Isolation and characterization of LHT-type plant amino acid transporter gene from Panax ginseng Meyer

Zhang, Ru; Zhu, Jie; Cao, Hongzhe; Xie, Xiaolei; Huang, Jingjia; Chen, Xianghui; Luo, Zhitian

doi:10.5142/jgr.2013.37.361

Cited by 22 publications

(14 citation statements)

References 27 publications

(38 reference statements)

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“…Total RNA was isolated with TRIzol Reagent (Gibco BRL) according to the manufacturer's instructions and stored at −70°C until use. The mRNA was quantified by real-time reverse transcriptase polymerase chain reaction (RT-PCR) with SYBR Premix Ex Taq, according to the manufacturer's instructions (Takara, Shiga, Japan), using a real-time thermal cycler (Bio-Rad, Hercules, CA, USA), as reported previously [23,24] . Results were expressed as the ratio of the optical density relative to glyceraldehyde 3-phosphate dehydrogenase.…”

Section: Methodsmentioning

confidence: 99%

Molecular mechanism of protopanaxadiol saponin fraction-mediated anti-inflammatory actions

Yang

Lee

Rhee

et al. 2015

Journal of Ginseng Research

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BackgroundKorean Red Ginseng (KRG) is a representative traditional herbal medicine with many different pharmacological properties including anticancer, anti-atherosclerosis, anti-diabetes, and anti-inflammatory activities. Only a few studies have explored the molecular mechanism of KRG-mediated anti-inflammatory activity.MethodsWe investigated the anti-inflammatory mechanisms of the protopanaxadiol saponin fraction (PPD-SF) of KRG using in vitro and in vivo inflammatory models.ResultsPPD-SF dose-dependently diminished the release of inflammatory mediators [nitric oxide (NO), tumor necrosis factor-α, and prostaglandin E2], and downregulated the mRNA expression of their corresponding genes (inducible NO synthase, tumor necrosis factor-α, and cyclooxygenase-2), without altering cell viability. The PPD-SF-mediated suppression of these events appeared to be regulated by a blockade of p38, c-Jun N-terminal kinase (JNK), and TANK (TRAF family member-associated NF-kappa-B activator)-binding kinase 1 (TBK1), which are linked to the activation of activating transcription factor 2 (ATF2) and interferon regulatory transcription factor 3 (IRF3). Moreover, this fraction also ameliorated HCl/ethanol/-induced gastritis via suppression of phospho-JNK2 levels.ConclusionThese results strongly suggest that the anti-inflammatory action of PPD-SF could be mediated by a reduction in the activation of p38-, JNK2-, and TANK-binding-kinase-1-linked pathways and their corresponding transcription factors (ATF2 and IRF3).

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Section: Methodsmentioning

confidence: 99%

Molecular mechanism of protopanaxadiol saponin fraction-mediated anti-inflammatory actions

Yang

Lee

Rhee

et al. 2015

Journal of Ginseng Research

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“…Total RNA was stored at −70°C until use. Semiquantitative RT reactions were conducted as previously reported [ 39 , 40 ]. Quantification of mRNA was performed by real-time RT-PCR with SYBR Premix Ex Taq according to the manufacturer's instructions (Takara, Shiga, Japan) using a real-time thermal cycler (Bio-Rad, Hercules, CA), as reported previously [ 41 ].…”

Section: Methodsmentioning

confidence: 99%

The Dietary Flavonoid Kaempferol Mediates Anti‐Inflammatory Responses via the Src, Syk, IRAK1, and IRAK4 Molecular Targets

Kim

Park

Lee

et al. 2015

Mediators of Inflammation

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Even though a lot of reports have suggested the anti-inflammatory activity of kaempferol (KF) in macrophages, little is known about its exact anti-inflammatory mode of action and its immunopharmacological target molecules. In this study, we explored anti-inflammatory activity of KF in LPS-treated macrophages. In particular, molecular targets for KF action were identified by using biochemical and molecular biological analyses. KF suppressed the release of nitric oxide (NO) and prostaglandin E2 (PGE2), downregulated the cellular adhesion of U937 cells to fibronectin (FN), neutralized the generation of radicals, and diminished mRNA expression levels of inflammatory genes encoding inducible NO synthase (iNOS), TNF-α, and cyclooxygenase- (COX-) 2 in lipopolysaccharide- (LPS-) and sodium nitroprusside- (SNP-) treated RAW264.7 cells and peritoneal macrophages. KF reduced NF-κB (p65 and p50) and AP-1 (c-Jun and c-Fos) levels in the nucleus and their transcriptional activity. Interestingly, it was found that Src, Syk, IRAK1, and IRAK4 responsible for NF-κB and AP-1 activation were identified as the direct molecular targets of KF by kinase enzyme assays and by measuring their phosphorylation patterns. KF was revealed to have in vitro and in vivo anti-inflammatory activity by the direct suppression of Src, Syk, IRAK1, and IRAK4, involved in the activation of NF-κB and AP-1.

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“…All RNA was stored at -70℃ until use. Semi-quantitative RT reactions were conducted as previously reported [ 20 21 ]. mRNA quantification was performed by real-time RT-PCR with SYBR Premix Ex Taq according to the manufacturer's instructions (Takara, Shiga, Japan) using a real-time thermal cycler (Bio-Rad, Hercules, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

Scutellarein Reduces Inflammatory Responses by Inhibiting Src Kinase Activity

Sung

Kim

Cho

2015

Korean J Physiol Pharmacol

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Flavonoids are plant pigments that have been demonstrated to exert various pharmacological effects including anti-cancer, anti-diabetic, anti-atherosclerotic, anti-bacterial, and anti-inflammatory activities. However, the molecular mechanisms in terms of exact target proteins of flavonoids are not fully elucidated yet. In this study, we aimed to evaluate the anti-inflammatory mechanism of scutellarein (SCT), a flavonoid isolated from Erigeron breviscapus, Clerodendrum phlomidis and Oroxylum indicum Vent that have been traditionally used to treat various inflammatory diseases in China and Brazil. For this purpose, a nitric oxide (NO) assay, polymerase chain reaction (PCR), nuclear fractionation, immunoblot analysis, a kinase assay, and an overexpression strategy were employed. Scutellarein significantly inhibited NO production in a dose-dependent manner and reduced the mRNA expression levels of inducible NO synthase (iNOS) and tumor necrosis factor (TNF)-α in lipopolysaccharide (LPS)-activated RAW264.7 cells. In addition, SCT also dampened nuclear factor (NF)-κB-driven expression of a luciferase reporter gene upon transfection of a TIR-domain-containing adapter-inducing interferon-β (TRIF) construct into Human embryonic kidney 293 (HEK 293) cells; similarly, NF-κ B nuclear translocation was inhibited by SCT. Moreover, the phosphorylation levels of various upstream signaling enzymes involved in NF-κB activation were decreased by SCT treatment in LPS-treated RAW264.7 cells. Finally, SCT strongly inhibited Src kinase activity and also inhibited the autophosphorylation of overexpressed Src. Therefore, our data suggest that SCT can block the inflammatory response by directly inhibiting Src kinase activity linked to NF-κB activation.

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Isolation and characterization of LHT-type plant amino acid transporter gene from Panax ginseng Meyer

Cited by 22 publications

References 27 publications

Molecular mechanism of protopanaxadiol saponin fraction-mediated anti-inflammatory actions

Molecular mechanism of protopanaxadiol saponin fraction-mediated anti-inflammatory actions

The Dietary Flavonoid Kaempferol Mediates Anti‐Inflammatory Responses via the Src, Syk, IRAK1, and IRAK4 Molecular Targets

Scutellarein Reduces Inflammatory Responses by Inhibiting Src Kinase Activity

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