Xiaojindan Extract Modulated Macrophage Polarization by Targeting PI3K/Akt Pathway
10.13422/j.cnki.syfjx.20220724
- VernacularTitle:基于PI3K/Akt信号通路探讨小金丹对巨噬细胞极化的调控作用及机制
- Author:
Bo PENG
1
;
Dong-yin LIAN
1
;
Guang-ping ZHANG
1
;
Ying CHEN
1
;
Hong-ping HOU
1
;
Rong HE
1
;
Jian-rong LI
1
;
Xiao-ru HU
2
Author Information
1. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China
2. National Institutes for Food and Drug Control, Beijing 100050, China
- Publication Type:Journal Article
- Keywords:
Xiaojindan;
RAW264.7;
macrophage polarization;
phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2022;28(9):36-42
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo explore the effect and mechanism of Xiaojindan extract (XJD) on macrophage polarization. MethodLipopolysaccharide (LPS) and interleukin-4 (IL-4) were used to induce M1 and M2 polarization of RAW264.7 cells. The influence of 10-80 mg·L-1 XJD on cell proliferation was detected by Cell Counting Kit-8 (CCK-8) assay. Nitric oxide (NO) and interleukin-6 (IL-6) release was explored by Griess assay and enzyme-linked immunosorbent assay (ELISA), respectively. The mRNA expression of M1 and M2 macrophage markers was measured by real-time quantitative polymerase chain reaction (Real-time PCR), and the CD206+ expression was determined by flow cytometry. The activation of phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathway was analyzed by western blot. Result10-80 mg·L-1 XJD showed no marked cytotoxicity in LPS (0.5 mg·L-1)- or IL-4 (20 μg·L-1)-induced RAW264.7 cells. Compared with the control group, LPS significantly promoted the expression of M1 macrophage markers (P<0.01), including increased NO and IL-6 release (P<0.01) and upregulated mRNA expression of interleukin-1β (IL-1β), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and tumor necrosis factor-α (TNF-α) (P<0.01). Compared with LPS-induced group, 20-80 mg·L-1 XJD decreased the release of NO and IL-6 in a dose-dependent manner (P<0.01), and similarly 10-80 mg·L-1 XJD suppressed the mRNA expression of IL-1β, iNOS, COX-2 and TNF-α (P<0.01). Compared with the control group, IL-4 obviously increased the expression of M2 macrophage markers (P<0.01), including increased CD206+ cell population and upregulated mRNA expression of arginine-1 (Arg-1), interleukin-10 (IL-10), interleukin-13 (IL-13) and transforming growth factor-β1 (TGF-β1). Compared with IL-4-induced group, 10-80 mg·L-1 XJD dose-dependently decreased CD206+ cell population (P<0.01) and inhibited the mRNA expression of Arg-1, IL-10, IL-13 and TGF-β1 (P<0.01). Western blot showed that XJD significantly downregulated the activation of PI3K/Akt pathway as compared to LPS- and IL-4-induced groups (P<0.05, P<0.01). ConclusionXJD significantly inhibited the macrophage polarization in the LPS- and IL-4-induced RAW264.7 cells by targeting PI3K/Akt pathway.