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Application of scutellarin to treatment of microglia-mediated diseases

A technology of microglia and scutellarin, applied in the field of scutellarin

Inactive Publication Date: 2011-04-06
TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, scutellarin has not been reported in inhibiting microglia-mediated neurotoxicity, and preventing and treating Parkinson's disease, multiple sclerosis, Huntington's disease, encephalitis, Gujaré's disease, etc.

Method used

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  • Application of scutellarin to treatment of microglia-mediated diseases
  • Application of scutellarin to treatment of microglia-mediated diseases
  • Application of scutellarin to treatment of microglia-mediated diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] Example 1: Effect of scutellarin on secretion of inflammatory mediators by microglia

[0071] Complete DMEM medium (containing 10% fetal bovine serum, 100 U / mL penicillin and 100 μg / mL streptomycin) was used to culture mouse microglial cell line BV-2 and primary cultured rat microglial cells. Adjust cell concentration to 1 x 10 5 / mL, inoculate into 48-well culture plate overnight, add different concentrations of scutellarin (final concentration is 2, 10, 50 μmol / L) and pre-incubate for 0.5h, then add endotoxin (LPS, final concentration 0.1 μg / mL) Stimulate. Experimental groups: normal control group, LPS group and scutellarin plus LPS group. The cells were cultured for 24 hours (or 8, 12, 16, 20, 24 hours) after adding LPS, and the cell culture supernatant of each treatment group was collected, and the concentration of NO was detected by Greiss method, and the contents of cytokines TNFα and IL-1β were detected by Elisa method. The operation steps were completed acc...

Embodiment 2

[0074] Example 2: Scutellarin on the expression of iNOS, TNFα, IL-1β mRNA in microglial cells Impact

[0075] The mouse microglial cell line BV-2 and primary rat microglial cells were planted in 12-well culture plates overnight, pre-incubated with scutellarin (50 μmol / L) for 0.5 h, and then added LPS (0.1 μg / L mL) to stimulate for 8h. Wash once with PBS, and extract total cellular RNA with Trizol reagent. Samples were taken for ultraviolet detection with wavelengths of 260nm and 280nm, and the purity of total RNA was analyzed and quantified. Total RNA (0.2 μg) was reverse-transcribed into cDNA (10 μl system) using TaqMan Reverse Transcription Reagents Kit (Applied Biosystems) and oligod(T)16 primer. 0.5 μl of the cDNA product was used as template for quantitative PCR amplification. The PCR amplification reaction used SYBR Green PCR Master Mix reagent kits (Applied Biosystems), and corresponding specific primers:

[0076] mouse-GAPDH

[0077] Forward: CTTCACCACCATGGAGA...

Embodiment 3

[0102] Example 3: Effect of scutellarin on microglia ROS

[0103] Using DCFH-DA as a fluorescent probe indicating the level of intracellular ROS, the effect of scutellarin on the release of ROS from mouse BV-2 cells was studied. DCFH-DA itself does not emit fluorescence. After entering the cell, it is oxidized by ROS in the cell and emits fluorescence, thereby indicating the level of ROS in the cell. BV-2 cells were seeded in 48-well plates or 24-well plates (including coverslips). After the BV-2 cells were treated with LPS and / or scutellarin for 4 hours, the supernatant was discarded, and the cells were washed with PBS. Add DMEM medium containing DCFH-DA (10 μmol·L-1) probe to each well and continue to incubate for 30 min. Discard the supernatant, wash away the probes that have not entered the cells with PBS, add 200 μL PBS to each well, read the fluorescence value on a microplate reader, or take pictures with a laser confocal scanning microscope, the excitation wavelengt...

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Abstract

The invention relates to application of scutellarin shown as a formula I to the treatment of microglia-mediated diseases. Typically, the invention relates to the application of the scutellarin to the preparation of medicaments for treating and / or preventing the microglia-mediated diseases, in particular to the application of the scutellarin to the preparation of medicaments for treating and / or preventing microglia-mediated neurotoxicity-caused neural immune inflammatory diseases. The scutellarin has the novel functions of remarkably inhibiting the generation of inflammatory mediators, namely nitric oxide (NO), a tumor necrosis factor (TNF) alpha and interleukin (IL)-1beta which activate microglia, inhibiting the expression of inducible nitric oxide synthase (iNOS), TNF alpha and IL-1beta messenger ribonucleic acid (mRNA), inhibiting the generation of reactive oxygen species in the microglia, reducing the activation of nuclear factor (NF)-kappa B and remarkably alleviating the cytotoxicity of the microglia to neurons. Therefore, the scutellarin can be applied to the prevention and treatment of brain immune inflammatory related diseases such as Parkinson's disease and the like. The formula I is shown in the specifications.

Description

technical field [0001] The present invention relates to a new application of scutellarin, in particular to the treatment and / or prevention of neuroimmune inflammatory diseases such as Parkinson's disease, multiple sclerosis and Huntington's disease caused by scutellarin mediated by neurotoxic effects of microglial cells New uses for diseases such as , encephalitis, and Gujaré's disease. Background technique [0002] Parkinson's disease (Parkinson's disease) is a common movement disorder in middle-aged and elderly people, also known as parkinsonism. body formation. The main clinical manifestations are resting tremor, muscle rigidity, bradykinesia and other symptoms. The etiology of Parkinson's disease and the precise mechanisms that cause degeneration of dopaminergic neurons have not yet been fully elucidated. Parkinson's disease is still an incurable progressive disease, and the current treatment is essentially to improve the patient's quality of life and work ability. W...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7048A61P25/16A61P25/00A61P25/14
Inventor 胡利民王少峡高秀梅康立源王虹
Owner TIANJIN UNIV OF TRADITIONAL CHINESE MEDICINE
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