Study on anti-inflammatory active components and mechanism of Corydalis Bungeanae Herba

LI Mu-zi1,2 ZHANG Hao1,2 CUI Guo-qian1,2 XIANG Yan1,2 ZHU Fen-xia1,2

(1.Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, China 210028)
(2.Key Laboratory of New Drug Delivery System of Chinese Materia Medica, Jiangsu Academy of Traditional Chinese Medicine, Nanjing, China 210028)

【Abstract】Corydalis Bungeanae Herba is often used to treat a variety of inflammatory diseases in traditional Chinese medicine. In order to determine its chemical material basis, the components of Corydalis Bungeanae Herba were isolated by an automated purification system. Flavonoids and alkaloids were prepared, and all such components were identified by mass spectrometry. The effects of the components on the production of inflammatory mediators and pharmacological mechanisms in the lipopolysaccharide (LPS)-induced RAW264.7 cell inflammation model were examined. Mouse macrophages (RAW264.7) were first treated with LPS. The relationship between cell viability and LPS concentration was observed. Then, the effects of flavonoid and alkaloid components with different administration concentrations on cell viability were detected to determine the maximum administration concentration. Secondly, 2.5, 5, 10 and 20 μg·mL−1 flavonoid and alkaloid components were added respectively to observe the effects and mechanism of different concentrations of flavonoid and alkaloid components on LPS-induced inflammation of RAW264.7 macrophages. Griess reagent assay was used to detect NO content in cell supernatant. The inflammatory cytokines (TNF-α, IL-1β and IL-6) in cell supernatant were determined by ELISA method. Western blot method was used to detect the intracellular nuclear factor (NF-κB) IκBα phosphorylation (p-IκBα), p65 phosphorylation (p-p65) and protein expression of TLR4 and TLR2. The results showed that the alkaloid components inhibited the production of NO, TNF-α, IL-1β and IL-6 in a dose-dependent manner in the concentration range of 2.5–20 μg·mL−1. In inflammation upstream pathways, the inhibitory effect of the alkaloid components on the TLR2 expression level was weaker than that of TLR4. In inflammation downstream, alkaloid components significantly inhibited the phosphorylation of IκBα and p65 in a dose-dependent manner. These data suggested that the alkaloid components were the material basis of Corydalis Bungeanae Herba, and its anti-inflammatory mechanism might be related to inhibiting the transmission of inflammatory signals in TLRs/NF-κB signaling pathways dominated by TLR4, interfering with the activation of inflammatory genes and inhibiting their over expression, and down-regulating the secretion level of inflammatory factors.

【Keywords】 Corydalis Bungeanae Herba; inflammation; alkaloids; flavonoids; RAW264.7 cell;

【DOI】

【Funds】 National Natural Science Foundation of China (81473430) Jiangsu Medical Innovation Team Project (CXTDB2017003)

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(Translated by FU LJ)

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This Article

ISSN:1001-5302

CN:11-2272/R

Vol 45, No. 11, Pages 2586-2594

June 2020

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Article Outline

Abstract

  • 1 Materials
  • 2 Methods
  • 3 Results
  • 4 Discussion
  • References