Inhibitory Effect and Mechanism of Jingulian Extract on LPS-induced RAW264.7 Cell Inflammatory Response Based on PI3K/Akt Signaling Pathway
10.13422/j.cnki.syfjx.20211401
- VernacularTitle:基于PI3K/Akt信号通路探讨金骨莲提取物抑制脂多糖诱导RAW264.7细胞炎症反应的作用及机制
- Author:
Si-han LI
1
;
Dong-yin LIAN
2
;
Guang-ping ZHANG
2
;
Ying CHEN
2
;
Jian-rong LI
2
;
Zu-guang YE
2
;
Bo PENG
2
Author Information
1. Guizhou University，Guiyang 550025，China
2. Institute of Chinese Materia Medica， China Academy of Chinese Medical Sciences，Beijing 100700，China
- Publication Type:Research Article
- Keywords:
Jingulian extract （JGL）;
lipopolysaccharide （LPS）;
RAW264.7 cell;
inflammation;
phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(14):29-35
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the inhibitory effect and mechanism of Jingulian extract （JGL） on inflammation. Method:The following groups were set up in this study： a control group （10% fetal bovine serum）， a lipopolysaccharide （LPS） model group （0.5 mg·L-1）， and JGL groups （10， 20， 40， 60， 80， 120， 160， 200， 250， 300 mg·L-1 + 0.5 mg·L-1 LPS）. The RAW264.7 cells were cultured for 24 hours. Cell proliferation was detected by cell counting kit-8 （CCK-8） assay. Nitric oxide （NO） release was detected by Griess assay. The release of cytokines interleukin （IL）-1β， IL-6， IL-10， and tumor necrosis factor （TNF）-α was determined by enzyme linked immunosorbent assay （ELISA）. The expression of inducible nitric oxide synthase （iNOS） and intraprostaglandin peroxidase synthase 2 （PTGS2）/cyclooxygenase-2 （COX-2） was measured by real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） and the activation of key proteins in the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway by Western blot. Result:Compared with the control group， LPS （0.5 mg·L-1）could promote the proliferation of RAW264.7 cells after stimulation for 24 hours （P<0.01）. Compared with the model group， JGL had no significant effect on cell proliferation. Compared with the control group， LPS （0.5 mg·L-1）increased the release of NO， IL-1β， IL-6， IL-10， and TNF-α （P<0.01）. Compared with the model group， JGL （20-300 mg·L-1）inhibited the release of NO in a dose-dependent manner after stimulation for 24 hours （P<0.05） and reduced IL-1β， IL-6， and IL-10 （P<0.05， P<0.01）， but no obvious inhibition on the release of TNF-α was observed. LPS （0.5 mg·L-1） could induce the expression of iNOS and PTGS2/COX-2 genes as compared with the control group （P<0.05， P<0.01）. JGL could down-regulate the mRNA expression of iNOS and PTGS2/COX-2 genes as compared with the model group （P<0.05， P<0.01）. LPS （0.5 mg·L-1） could activate the PI3K/Akt pathway （P<0.01） as compared with the control group， while JGL （10， 20， 40， and 80 mg·L-1） decreased the expression of PI3K-p110， p-p85， and p-Akt （P<0.01）， and inhibited the activation of PI3K/Akt pathway. Conclusion:JGL extract could significantly inhibit the inflammatory response and activation of the PI3K/Akt pathway induced by LPS in RAW264.7 cells. The anti-inflammatory effect was related to the inhibition of the PI3K/Akt pathway.
