Application of carnosol in preparation of medicines for preventing and treating experimental autoimmune encephalomyelitis

An autoimmune, carnosol-based technology for biomedical and pharmaceutical applications

Active Publication Date: 2018-10-23
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, it is a natural active compound favored by new drug researchers in recent years, but whether it can p

Method used

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  • Application of carnosol in preparation of medicines for preventing and treating experimental autoimmune encephalomyelitis
  • Application of carnosol in preparation of medicines for preventing and treating experimental autoimmune encephalomyelitis
  • Application of carnosol in preparation of medicines for preventing and treating experimental autoimmune encephalomyelitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] 1. Experimental treatment

[0046] (1) EAE model preparation

[0047] Eight-week-old female C57BL / 6J mice were purchased from the Air Force Military Medical University of the Chinese People's Liberation Army (Xi'an, China). myelin oligodendrocyte glycoprotein peptide 35-55 (MOG 35-55 ), purchased from Genescript Company, pertussis toxin, purchased from Sigma-Aldrich Company, complete Freund's adjuvant containing Mycobacterium tuberculosis, purchased from BD Difco Company.

[0048] Dissolve MOG with PBS 35-55 The polypeptide is then mixed with an equal volume of complete Freund's adjuvant (containing 5 mg / ml Mycobacterium tuberculosis), and pushed with a glass syringe until a water-in-oil white antigen emulsion is formed.

[0049] Mice were immunized at two sites on the back, and diluted pertussis solution (200 ng / mouse) was injected intraperitoneally on the day of immunization and 2 days later, respectively.

[0050] Grouping and dosing:

[0051] ①PBS-treated cont...

Embodiment 2

[0071] 1. Experimental Treatment

[0072] The same modeling administration as in Example 1, MNC was prepared from the spleen and lymph nodes of IL-17A-IRES-GFP mice (C57BL / 6J) on the 10th day after modeling to study the pathogenicity of carnosol on Th17 cells Sexuality, cells were cultured with PBS and carnosol under Th17 differentiation conditions, and treated with MOG 35-55 (25 μg / ml), IL-2 (2ng / ml) and IL-23 (10ng / ml) stimulation. After three days in culture, CD4+ T cells were isolated. The cells (4×10 6 cells) injected into C57BL / 6J recipient mice. After 20 days, the mice were sacrificed, and different groups of brain tissues were collected for immunohistochemical staining.

[0073] 2. Experimental results

[0074] (1) Compared with the PBS treatment group, the Th17 cells treated with carnosol significantly reduced the severity of clinical disease after transfer (p Figure 10 ).

[0075] (2) Immunofluorescence staining results of the mice sacrificed after the 20th ...

Embodiment 3

[0077] 1. Experimental Treatment

[0078] (1) C57BL / 6J mice aged 8-10 weeks were treated with MOG 35-55 After immunization, the mice were treated from day 25 and injected with PBS and carnosol for clinical scoring.

[0079] (2) On the 60th day of treatment, the spinal cord and lumbosacral enlargement of PBS and EAE mice were excised for immunohistochemistry and MBP staining, and quantitatively analyzed with Imag-Pro.

[0080] (3) Staining of pro-inflammatory (M1) and immunomodulatory (M2) markers of infiltrating macrophages and innate microglia in the mouse spinal cord to assess the effect of carnosol on these cells in the spinal cord.

[0081] (4) Prepare primary microglial cells from newborn C57BL / 6 mice, stimulate with lipopolysaccharide (LPS, 100ng / ml) and treat with different concentrations of carnosol for 2 days, and detect TNF in the supernatant by ELISA -α expression, and the cells were collected to detect the expression of related genes by real-time fluorescent quan...

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Abstract

The invention discloses application of carnosol in preparation of medicines for preventing and treating experimental autoimmune encephalomyelitis. Whole animal model experiments verifies that the carnosol not only can effectively delay the occurrence and development of the experimental autoimmune encephalomyelitis, significantly reduces the infiltration of peripheral inflammatory cells into the central nervous system and relieves the degree of demyelination, but also can promote the transformation of infiltrating macrophages and intrinsic microglial proinflammatory phenotypes to an M2 phenotype with an immunoregulatory function. Furthermore, the carnosol can inhibite Th17 cell differentiation and STAT3 phosphorylation and block NF-kappa B nuclear translocation of a transcription factor. Therefore, the carnosol has a great potential in the preparation of the medicines for preventing and treating the autoimmune diseases such as multiple sclerosis.

Description

technical field [0001] The invention belongs to the field of biomedicine and pharmaceutical applications, and in particular relates to the application of carnosol in the preparation of drugs for preventing and treating experimental autoimmune encephalomyelitis. Background technique [0002] Multiple sclerosis (Multiple Sclerosis, MS) and its animal model, Experimental Autoimmune Encephalomyelitis (EAE), is a kind of immune cell activated by itself that acts on the central nervous system (Central Nervous System, CNS). ) is a neurodegenerative disease characterized by inflammatory cell infiltration and demyelination. MS clinical symptoms mainly manifest as blurred vision, body numbness, and quadriplegia, etc. It occurs mostly in young people, and it is difficult to cure and has a high disability rate. It is currently distributed all over the world, and more than 2.5 million people have suffered from this disease , the incidence rate in our country also increases year by year,...

Claims

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Application Information

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IPC IPC(8): A61K31/366A61P37/02A61P25/00
Inventor 张媛鹿欣雨李星韩娟娟张菲赵莉
Owner SHAANXI NORMAL UNIV
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