New application of saikosaponin C in inhibiting neuroinflammation

A technology of neuroinflammation and saikosaponin, applied in the field of medicine, can solve problems such as saikosaponin C that has not been seen yet

Inactive Publication Date: 2019-07-05
NINGXIA MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, reports on the pharmacological activity of saikosapogenin C are very rare, and there

Method used

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  • New application of saikosaponin C in inhibiting neuroinflammation
  • New application of saikosaponin C in inhibiting neuroinflammation
  • New application of saikosaponin C in inhibiting neuroinflammation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: Griess colorimetric method investigates the inhibitory effect of saikogenin C on LPS-induced microglia nitric oxide (NO) release;

[0030] Cell line: mouse microglial cell line BV-2;

[0031] Drugs: LPS; saikosaponin C; minocycline (MINO).

[0032] method:

[0033] (1) Culture of mouse microglial cell line BV-2:

[0034] The cell culture solution was prepared based on DMEM medium, containing 10% FBS, 100 U penicillin and 100 U streptomycin, and 50 μM 2-mercaptoethanol (all final concentrations). at 5% CO 2 , under the condition of 37 ℃, the BV-2 microglial cells were divided into about 4×10 5 The cell density of cells / ml was cultured in a cell culture incubator, and the growth of the cells was observed regularly. When the area of ​​the cell adherence accounted for about 50-60% of the bottom area of ​​the culture flask, the cells were digested with trypsin and passaged, and then continued to culture. Take 3-7 generations of cells for experiments.

[0...

Embodiment 2

[0042] Example 2: The DCFH oxidation method was used to investigate the effect of saikosapogenin C on the release of reactive oxygen species (ROS) from microglial cells induced by LPS;

[0043] Cell line: mouse microglial cell line BV-2;

[0044] Drugs: LPS; saikosaponin C; MINO.

[0045] Experimental principle: DCFH-DA (2′,7′-dichlorodihydrofluororescein diacetate) can enter the cell and undergo a deacetylation reaction to generate DCFH. DCFH is a non-fluorescent substance, which can be oxidized by intracellular ROS to generate fluorescent substance DCF, and the amount of ROS release can be reflected by detecting the fluorescence intensity. The specific operation steps are as follows: DCFH-DA was dissolved in pure methanol to prepare a mother solution with a concentration of 10 mM, and the mother solution was diluted 500 times with Hank's balanced salt solution (HBSS) to a final concentration of 20 μM before use. Take the BV-2 cells after drug treatment, suck the supernatant,...

Embodiment 3

[0049] Example 3: The effect of saikosapogenin C on the release of LPS-induced microglial tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) by ELISA;

[0050] Cell line: mouse microglial cell line BV-2;

[0051] Drugs: LPS; saikosaponin C; MINO.

[0052] Detection method: Take BV-2 cells in the logarithmic growth phase, inoculate the cells in a 6-well plate with fresh serum-free DMEM medium, and the cell density is 3×10 5 cells / ml, inoculum volume 600μl / well, placed in an incubator at 37°C, 5% CO 2 cultivated under conditions. After the cells adhered to the wall for 3 hours, the culture solution was carefully sucked out, and according to the experimental group, the cells were replaced with different drugs prepared in serum-free DMEM culture solution to incubate the cells. At the same time, a blank control group was set up, and each group was set with 3 replicate wells, and the drug addition to the cells continued. After culturing for 2-4 hours, collect the supernat...

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Abstract

The invention discloses a new application of saikosaponin C in inhibiting neuroinflammation. Pharmacological experiments prove that saikosaponin C can significantly inhibit LPS-induced release of microglial inflammatory mediators NO, ROS, TNF-alpha and IL-1beta, and can significantly inhibit neuroinflammatory activity. Therefore, the compound can be used for preparing drugs for preventing and/or treating neuroinflammation and related central neurodegenerative diseases, and has higher clinical application value and development prospect.

Description

[0001] Technical field: [0002] The invention relates to the technical field of medicine, in particular to the use of saikosaponin C in inhibiting neuroinflammation. [0003] Background technique: [0004] Neurodegenerative diseases (neurodegeneration disorders, ND) are a group of chronic progressive neurological diseases based on primary neuron degeneration, mainly including Alzheimer's disease (Alzheimer's disease, AD), Parkinson's disease (Parkinson's disease) , PD), multiple sclerosis (multiple sclerosis, MS), Huntington's disease (Huntington's disease, HD) and so on. With the aging of the world's population, the incidence of neurodegenerative diseases is on the rise. For example, the prevalence of AD in people over 60 years old in my country is 5.1%, and that in people over 85 years old is 30%. There are more than 8 million people, and there are about 35 million AD patients in the world. The prevalence of PD is second only to AD, and mainly occurs in people over middle a...

Claims

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Application Information

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IPC IPC(8): A61K31/56A61P25/00A61P25/14A61P25/16A61P25/28A61P29/00
CPCA61K31/56A61P25/00A61P25/14A61P25/16A61P25/28A61P29/00
Inventor 李娟姚遥陈菲李玮琦
Owner NINGXIA MEDICAL UNIV
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