Application of melatonin in anti-inflammatory intervention of bilirubin encephalopathy
By using melatonin for anti-inflammatory intervention in bilirubin encephalopathy, inhibiting the NLRP3 inflammasome and reducing the release of pro-inflammatory factors, and regulating the PI3K/AKT signaling pathway, the problem of lack of specific treatment for bilirubin encephalopathy inflammation in existing technologies has been solved, achieving a safe and effective neuroprotective effect.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- CHONGQING MEDICAL UNIVERSITY
- Filing Date
- 2026-02-03
- Publication Date
- 2026-05-26
AI Technical Summary
There is a lack of specific drug interventions for secondary brain damage caused by bilirubin encephalopathy. Current clinical interventions mainly focus on reducing bilirubin levels, while the therapeutic effect on neuroinflammation is limited.
Melatonin was used as the drug and administered via intraperitoneal injection at a dose of 10-20 mg/kg body weight, preferably 15 mg/kg body weight. It inhibited NLRP3 inflammasome activation, reduced GSDMD cleavage, decreased the levels of pro-inflammatory factors TNF-α, IL-6 and IL-1β, and regulated the PI3K/AKT signaling pathway to exert an anti-inflammatory effect.
It provides a safe and effective anti-inflammatory intervention strategy in addition to lowering bilirubin, which significantly reduces the inflammatory response of bilirubin encephalopathy, protects neurons, and improves the safety and effectiveness of clinical application.
Smart Images
Figure CN122075484A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of melatonin, and more particularly to the application of melatonin in anti-inflammatory intervention for bilirubin encephalopathy. Background Technology
[0002] Bilirubin encephalopathy (BE) is a serious central nervous system complication of neonatal hyperbilirubinemia. Neurological damage is caused not only by the direct toxicity of bilirubin, but also by neuroinflammation, which is considered a significant amplifying factor. Unconjugated bilirubin entering brain tissue can trigger glial cell damage and overactivation, inducing an inflammatory cascade that produces pro-inflammatory factors such as TNF-α, IL-1β, and IL-6. Studies have shown that bilirubin can upregulate the expression of the NLRP3 inflammasome and IL-1β in microglia, activating the "NLRP3 inflammasome-caspase-1-IL-1β" pathway and exacerbating inflammation and cell damage. Currently, clinical interventions for BE primarily focus on reducing bilirubin levels (e.g., phototherapy, exchange transfusion), but specific drug interventions for inflammation-induced secondary brain damage are lacking.
[0003] Melatonin, an endogenous indoleamine, is characterized by its high lipid solubility, easy crossing of the blood-brain barrier, and broad target range, demonstrating neuroprotective potential in perinatal / neonatal brain injury studies. Animal experiments have shown that melatonin can reduce bilirubin-induced cerebral edema and neuronal damage, regulate the PI3K / AKT signaling pathway, and possess anti-inflammatory effects, such as inhibiting NLRP3 inflammasome activation, reducing caspase-1 activation and IL-1β release, and downregulating pro-inflammatory factors such as TNF-α and IL-6. Therefore, exploring the application of melatonin in BE anti-inflammatory intervention is of great significance.
[0004] Therefore, this invention proposes the application of melatonin in the anti-inflammatory intervention of bilirubin encephalopathy. Summary of the Invention
[0005] The purpose of this invention is to address the shortcomings of existing technologies by proposing the application of melatonin in the anti-inflammatory intervention of bilirubin encephalopathy.
[0006] To achieve the above objectives, the present invention adopts the following technical solution: The use of melatonin in the preparation of a drug for the prevention or treatment of bilirubin encephalopathy.
[0007] Preferably, the dosage of melatonin is 10-20 mg / kg body weight.
[0008] Preferably, the dosage of melatonin is 15 mg / kg body weight.
[0009] Preferably, the drug is administered via intraperitoneal injection.
[0010] Preferably, melatonin exerts its anti-inflammatory effect by inhibiting the activation of the NLRP3 inflammasome.
[0011] Preferably, melatonin exerts its anti-inflammatory effect by reducing the cleavage of GSDMD.
[0012] Preferably, melatonin exerts its anti-inflammatory effect by reducing the levels of pro-inflammatory factors TNF-α, IL-6, and IL-1β.
[0013] Preferably, the effects of melatonin are related to the PI3K / AKT signaling pathway.
[0014] Preferably, the drug further comprises a pharmaceutically acceptable carrier.
[0015] A method for preventing or treating bilirubin encephalopathy includes administering an effective amount of melatonin to an individual in need, wherein the dose of melatonin is 10-20 mg / kg body weight, preferably 15 mg / kg body weight, by intraperitoneal injection.
[0016] The beneficial effects of this invention are as follows: 1. This invention provides a novel anti-inflammatory intervention strategy in addition to bilirubin-lowering, which is highly safe and easy to apply in clinical practice. Attached Figure Description
[0017] Figure 1 This is a schematic diagram showing the results of detecting the levels of inflammation-related proteins and cytokines after establishing the BE model according to the present invention; Figure 2 This is a schematic diagram of the Western Blot results after the BE model is established according to the present invention; Figure 3 This is a schematic diagram of the ELISA results of medium-dose MT treatment in relieving inflammation induced in the BE model according to the present invention; Figure 4 A schematic diagram of Western Blot results for the medium-dose MT treatment of the present invention to alleviate inflammation induced in the BE model; Figure 5 Venn diagram of 25 drug targets for chemotherapy-induced peripheral neuropathy, which are potential targets for network pharmacology analysis of the present invention; Figure 6 This invention presents a "drug-target" network diagram obtained by importing the intersection targets into Cytoscape 3.9.1 for visualization processing. Figure 7 The present invention utilizes the DAVID database to perform GO functional enrichment analysis on candidate targets and generate bar charts. Figure 8 KEGG pathway enrichment analysis and bubble diagram plotting were performed for this invention; Figure 9This is a schematic diagram of the Western blot results of the present invention. Detailed Implementation
[0018] The technical solution of the present invention will be further described in detail below with reference to specific embodiments.
[0019] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installation," "connection," "linking," and "setting" should be interpreted broadly. For example, they can refer to a fixed connection or setting, a detachable connection or setting, or an integral connection or setting. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.
[0020] Example 1: Establishment of BE model and dynamic analysis of inflammation time Animals: 7-day-old SD rats were provided by the Experimental Animal Center of Chongqing Medical University and housed in a controlled environment. All experiments were approved by the ethics committee.
[0021] Establishment of the BE model: 15 μL of cerebrospinal fluid was aspirated from the cerebellomedullary cistern to avoid increasing intracranial pressure. Bilirubin solution (10 μg / g body weight) was injected into the cerebellomedullary cistern according to body weight, and the Sham group received an equal volume of sterile solvent.
[0022] Sample collection: Blood was collected from the heart at 12h, 24h and 48h after modeling under pentobarbital anesthesia, and the brain was collected after perfusion with normal saline.
[0023] ELISA assay: Serum TNF-α, IL-6, and IL-1β levels were detected using 96-well plates coated with antibodies. Results showed that inflammatory factors in the BE group peaked at 24 hours (P<0.001).
[0024] Western Blot: Brain tissue was lysed with RIPA lysis buffer, centrifuged, and the supernatant was collected. Protein concentration was measured by BCA, followed by SDS-PAGE electrophoresis. After transfer to a membrane, NLRP3 and N-GSDMD proteins were detected. The results showed that NLRP3 and N-GSDMD expression was strongest in the BE group at 24 h.
[0025] Example 2: Melatonin Dosage Optimization and Anti-inflammatory Effect Evaluation Animal grouping: Animals were randomly divided into the Sham group, BE group, BE + low-dose melatonin group (LM, 10 mg / kg), BE + medium-dose melatonin group (MM, 15 mg / kg), and BE + high-dose melatonin group (HM, 20 mg / kg). Melatonin solution preparation: Weigh 10 mg, 15 mg, and 20 mg of melatonin, dissolve each in 1 mL of anhydrous ethanol, and add 0.9% NaCl to 20 mL to obtain concentrations of 0.50 mg / mL, 0.75 mg / mL, and 1.00 mg / mL, respectively. Prepare freshly, protected from light, and administer intraperitoneally 30 minutes before modeling.
[0026] ELISA results showed that compared with the BE group, the levels of TNF-α, IL-6, and IL-1β in the medium-dose melatonin group (MM) were significantly decreased (P<0.05), approaching those in the Sham group. The improvement was less pronounced in the low-dose and high-dose groups.
[0027] Western Blot: The expression of NLRP3 and N-GSDMD was significantly reduced in the medium-dose melatonin group, suggesting that the medium dose is the optimal anti-inflammatory dose.
[0028] Example 3: Network pharmacology analysis of melatonin's target sites Target collection: Melatonin SMILES were obtained through PubChem and imported into databases such as SwissTargetPrediction, resulting in 100 MT targets.
[0029] Disease target screening: Using keywords such as "bilirubin encephalopathy" to search the GeneCards database, 1670 BE targets were obtained and duplicates were removed.
[0030] Intersection Targets: The intersection of MT and BE targets is taken to obtain 25 candidate targets. A Venn diagram is then drawn. Figure 5 ).
[0031] Protein-protein interaction network: Intersecting target points were imported into the STRING database (score > 0.4), visualized using Cytoscape, and topological parameters were calculated to obtain core target points.
[0032] GO and KEGG enrichment analysis: DAVID database analysis showed that GO enriched the target in signal transduction and PI3K / AKT pathways, etc.; KEGG enriched the target in the PI3K-AKT signaling pathway, etc. Figure 8 ).
[0033] Example 4: PI3K / AKT Pathway Validation Experiment Using the PI3K inhibitor LY294002: Prepare a working solution of LY294002 (10 mg / mL stock solution, plus PEG300, Tween80 and physiological saline), and administer intraperitoneally at a dose of 1.2 mg / kg, 30 min before melatonin treatment.
[0034] Animal groups: Sham group, BE group, BE + medium-dose melatonin group (MM), BE + MM + LY294002 group.
[0035] Western Blot: Compared with the BE group, the expression of NLRP3 and N-GSDMD was reduced in the MM group; after LY294002 intervention, the inhibitory effect of the MM group was partially offset, and NLRP3 and N-GSDMD remained at a high level, suggesting that the effect of melatonin depends on the PI3K / AKT pathway.
[0036] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.
Claims
1. The use of melatonin in the preparation of a medicament for the prevention or treatment of bilirubin encephalopathy.
2. The use of melatonin according to claim 1 in the preparation of a medicament for the prevention or treatment of bilirubin encephalopathy, characterized in that, The dosage of melatonin mentioned above is 10-20 mg / kg body weight.
3. The use of melatonin according to claim 2 in the preparation of a medicament for the prevention or treatment of bilirubin encephalopathy, characterized in that, The dosage of melatonin mentioned above is 15 mg / kg body weight.
4. The use of melatonin according to claim 1 in the preparation of a medicament for the prevention or treatment of bilirubin encephalopathy, characterized in that, The drug is administered via intraperitoneal injection.
5. The use of melatonin according to claim 1 in the preparation of a medicament for the prevention or treatment of bilirubin encephalopathy, characterized in that, The melatonin mentioned therein exerts its anti-inflammatory effect by inhibiting the activation of the NLRP3 inflammasome.
6. The use of melatonin according to claim 1 in the preparation of a medicament for the prevention or treatment of bilirubin encephalopathy, characterized in that, The melatonin mentioned therein exerts its anti-inflammatory effect by reducing the cleavage of GSDMD.
7. The use of melatonin according to claim 1 in the preparation of a medicament for the prevention or treatment of bilirubin encephalopathy, characterized in that, The melatonin mentioned above exerts its anti-inflammatory effect by reducing the levels of pro-inflammatory factors TNF-α, IL-6 and IL-1β.
8. The use of melatonin according to claim 1 in the preparation of a medicament for the prevention or treatment of bilirubin encephalopathy, characterized in that, The effect of melatonin is related to the PI3K / AKT signaling pathway.
9. The use of melatonin according to claim 1 in the preparation of a medicament for the prevention or treatment of bilirubin encephalopathy, characterized in that, The drug also contains a pharmaceutically acceptable carrier.
10. A method for preventing or treating bilirubin encephalopathy, characterized in that, This includes administering an effective amount of melatonin to an individual in need, wherein the dose of melatonin is 15 mg / kg body weight, administered via intraperitoneal injection.