Anti-inflammatory effects of ethyl acetate fraction from Melilotus suaveolens Ledeb on LPS-stimulated RAW 264.7 cells.
OBJECTIVE
This paper aimed to elucidate the anti-inflammatory effects of EtOAc fraction prepared from Melilotus suaveolens Ledeb ethanol extract with a cellular model of LPS-stimulated RAW 264.7 cell.


METHODS
Some key pro-inflammatory cytokines and mediators including IL-1 beta, IL-6, NO, iNOS, COX-2 and TNF-alpha, two important anti-inflammatory cytokines and mediators IL-10 and HO-1, I-kappaB and NF-kappaB were studied by sandwich ELISA, real-time PCR, western blot analysis and immunocytochemistry. At last a HPLC fingerprint was taken to evaluate the fraction.


RESULTS
The EtOAc fraction could significantly inhibit the production of IL-1 beta, IL-6, NO, TNF-alpha, COX-2 in LPS-stimulated cell than that of single LPS-stimulated cell (p<0.01 or p<0.05), and the extract could increase the production of IL-10 and HO-1 than that of single LPS intervention cell (p<0.01 or p<0.05). Meanwhile, the extract also could inhibit the production of NF-kappaB compared to single LPS-stimulated cell. All the results showed that the extract had a good anti-inflammatory effect on LPS-stimulated RAW264.7 cell.


CONCLUSIONS
Taken together, the anti-inflammatory actions of M. suaveolens Ledeb EtOAc fraction might be due to the down-regulation of IL-1 beta, IL-6, NO, TNF-alpha and COX-2 via the suppression of NF-kappaB activation, and another pathway was up regulating the production of IL-10 and HO-1. Meanwhile, the EtOAc fraction might be further studied to isolate the active anti-inflammatory ingredients besides coumarin.
State Key Laboratory of Virology, College of Life Sciences, Chinese-French Liver Disease Research Institute of Wuhan University at Zhongnan Hospital, Wuhan University, Wuhan, 430072, PR China.
Jun-Yan
JY
Tao
Guo-Hua
GH
Zheng
Lei
L
Zhao
Jian-Guo
JG
Wu
Xiao-Yu
XY
Zhang
Shu-Ling
SL
Zhang
Zhi-Jun
ZJ
Huang
Fu-Liang
FL
Xiong
Chong-Ming
CM
Li
D000085:Acetates / Q000737:chemistry*; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000627:therapeutic use*; D001483:Base Sequence; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D016207:Cytokines / Q000032:analysis; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D018836:Inflammation Mediators / Q000032:analysis; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008262:Macrophage Activation; D008264:Macrophages / Q000187:drug effects; D029903:Melilotus / Q000737:chemistry*; D051379:Mice; D010936:Plant Extracts / Q000627:therapeutic use*; D016133:Polymerase Chain Reaction
D000085:Acetates; D000893:Anti-Inflammatory Agents; D016207:Cytokines; D017931:DNA Primers; D018836:Inflammation Mediators; D008070:Lipopolysaccharides; D010936:Plant Extracts; C007650:ethyl acetate