PMID- 28778215 OWN - NLM STAT- MEDLINE DCOM- 20171016 LR - 20220316 IS - 2008-2231 (Electronic) IS - 1560-8115 (Print) IS - 1560-8115 (Linking) VI - 25 IP - 1 DP - 2017 Aug 4 TI - Pinus densiflora needle supercritical fluid extract suppresses the expression of pro-inflammatory mediators iNOS, IL-6 and IL-1beta, and activation of inflammatory STAT1 and STAT3 signaling proteins in bacterial lipopolysaccharide-challenged murine macrophages. PG - 18 LID - 10.1186/s40199-017-0184-y [doi] LID - 18 AB - BACKGROUND: Regulation of a persistently-activated inflammatory response in macrophages is an important target for treatment of various chronic diseases. Pine needle extracts are well known to have potent immunomodulatory effects. The current study was designed to evaluate the effects of Pinus densiflora needle supercritical fluid extract (PDN-SCFE) on bacterial lipopolysaccharide (LPS)-induced inflammatory response in RAW 264.7 murine macrophages. METHODS: Cytotoxic effect of PDN-SCFE was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The levels of nitric oxide (NO) and the corresponding enzyme, inducible nitric oxide synthase (iNOS), were quantified by Griess and immunoblotting methods, respectively. The levels of cytokines were quantified using commercial ELISA kits. Quantitative real-time PCR (qRT-PCR) analysis was performed to assess the mRNA expression of iNOS and cytokines. To elucidate the mechanism of action, the involvement of nuclear transcription factor-kappa B (NFkappaB), mitogen activated protein kinases (MAPKs) and Janus kinase-signal transducers and activators of transcription (JAK-STAT) pathways were examined by an immunoblotting method. In addition, the cellular localization of NFkappaB was analyzed by immunofluorescence staining. RESULTS: MTT assay results indicated that PDN-SCFE is non-toxic to RAW 264.7 cells up to a maximum assayed concentration of 40 mug/mL. The PDN-SCFE exhibited a concentration-dependent inhibitory effect on LPS-induced NO production by down regulating the expression of iNOS. In addition, the extract suppressed the LPS-induced expression of interleukin-6 (IL-6) and interleukin-1beta (IL-1beta) but not tumour necrosis factor-alpha (TNFalpha). Mechanistic studies revealed that PDN-SCFE does not influence the NFkappaB and MAPK pathways. However, it showed a significant inhibitory effect on LPS-induced activation of STAT1 and STAT3 proteins in macrophages. CONCLUSION: The present findings revealed that the anti-inflammatory activity of PDN-SCFE in LPS-challenged RAW 264.7 macrophages is probably caused by the suppression of the JAK-STAT signaling pathway. FAU - Venkatesan, Thamizhiniyan AU - Venkatesan T AD - Department of Forest Products and Biotechnology, College of Forest Science, Kookmin University, 861-1 Chongnung-dong, Songbuk-gu, Seoul, 136-702, South Korea. FAU - Choi, Young-Woong AU - Choi YW AD - Department of Forest Products and Biotechnology, College of Forest Science, Kookmin University, 861-1 Chongnung-dong, Songbuk-gu, Seoul, 136-702, South Korea. FAU - Lee, Jennifer AU - Lee J AD - Department of Forest Products and Biotechnology, College of Forest Science, Kookmin University, 861-1 Chongnung-dong, Songbuk-gu, Seoul, 136-702, South Korea. FAU - Kim, Young-Kyoon AU - Kim YK AD - Department of Forest Products and Biotechnology, College of Forest Science, Kookmin University, 861-1 Chongnung-dong, Songbuk-gu, Seoul, 136-702, South Korea. ykkim@kookmin.ac.kr. LA - eng PT - Journal Article DEP - 20170804 PL - Switzerland TA - Daru JT - Daru : journal of Faculty of Pharmacy, Tehran University of Medical Sciences JID - 101125969 RN - 0 (Interleukin-18) RN - 0 (Interleukin-6) RN - 0 (Lipopolysaccharides) RN - 0 (Plant Extracts) RN - 0 (STAT1 Transcription Factor) RN - 0 (STAT3 Transcription Factor) RN - 0 (Stat1 protein, mouse) RN - 0 (Stat3 protein, mouse) RN - EC 1.14.13.39 (Nitric Oxide Synthase Type II) SB - IM MH - Animals MH - Blotting, Western MH - Inflammation/drug therapy MH - Interleukin-18/*antagonists & inhibitors MH - Interleukin-6/*antagonists & inhibitors MH - Lipopolysaccharides/pharmacology MH - Macrophages/*drug effects MH - Mice MH - Microscopy, Fluorescence MH - Nitric Oxide Synthase Type II/*antagonists & inhibitors MH - Pinus/*chemistry MH - Plant Extracts/*pharmacology MH - Plant Leaves/*chemistry MH - RAW 264.7 Cells MH - Real-Time Polymerase Chain Reaction MH - STAT1 Transcription Factor/*antagonists & inhibitors MH - STAT3 Transcription Factor/*antagonists & inhibitors PMC - PMC5544993 OTO - NOTNLM OT - Pine needle OT - Pinus densiflora OT - RAW 264.7 murine macrophages OT - Supercritical fluid extract COIS- ETHICS APPROVAL AND CONSENT TO PARTICIPATE: Not applicable. CONSENT FOR PUBLICATION: Not applicable. COMPETING INTERESTS: The authors declare that they have no competing interests. PUBLISHER'S NOTE: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. EDAT- 2017/08/06 06:00 MHDA- 2017/10/17 06:00 PMCR- 2017/08/04 CRDT- 2017/08/06 06:00 PHST- 2017/03/30 00:00 [received] PHST- 2017/07/27 00:00 [accepted] PHST- 2017/08/06 06:00 [entrez] PHST- 2017/08/06 06:00 [pubmed] PHST- 2017/10/17 06:00 [medline] PHST- 2017/08/04 00:00 [pmc-release] AID - 10.1186/s40199-017-0184-y [pii] AID - 184 [pii] AID - 10.1186/s40199-017-0184-y [doi] PST - epublish SO - Daru. 2017 Aug 4;25(1):18. doi: 10.1186/s40199-017-0184-y.