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Application of hypholomethine B in preparation of immune activator

An immune activation and activity technology, applied in the direction of organic active ingredients, medical preparations containing active ingredients, antibacterial drugs, etc., can solve the problem of no relevant reports on immune regulation

Active Publication Date: 2019-04-16
ZHEJIANG CHINESE MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that Hypholomine B has strong free radical scavenging activity and sialidase inhibitory activity, so it has potential antioxidant and antiviral effects, while other research reports, especially immunomodulatory effects, have not been reported.

Method used

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  • Application of hypholomethine B in preparation of immune activator
  • Application of hypholomethine B in preparation of immune activator
  • Application of hypholomethine B in preparation of immune activator

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Effect of Hypholomine B on the proliferation of RAW264.7 cells

[0019] RAW264.7 cells (purchased from Shanghai Chinese Academy of Sciences Cell Bank) were incubated at 37°C with 5% CO in DMEM medium (Gibco) containing 10% fetal bovine serum and double antibodies (streptomycin 100 μg / mL and penicillin 100 U / mL). 2 cultured in an incubator. Take the RAW264.7 cells in the exponential growth phase, wash twice with 4°C pre-cooled PBS, collect the cells by pipetting, and culture them in DMEM containing 10% fetal bovine serum and double antibodies (streptomycin 100 μg / mL and penicillin 100 U / mL) Base diluted to 1 x 10 5 cells / mL, inoculate 96-well plate, 200 μL per well, 37°C, 5% CO 2 The incubator was incubated overnight. Add Hypholomine B at different final concentrations (0, 10, 20, 40, 60, 80, 100, 120 μg / mL) and continue to incubate for 44 hours, then add 10 μL of CCK8 reagent to each well, continue to incubate for 4 hours, measure the absorbance value at 4...

Embodiment 2

[0022] Example 2: Hypholomine B promotes secretion of INF-γ and other cytokines by mouse peritoneal macrophages and RAW264.7 cells

[0023] ICR mice were sacrificed by cervical dislocation, and aseptically collected peritoneal macrophages after soaking in 75% ethanol, and diluted with DMEM medium containing 10% fetal bovine serum and double antibodies (streptomycin 100 μg / mL and penicillin 100 U / mL) into 1×10 5 cells / mL, inoculate a 24-well plate, 0.5 mL per well; take RAW264.7 cells in the exponential growth phase, and use DMEM containing 10% fetal bovine serum and double antibodies (streptomycin 100 μg / mL and penicillin 100 U / mL) Medium diluted to 1 x 10 5 cells / mL, seed a 12-well plate, 1 mL per well, store at 37°C, 5% CO 2 Cultured overnight in the incubator; Hypholomine B with different final concentrations (10, 20, 40, 60 μg / mL) was added to continue the culture, and 1 μg / mL of LPS was used as a positive control, and an equal amount of medium was added as a negative co...

Embodiment 3

[0031] Embodiment 3: Hypholomine B promotes the phagocytic activity of macrophages to Escherichia coli

[0032] Take the supernatant of Example 2, and measure the phagocytosis activity of the cells by flow cytometry according to the fluorescence intensity after the cells phagocytize the bacteria, the results are as follows: figure 2 shown. The average fluorescence intensity of the normal group was (4075.17±253.36), and its peak value only shifted to the right by 3.8% after 1 μg / mL LPS stimulation for 12 hours, and the average fluorescence intensity was (4990.95±207.35). Showed very significant phagocytic activity; while Hypholomine B showed a significant dose effect, when treated at a concentration of 60 μg / mL, the peak shifted to the right by 17.2% as a whole, and the average fluorescence intensity was (11289.71±477.71), showing a significant promotion of phagocytosis effect.

[0033] Fluorescence photography results of confocal high-content imaging analysis system are as ...

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Abstract

The invention discloses an application of hypholomine B in the preparation of an immune activator, the hypholomine Bcan effectively promote the secretion of INF-gamma and other cytokines by macrophages, INF-gamma and other cytokines have strong antiviral, immunoregulatory and antitumor activities; that hypholomin B can effectively enhance the phagocytosis activity of the macrophages on Escherichiacoli and kill tumor cells is confirmed. The hypholomethine B can promote the proliferation of RAW264.7 cells and promote the phagocytic activity of the RAW264.7 cells, and the hypholomethine B can promote the killing activity of the macrophages to tumor cells in a dose-dependent manner. Therefore, the new application of the hypholomin B in the aspect of immunoregulation can be used as a new indexof the effective components of phellinus linteus, and also can provide a thought for further scientific utilization of the phellinus linteus, thereby effectively improving the economic benefit of thephellinus linteus.

Description

(1) Technical field [0001] The invention relates to a new application of Hypholomine B, a medicinal monomer component of Phellinus Phellinus, in immune regulation. (2) Background technology [0002] Phellinus is a precious medicinal fungus with a history of more than 2,000 years. It is recorded in the earliest herbal book "Shen Nong's Materia Medica". It tastes slightly bitter and cold in nature. In traditional Chinese medicine, it mainly treats metrorrhagia, stranguria with blood, prolapse of the anus and diarrhea of ​​blood, leukorrhea, and amenorrhea. Modern pharmacological studies have shown that Phellinus has anti-tumor, anti-angiogenesis, anti-fibrosis, liver and liver protection, hypoglycemic, hypolipidemic, anti-oxidative, antibacterial and immune regulation effects, and is currently an internationally recognized biological cancer treatment agent A medicinal fungus with the highest efficiency. Its chemical composition is polysaccharides, fatty acids, polyphenols, t...

Claims

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Application Information

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IPC IPC(8): A61K31/366A61P37/04A61P31/04A61P35/00
CPCA61K31/366A61P31/04A61P35/00A61P37/04
Inventor 蒋福升李美芽丁滨丁志山李松涛窦晓兵吕圭源
Owner ZHEJIANG CHINESE MEDICAL UNIVERSITY
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