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Glycyrrhetinic acid-induced hepatic cell oxidative stress and inflammatory response and application thereof

A technology of glycyrrhetic acid and hepatocytes, applied in the field of biomedicine, can solve problems such as glycyrrhetic acid that have not yet been seen

Active Publication Date: 2022-05-13
INST OF ENVIRONMENTAL MEDICINE & OCCUPATIONAL MEDICINE ACAD OF MILITARY MEDICINE ACAD OF MILITARY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The protective effect of GA on the liver has been widely reported in various liver diseases, but so far, there has been no relevant report on the effect of glycyrrhetinic acid on normal liver cells

Method used

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  • Glycyrrhetinic acid-induced hepatic cell oxidative stress and inflammatory response and application thereof
  • Glycyrrhetinic acid-induced hepatic cell oxidative stress and inflammatory response and application thereof
  • Glycyrrhetinic acid-induced hepatic cell oxidative stress and inflammatory response and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Effect of glycyrrhetinic acid on human normal liver cell L-02 cell viability

[0046] 1. Experimental materials

[0047] The structural formulas of glycyrrhizic acid, glycyrrhetinic acid, and liquiritin are as follows figure 1 As shown in A, all were purchased from Shanghai Yuanye Biotechnology Co., Ltd.; human normal liver cell L-02 cell line was purchased from Peking Union Medical College Cell Bank, and the antibodies used for Western blot included antibodies against NF-κB p105 / p50 (Cell Signaling Technology, USA, #3035, 1:1200), NF-κB p100 / p52 (Cell Signaling Technology, USA, #4882, 1:1500), TNF receptor-related factor 1TRAF1 (Abcam, UK, ab155268, 1:1200) , TRAF3 (Abcam, UK, ab155298, 1:800), CXCL8 (Abcam, UK, ab18672, 1:2000), CyclinD1 (Abcam, ab226977, 1:1000), Survivin (Bioss, bs-0615R, 1:500) , MMP9 (Abcam, ab58803, 1:1500), TRIM25 (Bioss, bs-4170R, 1:500) and GAPDH (UtiBody, UM4002, 1:2000) antibodies.

[0048] 2. Cell culture and glycyrrhetinic aci...

Embodiment 2

[0054] Example 2 Glycyrrhetinic acid induces oxidative stress in human normal liver cells L-02

[0055] 1. Experimental method

[0056] The oxidative stress of L-02 cells treated with different concentrations (25 μM, 50 μM, 100 μM) of glycyrrhetinic acid or DMSO for 48 h was detected by commercially available kits (Nanjing Jiancheng Institute of Bioengineering, Nanjing, China).

[0057] (1) Detection of intracellular reactive oxygen species (ROS) and superoxide dismutase (SOD) levels

[0058] The level of reactive oxygen species (ROS) was measured by incubating L-02 cells with 2,7-dichlorofluorescein diacetate (DCFH-DA) at 37°C for 1 h. L-02 cells were cultured in a 6-well plate, supplemented with 10 μM / L DCFH-DA, and incubated at 37°C for 20 minutes. Cells were then washed 3 times with serum-free medium. L-02 cells were treated with 0, 25, 50 and 100 μM glycyrrhetinic acid, and the cells were collected after treatment, and then the fluorescent signal was detected by a fluo...

Embodiment 3

[0066] Example 3 Effect of Glycyrrhetinic Acid on Transcription Regulation of Human Normal Hepatocyte L-02

[0067] 1. RNA sequencing

[0068] In order to fully elucidate the effect of glycyrrhetinic acid on the gene expression of L-02 cells, this example uses the Trizol kit (Promega, Madison, USA) to extract the total RNA of L-02 cells, regardless of whether it has been treated with glycyrrhetinic acid or not. its total RNA.

[0069] L-02 cells were cultured in 6-well plates (1 × 10 per well 6 cells), and then treated L-02 cells with 0 μM, 25 μM, 50 μM, 100 μM glycyrrhetinic acid. Cells were washed with PBS and collected for subsequent total RNA extraction. Total RNA was extracted according to the instructions. The quality of RNA in the extracts was verified using an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). The RNA was reverse-transcribed into cDNA, which was then purified using a commercially available kit (QIAquick PCR Extraction Kit, Prom...

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Abstract

The invention discloses glycyrrhetinic acid induced hepatic cell oxidative stress and inflammatory response and application thereof, and finds and verifies that glycyrrhetinic acid has hepatotoxicity and can cause liver injury for the first time. The invention provides an experimental cell line model and an animal model which are used for drug liver injury research and are similar to the pathological mechanism of human liver diseases for the field, an experimental basis is provided for further exploration of the mechanism of glycyrrhetinic acid to generate hepatotoxicity, and a new thought is provided for screening and development of novel drugs aiming at hepatotoxicity.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. Specifically, the invention relates to glycyrrhetinic acid inducing liver cell oxidative stress and inflammatory response and application thereof. Background technique [0002] Glycyrrhiza uralensis Fisch, a leguminous plant, has been widely reported for its antioxidant and immunomodulatory effects, and it contains many bioactive components, such as glycyrrhizic acid (GL), glycyrrhetinic acid ( Glycyrrhetinic acid, GA), Liquiritin, etc. Among them, GA is an important effective active ingredient extracted from the rhizome of licorice, which belongs to pentacyclic triterpene saponins, and is similar in structure to steroid hormones. At present, glycyrrhetinic acid and its derivatives have been proved to have a variety of pharmacological effects, including anti-inflammatory, antibacterial, antiviral, anti-tumor and antioxidant activities, etc. Glycyrrhetinic acid and its derivatives can inhibit...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A01K67/027A61P1/16A61K31/56C12R1/91
CPCC12N5/067A01K67/027A61K31/56A61P1/16C12N2500/30C12N2500/02A01K2207/20A01K2267/03A01K2267/0368Y02A50/30
Inventor 吴瑾王永辉李晓丽刘忠文高月王宇光席令仪
Owner INST OF ENVIRONMENTAL MEDICINE & OCCUPATIONAL MEDICINE ACAD OF MILITARY MEDICINE ACAD OF MILITARY SCI
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