Use of macrophyllainulin in preparation of medicine for treating sepsis intestinal injury
Patent Information
- Application Number
- CN202611273627.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-08-21
- Publication Date
- 2026-09-25
AI Technical Summary
但是，大花旋覆花内酯能否用于改善脓毒症肠道屏障功能障碍及提高脓毒症生存率，目前尚无相关报道
[0017]1）显著提高脓毒症动物生存率，改善整体预后：大花旋覆花内酯可显著延长脓毒症小鼠生存期，缓解模型小鼠体重下降及结直肠缩短，有效减轻脓毒症所致肠道屏障结构破坏。
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Figure CN122805634A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biomedical technology, specifically relating to the application of Inula japonica lactone in the preparation of drugs for treating septic intestinal injury. Background Technology
[0002] Sepsis is a leading cause of disability and death among hospitalized patients (especially those in intensive care units). Its pathogenesis is complex, involving inflammatory imbalances and immune dysfunction, ultimately leading to organ failure. The intestine is the most frequently affected initiating organ in sepsis. During the course of the disease, increased intestinal barrier permeability leads to bacterial translocation and cytokine storms. Clinically, there is a lack of specific and safe drugs for targeted treatment of sepsis-related intestinal damage, and gaps exist in early prevention and late-stage treatment.
[0003] Existing research indicates that TNF receptor-associated factor 6 (TRAF6) is a key adaptor protein in TLR signaling, participating in inflammatory responses through pathways such as NF-κB and playing a central role in intestinal inflammation and intestinal mucosal damage. Britanninn, a natural active ingredient extracted from *Inula japonica*, has been validated to possess anti-inflammatory, antibacterial, and anticancer pharmacological activities, and can alleviate inflammation by inhibiting macrophage inflammasome activation. However, whether Britanninn can be used to improve intestinal barrier dysfunction and increase sepsis survival rates remains unreported. Summary of the Invention
[0004] In view of the above-mentioned shortcomings of the prior art, the main objective of this invention is to provide a new medical use for the natural product Inula japonica lactone, for the preparation of a drug for treating sepsis intestinal injury. By targeting the TRAF6 / NF-κB signaling pathway, it improves the intestinal barrier function in sepsis, providing a new candidate drug and clinical translation basis for targeted therapy of sepsis intestinal injury.
[0005] To achieve the above objectives, the present invention adopts the following technical solution:
[0006] In a first aspect, the invention provides the use of Inula japonica lactone in the preparation of a medicament for treating septic intestinal injury, wherein the molecular formula of Inula japonica lactone is C 19 H 26 O7, chemical formula is .
[0007] Preferably, the sepsis-related intestinal injury refers to sepsis-related intestinal barrier function impairment.
[0008] Preferably, the drug is a formulation prepared by using an effective dose of Inula japonica lactone as the active ingredient, plus pharmaceutically acceptable excipients.
[0009] Preferably, the drug is formulated into a compound preparation in combination with antibiotics, anti-inflammatory drugs, or enteral nutrition protectants.
[0010] Preferably, the formulation is selected from at least one of the following formulations: oral dosage form, tablet, injection, or capsule.
[0011] Preferably, the formulation is an injectable preparation, and the administration method includes intravenous injection or subcutaneous injection.
[0012] A second aspect of the present invention provides a pharmaceutical composition for treating septic intestinal injury, comprising an effective dose of inula lactone as an active ingredient, wherein the molecular formula of inula lactone is C 19 H 26 O7, chemical formula is .
[0013] A third aspect of the present invention provides the use of Inula japonica lactone in the preparation of a drug that inhibits the expression of pro-inflammatory factors IL-6, TNF-α and IL-1β, wherein the molecular formula of Inula japonica lactone is C 19 H 26 O7, chemical formula is .
[0014] A fourth aspect of the present invention provides the use of Inula japonica lactone in the preparation of inhibitors of the TRAF6-NF-κB signaling pathway, wherein the molecular formula of Inula japonica lactone is C 19 H 26 O7, chemical formula is This is achieved by inhibiting p-P65 expression and promoting ZO-1 expression.
[0015] In this invention, Inula japonica lactone is used to prepare a drug for treating intestinal injury in sepsis. Its mechanism of action is as follows: Inula japonica lactone inhibits TRAF6 by targeting and inhibiting NF-κB pathway activation (downregulating p-P65 expression), reducing the expression of pro-inflammatory factors IL-6, TNF-α and IL-1β in serum and intestinal tissue, while significantly upregulating the expression of intestinal tight junction protein ZO-1, thereby improving intestinal barrier dysfunction in sepsis and prolonging the survival of sepsis patients.
[0016] Compared with the prior art, the present invention has the following beneficial effects:
[0017] 1) Significantly improves the survival rate and overall prognosis of septic animals: Inula japonica lactone can significantly prolong the survival time of septic mice, alleviate the weight loss and colon and rectum shortening in model mice, and effectively reduce the damage to the intestinal barrier structure caused by sepsis.
[0018] 2) Effectively protects and improves intestinal barrier function: In in vitro human and mouse intestinal epithelial cell inflammation models, Inula japonica lactone can significantly reverse LPS-induced downregulation of intestinal tight junction protein ZO-1, maintain the integrity of the intestinal mucosal barrier, and improve sepsis intestinal barrier dysfunction.
[0019] 3) Strong inhibition of systemic and local inflammatory response: Inula japonica lactone can significantly reduce the expression levels of pro-inflammatory factors IL-6, TNF-α and IL-1β in serum (systemic) and colorectal tissue (local) in the body under sepsis state, reduce intestinal inflammatory response and alleviate intestinal inflammatory damage.
[0020] 4) Clear target and high drug safety: This invention has verified through experiments that Inula japonica lactone exerts its anti-septic intestinal injury effect by targeting and inhibiting the TRAF6 / NF-κB signaling pathway, and the mechanism of action is clear; Inula japonica lactone is a natural plant-derived active ingredient with good biosafety. Attached Figure Description
[0021] Figure 1 The results of this study were used to verify the effects of Britannin on the survival and intestinal phenotype of septic mice.
[0022] Figure 2 The results of Britannin's regulation of the expression of the intestinal barrier tight junction protein ZO-1 were verified in this example (in vivo and in vitro).
[0023] Figure 3 The results of this study were used to verify the inhibitory effect of Britannin on the secretion of systemic and local intestinal inflammatory factors.
[0024] Figure 4 The results of this study demonstrate the mechanism by which Britannin functions through the TRAF6 / NF-κB pathway.
[0025] Figure 5 This example demonstrates the results of Britannin's anti-inflammatory effect by inhibiting macrophage M1 polarization.
[0026] Figure 6 This example demonstrates the results of Britannin exerting its anti-inflammatory effect by promoting M2 polarization in macrophages. Detailed Implementation
[0027] To more fully understand and demonstrate the technical solutions, objectives, and advantages of the present invention, the technical effects produced by the present invention will be further described in detail and completely below with reference to the accompanying drawings and specific embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. It should be noted that other embodiments obtained by those skilled in the art without departing from the concept of the present invention are all within the protection scope of the present invention.
[0028] The Britannin (CAS No. 33627-28-0) used in the following examples has the molecular formula C 19 H 26 O7, chemical formula is And it is commercially available.
[0029] Example 1: Effects of Britannin on survival and intestinal phenotype in septic mice
[0030] A mouse model of LPS-induced sepsis was established, and mice were divided into a control group, a Britannin monotherapy group, an LPS model group, and an LPS+Britannin treatment group. Survival rate and body weight changes were recorded after drug administration; colon and rectum length were measured by dissection, and the degree of intestinal barrier damage was assessed by histological staining.
[0031] The results are as follows Figure 1 As shown, the LPS+Britannin group had significantly prolonged survival, and significant relief from weight loss and colonic shortening.
[0032] Example 2: Regulation of intestinal barrier tight junction protein ZO-1 expression by Britannin (in vivo and in vitro)
[0033] Proteins were extracted from the intestinal tissues of mice in each group, and ZO-1 expression was detected by Western blotting. Simultaneously, LPS inflammation models of mouse intestinal epithelial MODE-K cells and human intestinal epithelial NCM460 cells were established and treated with Britannin, and changes in ZO-1 levels were detected.
[0034] The results are as follows Figure 2 As shown, Britannin can significantly reverse the LPS-induced downregulation of ZO-1 protein expression.
[0035] Example 3: Inhibition of systemic and intestinal inflammatory factor secretion by Britannine
[0036] Serum was extracted from each group of mice, and the concentrations of IL-6, TNF-α, and IL-1β were detected using an ELISA kit. RNA was extracted from mouse colorectal tissue and two intestinal epithelial cell lines, and the mRNA levels of the above inflammatory factors were detected by qRT-PCR.
[0037] The results are as follows Figure 3 As shown, Britannin potently inhibits the overexpression of pro-inflammatory factors.
[0038] Example 4: Verification of the mechanism by which Britannin exerts its effects through the TRAF6 / NF-κB pathway
[0039] Western blotting was used to detect the phosphorylation level of p-P65, a key protein in the NF-κB pathway, in in vivo mouse animal model tissues and in vitro cell models.
[0040] The results are as follows Figure 4 As shown, Britannin significantly downregulates LPS-induced abnormal upregulation of p-P65, confirming that it improves septic intestinal injury by inhibiting TRAF6-mediated NF-κB pathway activation.
[0041] Example 5: Britannin exerts its anti-inflammatory effect by inhibiting macrophage M1 polarization.
[0042] RAW264.7 mouse macrophages were cultured in DMEM complete medium (10% fetal bovine serum, 100 U / mL penicillin, 100 μg / mL streptomycin) at 37°C and 5% CO2 until the cells reached 70%-80% confluence for the experiments.
[0043] The experiment was divided into 4 groups: ① Control group: treated with an equal amount of PBS solvent;
[0044] ②DMSO group: 1μg / mL LPS stimulation + equal volume of solvent DMSO;
[0045] ③ 1μM Group B: After pretreatment with 1μM Britannin for 1 h, 1μg / mL LPS was added for M1 polarization induction, and the mixture was incubated together for 24 h.
[0046] ④ 5μM Group B: After pretreatment with 1μM Britannin for 1 h, 1μg / mL LPS was added for M1 polarization induction, and the mixture was incubated together for 24 h.
[0047] Macrophages from each group were collected, and total RNA was extracted using the Trizol method. RNA concentration and purity were measured. An equal amount of RNA was reverse transcribed to synthesize cDNA. The mRNA expression level of the M1 marker gene iNos was detected by qRT-PCR, with Gapdh as an internal reference gene. -ΔΔCt The relative expression levels of each gene are calculated using this method.
[0048] The results are as follows Figure 5As shown, Britannin significantly inhibits the polarization of RAW264.7 macrophages to M1, confirming that it improves septic intestinal injury by inhibiting TRAF6-mediated NF-κB pathway activation.
[0049] Example 6: Britannin exerts its anti-inflammatory effect by promoting M2 polarization in macrophages.
[0050] RAW264.7 macrophages were cultured in DMEM complete medium (10% fetal bovine serum, 100 U / mL penicillin, 100 μg / mL streptomycin) at 37°C and 5% CO2. Experiments were conducted when the cell confluence reached 70%-80%.
[0051] The experiment was set up with 4 groups:
[0052] ① Control group (Ctrl): Add an equal volume of PBS solution;
[0053] ②DMSO group: Macrophages were stimulated to polarize to the M2 phenotype by adding 20 ng / mL IL-4 + 20 ng / mL IL-13 + equal volume of solvent DMSO;
[0054] ③ 1μM Group B: After pretreatment with 1μM Britannin for 1 h, 20 ng / mL IL-4 and 20 ng / mL IL-13 were added and incubated together for 24 h to promote M2 polarization;
[0055] ④ 5μM Group B: After pretreatment with 5μM Britannin for 1 h, 20 ng / mL IL-4 and 20 ng / mL IL-13 were added and incubated together for 24 h to promote M2 polarization.
[0056] Macrophages from each group were collected, and total RNA was extracted using the Trizol method. RNA concentration and purity were then measured. An equal amount of RNA was reverse transcribed to obtain cDNA. The mRNA expression level of the M2 biomarker gene Arg1 was detected by qRT-PCR, using Gapdh as an internal control gene. -ΔΔCt The relative expression levels of each gene are calculated using this method.
[0057] The results are as follows Figure 6 As shown, Britannin significantly promotes the polarization of RAW264.7 macrophages to M2, confirming that it improves septic intestinal injury by inhibiting TRAF6-mediated NF-κB pathway activation.
[0058] The above are merely preferred embodiments of the present invention and are not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.
Claims
1. The application of Inula japonica lactone in the preparation of drugs for treating septic intestinal injury, wherein the molecular formula of Inula japonica lactone is C 19 H 26 O7, chemical formula is .
2. The application according to claim 1, characterized in that, Sepsis-related intestinal injury refers to damage to the intestinal barrier function caused by sepsis.
3. The application according to claim 1, characterized in that, The drug is a preparation made with an effective dose of Inula japonica lactone as the active ingredient and pharmaceutically acceptable excipients.
4. The application according to claim 3, characterized in that, The drug is combined with antibiotics, anti-inflammatory drugs, or enteral nutrition protectants to form a compound preparation.
5. The application according to claim 3, characterized in that, The formulation is selected from at least one of the following formulations: oral, tablet, injection, or capsule.
6. The application according to claim 3, characterized in that, The preparation is an injectable formulation, and the administration method includes intravenous injection or subcutaneous injection.
7. A pharmaceutical composition for treating septic intestinal injury, characterized in that, It contains an effective dose of Inula japonica lactone as its active ingredient, wherein the molecular formula of Inula japonica lactone is C 19 H 26 O7, chemical formula is .
8. The application of Inula japonica lactone in the preparation of drugs that inhibit the expression of pro-inflammatory factors IL-6, TNF-α, and IL-1β, characterized in that, The molecular formula of Inula japonica lactone is C 19 H 26 O7, chemical formula is .
9. The application of Inula japonica lactone in the preparation of TRAF6-NF-κB signaling pathway inhibitors, characterized in that, The molecular formula of Inula japonica lactone is C 19 H 26 O7, chemical formula is This is achieved by inhibiting p-P65 expression and promoting ZO-1 expression.