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Application of monomer luteolin coming from vegetables and fruits and Chinese herbal medicine

A technology of luteolin and Chinese herbal medicine, applied in the field of biomedicine, can solve problems affecting the polarization of macrophage inflammation

Inactive Publication Date: 2019-05-21
JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Literature (Wang SX,Cao M,Xu SH,Zhang JM,Wang ZG,Mao XD,Liu C,Yao XM.Effect of luteolin on inflammatory responses inRAW264.7macrophages activated with LPS and IFN-γ[J].J Funct Foods,2017 ,32:123-130.) reported that luteolin can significantly reduce the inflammatory response of activated macrophages and down-regulate the expression of inflammatory factors such as IL-6, IL-1β and TNF-α, but whether luteolin affects The inflammatory polarization of macrophages has not been reported so far

Method used

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  • Application of monomer luteolin coming from vegetables and fruits and Chinese herbal medicine
  • Application of monomer luteolin coming from vegetables and fruits and Chinese herbal medicine
  • Application of monomer luteolin coming from vegetables and fruits and Chinese herbal medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Embodiment 1: MTT method detects cell viability

[0016] Cells growing in logarithmic phase were taken to prepare single cell suspension, and cultured overnight in 96-well plates. LPS and IFN-γ stimulated cells to induce M1-type polarization, and IL-4 stimulated to induce M2-type polarization. At the same time, M1 cells were treated with luteolin at final concentrations of 5, 10, 20, 40, and 80 μmol / L. After 24 hours, Add 20 μL of MTT (5 g / L, PBS) to each well, discard the supernatant after 4 hours, add 150 μL of DMSO to each well, measure the absorbance (A value) after the crystals are completely dissolved, and calculate the growth rate. According to the above growth curve, select the corresponding luteolin concentration for subsequent steps.

[0017] Growth rate=(A value of experimental group / A value of control group)×100%.

[0018] Luteolin at concentrations lower than 20 μmol / L had no effect on the viability of polarized cells. MTT results showed that IL-4-induce...

Embodiment 2

[0019] Embodiment 2: ELISA detection supernatant cytokine content

[0020] 2×10 cells per well 5 cells / mL density, after overnight culture, LPS and IFN-γ or IL-4 induced polarization, and then added different doses of luteolin. After 24 hours, the supernatant was collected, and the levels of IL-6 and TNF-α were detected. The specific operation was performed according to the ELISA kit detection instructions.

[0021] IL-6 and TNF-α secreted by activated M1 cells were significantly up-regulated, compared with the normal control group, there was a significant difference; after luteolin treatment, the secretion of cytokines decreased, compared with the M1 group, there was a significant difference; IL- 4 The IL-6 and TNF-α secreted by the activated M2 cells were significantly down-regulated, compared with the M1 group, there was a significant difference, and there was no significant difference compared with the control group. The result is as follows:

[0022] group

...

Embodiment 3

[0024] Example 3: Fluorescent quantitative PCR detection of expression of inflammatory factors

[0025] Treat the cells overnight in the same way as in Example 2, collect the cells, extract the total RNA by Trizol method and reverse transcribe it into cDNA, amplify the target gene by fluorescent quantitative PCR, and calculate the relative expression levels of inflammatory factors and anti-inflammatory factors.

[0026] After the intervention of different concentrations of luteolin in M1 polarized macrophages, the secretion of M1-type inflammatory factors such as iNOS, IL-6 and IL-1β was significantly down-regulated, which was significantly different from the M1 group; M2-type related The expression of anti-inflammatory molecules such as Arg1, CD206 and CD163 was up-regulated in a concentration-dependent manner, and there was a significant difference compared with the M1 group. See figure 2 .

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Abstract

The invention discloses application of monomer luteolin coming from vegetables and fruits and Chinese herbal medicine, and particularly relates to the influence and mechanism of the luteolin coming from vegetables and fruits and Chinese herbal medicine on the macrophage polarization mode and new application of the luteolin in the anti-inflammation aspect. Different polarization modes of macrophages have important significance in development and prognosis of tumors, angiocardiopathy, nephrosis, inflammations and the like. Induction of M1-type polarization of the macrophages is stimulated with bacterial endotoxin lipopolysaccharide and interferon-gamma; induction of M2-type polarization is stimulated with IL-4; after the luteolin is adopted for treatment, the morphological change of the macrophages and expression of marking molecules of the macrophages after M1-type polarization and M2-type polarization are observed; an immunoblotting technology is adopted for detecting the expression ofa protein channel. It is found that the luteolin can down-regulate M1-type pro-inflammatory cytokines and up-regulate M2-type pro-inflammatory cytokines; meanwhile, protein p-STAT3 is horizontally down-regulated, and p-STAT6 is up-regulated, that is to say, the luteolin regulates the polarization of the M1 / M2 macrophages through the protein channel STATs, and thus the anti-inflammatory effect isachieved.

Description

technical field [0001] The invention belongs to the application of luteolin derived from natural edible plants, specifically the application of luteolin in regulating the polarization of mouse macrophages and anti-inflammation, and belongs to the field of biomedicine. Background technique [0002] Severe pathogenic infection can lead to sepsis, which is dangerous and develops rapidly. In severe cases, it can develop into multiple organ failure or even life-threatening. This process is often accompanied by severe inflammatory reactions. Inflammation is an immune defense response of the body against pathogenic infection, which is beneficial to clear pathogenic bacteria and control infection, but excessive inflammatory response can cause pathological damage. Inflammatory factors secreted as activation of inflammatory cells against infection can also activate quiescent tumor cells to grow and evolve into malignancies. Therefore, it is crucial to study the precise regulation of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/352A61P29/00A61P35/00A61P31/00A61P13/12A61P9/10A61P3/10
Inventor 王书侠姚孝明张振海徐书杭曹萌施健丰
Owner JIANGSU PROVINCE INST OF TRADITIONAL CHINESE MEDICINE
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