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Application of periplaneta americana extract and/or monomer in preparation of drugs for inhibiting M2 polarization in tumor microenvironment

A technology of Periplaneta americana and extracts, applied in the field of biomedicine, can solve problems such as not being revealed clearly, and achieve the effect of improving tumor immunoregulatory activity

Active Publication Date: 2022-03-08
DALI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Periplaneta americana extract has antibacterial, anti-inflammatory, analgesic, tissue repair and other effects. Recent studies have shown that Periplaneta americana can inhibit tumor cell proliferation and anti-tumor effects, but how to apply it and the mechanism of the effect have not been revealed clear

Method used

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  • Application of periplaneta americana extract and/or monomer in preparation of drugs for inhibiting M2 polarization in tumor microenvironment
  • Application of periplaneta americana extract and/or monomer in preparation of drugs for inhibiting M2 polarization in tumor microenvironment
  • Application of periplaneta americana extract and/or monomer in preparation of drugs for inhibiting M2 polarization in tumor microenvironment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Effect of pericanaside on the survival rate of RAW264.7 / Ana-1 cells

[0047] Experimental method: RAW264.7 cells were cultured with DMEM medium, and Ana-1 cells were cultured with 1640 medium, which contained 10% fetal bovine serum, 100 U / ml penicillin and 100 U / ml streptomycin. Cells will be at a density of 5 x 10 4 cells / mL were planted in a 96-well plate, and the blank group, control group and experimental group were set up, the old medium was discarded, 100 μL of blank medium was added to the blank group, and 100 μL of cell suspension was added to the negative group and experimental group respectively. ℃5%CO 2 After culturing in the incubator for 24 hours, discard the medium, add 100 μL medium to the blank group and the negative group, and add 100 μL to the experimental group. mL medium, 5 replicate wells per group, at 37°C 5% CO 2 Treat in the incubator for 24 hours, add 20 μL (5 mg / mL) MTT solution to each well and continue to store at 37 °C in 5% CO 2 Incubat...

Embodiment 2

[0050] Effects of pericanaside on M1 / M2 polarization markers iNOS, ARG-1, CD206

[0051] Experimental method: RT-PCR method was used to detect the expression of genes iNOS, ARG-1, and CD206, and the total RNA of RAW264.7 and Ana-1 cells was extracted by RNA-easy separation reagent method according to the manufacturer's instructions.

[0052] First-strand cDNA for qPCR was synthesized using HiScriptII QRt SuperMIX and analyzed by real-time quantitative PCR using ChamQUniversal SYBR qPCR Master Mix. The reaction mixture containing SYBR Green was composed according to the manufacturer's protocol, GAPDH was used as an internal reference, and other primers were synthesized by Bioengineering Co., Ltd. (Table 1). PCR results were analyzed by Bio-Rad CFX Manager software through 2 -ΔΔCt analyze data.

[0053] Table 1 Primer sequences for RT-PCR

[0054]

[0055] The effects of Periplaneta americana crude extract CⅡ-3 and pericanaside on iNOS, ARG-1 and CD206 as shown in figure...

Embodiment 3

[0058] Effects of Periplaneta americana extract CⅡ-3 and pericanaside on M2 markers IL-10 and TGF-β

[0059] Experimental method: RAW264.7 and Ana-1 cells were seeded in 12-well plates (10 5 cells / mL), the cells were treated according to the grouping situation, and the cell culture supernatant was collected and centrifuged. ELISA kits were used to detect M2 macrophage markers TGF-β and IL-10. A microplate reader sequentially detects the optical density (OD) value at 450 nm.

[0060] Experimental results: Crude extract of Periplaneta americana CⅡ-3 (150μg / mL, 75μg / mL, 37.5μg / mL) and pericanaside (50μg / mL, 25μg / mL, 12.5μg / mL) on IL-10, TGF-β the impact of Figure 5 and Figure 6 As shown, after IL-4 treated RAW264.7 / Ana-1 cells, the expressions of TGF-β and IL-10 were significantly increased (P<0.001). The expression levels of ARG-1 and CD206 in RAW264.7 cells decreased significantly in a dose-dependent manner (P<0.05 or P<0.001). In Ana-1 cells, high doses of CⅡ-3 and per...

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Abstract

The invention provides an application of a periplaneta americana extract and / or a periplaneta americana monomer in preparation of drugs for inhibiting M2 polarization in a tumor microenvironment, and relates to the technical field of biological medicines. According to the invention, a macrophage M1 / M2 polarization model is used as an evaluation means, and a monomeric substance perianaside which can significantly inhibit macrophage from promoting tumor M2 phenotypic polarization is separated and identified from a tumor proliferation inhibition active site CII3 of periplaneta americana. Results show that perianaside can significantly inhibit expression of M2 type macrophage markers, and it is prompted that perianaside has a very good inhibiting effect on promotion of tumor M2 type polarization. From the aspects of improving the tumor immunomodulatory activity and inhibiting M2 polarization type macrophages, the anti-tumor effect and the action mechanism of the periplaneta americana extract CII3 and the periplaneta americana extract CII3 are elaborated, and the periplaneta americana extract CII3 can be used as a tumor proliferation inhibitor and a macrophage M1 / M2 polarization inhibitor.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of Periplaneta americana extract and / or monomer in the preparation of drugs for inhibiting M2 polarization of tumor microenvironment. Background technique [0002] Cancer is one of the major diseases affecting human health, causing the death of many cancer patients around the world every year. The immune response is the body's natural barrier against tumors, and the imbalance of immune effects is an important reason for the occurrence and development of tumors. Macrophages are highly plastic innate immune cells that can perform different functions according to the stimuli of the surrounding environment. Macrophages are functionally divided into M1 phenotype and M2 phenotype. Furthermore, tumor microenvironment (TME) macrophages are activated by tumor-derived cytokines into an M2 polarized phenotype to promote tumor progression. Therefore, the de...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/64A61K31/7048A61P35/00A61P35/02
CPCA61K35/64A61K31/7048A61P35/00A61P35/02A61K2300/00Y02A50/30
Inventor 肖怀张成桂许静蕾车逸豪刘衡刘馨月杨大松刘光明
Owner DALI UNIV
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