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Application of scpoletin to preparation of medicament for preventing and treating autoimmune diseases

An autoimmune disease, scopolamine technology, applied in the field of biomedicine, can solve the problem that the influence of innate immune cell regulatory characteristics of pathogenic progression is poorly understood and other issues

Inactive Publication Date: 2019-03-08
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, little is known about the regulatory properties of scopolamine on innate immune cells such as DCs and their impact on the pathogenic progression of MS and EAE

Method used

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  • Application of scpoletin to preparation of medicament for preventing and treating autoimmune diseases
  • Application of scpoletin to preparation of medicament for preventing and treating autoimmune diseases
  • Application of scpoletin to preparation of medicament for preventing and treating autoimmune diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Preventive administration of scopoletin relieves central and peripheral inflammation in EAE mice

[0023] 1. Preparation of EAE mouse model and preventive administration by intraperitoneal injection

[0024] Eight-week-old female C57BL / 6 mice were purchased from Fourth Military Medical University (Xi'an, China). All experimental procedures and protocols were approved by the Institutional Animal Care and Use Committee of Shaanxi Normal University and performed in accordance with approved institutional guidelines and regulations. Mice were bred under standard conditions (23±2°C) in the Animal Breeding Room of Shaanxi Normal University. To induce EAE, 200 μL of myelin oligodendrocyte glycoprotein peptide (MOG 35-55 ), 50% volume complete Freund's adjuvant, and 5 mg / mL Mycobacterium tuberculosis H37Ra (Sigma-Aldrich) were immunized subcutaneously at two sites on the back. And all mice were intraperitoneally injected with 200 ng of pertussis toxin (supplied by Sigma-Aldri...

Embodiment 2

[0041] Scopoletin inhibits the activation of mouse BM-DCs in vitro

[0042] 1. Experimental Treatment

[0043] Take two normal C58BL / 6 female mice about 8 weeks old, take out the femur and tibia under aseptic conditions, remove the surrounding tissue, draw PBS into the bone marrow cavity with a sterile syringe, wash out the bone marrow repeatedly and place it in a petri dish on ice. Collect bone marrow suspension, centrifuge and count after nylon mesh filtration, according to 1.5×10 6 cells / mL were seeded into a Petri dish and placed in CO at 37°C 2 incubator.

[0044] The cells were divided into two groups, one group was only stimulated with 100ng / mL LPS in vitro, and the other group was stimulated with 100ng / mL LPS in vitro while adding 100μM scopolamine, and the cells were collected after 72 hours for staining and flow cytometry detection. And the supernatant of DCs was used to measure the concentration of IFN-γ, IL-17, GM-CSF, IL-1β, IL-6 and IL-23 using ELISA kit (R&D ...

Embodiment 3

[0051] Scopolamine inhibits NF-κB signaling during LPS-stimulated DCs maturation in vitro

[0052] 1. Experimental Treatment

[0053] The mature BM-DCs obtained in Example 2 above were planted in a 6-well plate at a density of 1.5×10 6 cells / mL, stimulated with 100ng / mL LPS to induce the activation of BM-DCs, and added 100μM scopoletin at the same time. After 18 hours, proteins were extracted from BM-DCs. Protein concentrations were determined using the Pierce™ BCA Protein Assay Kit (Thermo Fisher Scientific, Rockford, MA). Equal amounts of protein from BM-DCs were added to sodium dodecyl sulfate polyacrylamide gel electrophoresis (sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE), and transferred to polyvinylidene fluoride (polyvinylidene fluoride, PVDF) On the membrane, after blocking with 5% skimmed milk in Tris-buffered saline (TBS), incubate the membrane with primary antibodies at 4°C overnight, and the primary antibodies are JNK, p-JNK, NF-κB, p-NF ...

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Abstract

The invention discloses application of scpoletin to preparation of a medicament for preventing and treating autoimmune diseases. The invention shows that the scpoletin remarkably retards human multiple sclerosis (MS) as well as the central nervous system inflammation and demyelinating disease of an animal model experimental autoimmune encephalomyelitis (EAE) mouse; and the expression amounts of MHC II, CD80 and CD86 on the surfaces of dendritic cells (DCs) are reduced by two ways in vivo and in vitro to influence the infiltration and differentiation of encephalitogenic Th1 / Th17 cells, and theimmunosuppressive action is brought into play specifically through an NF-kappaB signal transduction pathway to relieve the clinical symptoms of EAE, thereby proving that the scpoletin has the pharmacological actions of lowering the antigen-presenting ability of the DCs and inhibiting EAE pathogenetic conditions. According to the invention, an important value is provided for the development of natural compounds and new anti-inflammatory agents, and the treatment of the autoimmune diseases.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to the application of scopolamine in the preparation of medicines for preventing and / or treating autoimmune diseases, especially the application of scopolamine in the preparation of medicines for preventing and / or treating autoimmune diseases. Background technique [0002] Autoimmune diseases refer to diseases caused by the body's immune response to self-antigens, resulting in damage to its own tissues. Antibodies of the immune system in the human body are originally designed to attack and eliminate foreign antigens or abnormal cells in the body, which is a physiological mechanism to protect the body. But in some cases, the immune system may produce antibodies against normal cells in the body, causing excessive inflammation or tissue damage, which in turn affects health and causes diseases, including type 1 diabetes, Graves' disease, multiple sclerosis syndrome, psor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/37A61P37/02
CPCA61K31/37A61P37/02
Inventor 张媛申沛鑫李星张菲赵莉韩娟娟
Owner SHAANXI NORMAL UNIV
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