analysis. Figure 5 A is the volcano plot of all genes obtained by RNA sequencing. Figure 5 B- Figure 5 C is a scatter plot showing the top 20 most significant GO enriched gene sets in macrophages after LBT and LPS treatment compared with the LPS control group ( Figure 5 B) and KEGG pathway ( Figure 5 C). Figure 5 D- Figure 5 E is the volcano plot of gene differences related to bacterial response in mouse macrophages after LBT treatment ( Figure 5 D), and the top 10 significantly changed genes with FPKM values ​​> 0 ( Figure 5 E). Figure 5 F- Figure 5 G is the volcano plot of gene differences in cytokine-cytokine receptor interactions in macrophages after LBT treatment ( Figure 5 F), and the top 10 significantly changed genes with FPKM values ​​> 0 ( Figure 5 G).

[0044] refer to Figure 6 As shown, LBT inhibited the expression of TNF-α and IL-6 in macrophages. RT-PCR ( Figure 6 A) and ELISA ( Figure 6 B) Analysis of IL-6 and TNF-α levels in peritoneal macrophages of mice after LBT pretreatment and LPS treatment at different time points. RT-PCR analysis of IL-6 and TNF-α levels in peritoneal macrophages of mice pretreated with LBT for 24 hours and then treated with poly(I:C), CPG, and PGN for another 3 hours. Figure 6 C) and TNF-α ( Figure 6 D) expression levels.

[0045] In summary, the current results indicate that codonopsis pilosula can alleviate LPS-induced sepsis in mice.

[0046] The above specific embodiments are merely explanations of the present invention and are not limitations of the present invention. After reading this specification, those skilled in the art may make non-creative modifications to the embodiments as needed. However, as long as they are within the scope of the claims of the present invention, they are protected by patent law.

Claims

1. Use of codonopsis pilosula in the preparation of drugs for preventing and / or treating sepsis.

2. The use according to claim 1, characterized in that The applications include: (A1) preparing a drug for improving the survival rate of sepsis patients; (A2) preparing a drug for reducing the expression level of inflammatory factors in the serum of sepsis patients; (A3) preparing a medicament for alleviating lung and liver damage in patients with sepsis; (A4) preparing a drug for restoring the regulation of inflammatory factors in peritoneal macrophages in patients with sepsis; (A5) Prepare a medicament for preventing sepsis.

3. The use according to claim 1, characterized in that: Codonopsis pilosula alleviates sepsis by reducing inflammatory response.

4. The use according to claim 3, characterized in that: The reduction of inflammatory response includes down-regulating the expression of pro-inflammatory factors and anti-bacterial response genes; the pro-inflammatory factors include TNF-α, IL-6 and IL-1β, and the anti-bacterial response genes include Cxcl10, Tgtp1 and Gbp5.

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