• Journal of Life Science
• /
• v.33 no.2
• /
• pp.176-182
• /
• 2023

The stems of Dendrobium moniliforme are used in traditional Oriental medicine as a Yin tonic to nourish the stomach, promote the production of body fluid, and reduce fever. This study investigated the effects of the aqueous extract of D. moniliforme stems (DME) on mast cell degranulation and the expression of tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4), and histamine-synthesizing enzyme histidine decarboxylase (HDC). We used rat mast cell line RBL-2H3 cells and stimulated them with PMA plus calcium ionophore (PMACI). Pretreatment with DME significantly inhibited PMACI-induced β-hexosaminidase release and the expression of TNF-α, IL-4, and HDC. Furthermore, DME suppressed PMACI-induced nuclear translocation of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and activator protein 1 (AP-1). In addition, HDC expression was inhibited by SP600125 (JNK inhibitor), PD98059 (ERK inhibitor), and SB203580 (p38 kinase inhibitor). Finally, the phosphorylation of p38 kinase, extracellular signal-regulated kinase 1/2 (ERK1/2), and c-Jun N-terminal kinase (JNK) was inhibited by pretreatment with DME. These results suggest that DME has inhibitory effects against degranulation, cytokine (TNF-α and IL-4) and HDC expression, and that HDC expression is mediated by MAPK signaling. These findings suggest that DME may have therapeutic potential in the treatment of hypersensitive and inflammatory diseases.

• Journal of Applied Biological Chemistry
• /
• v.66
• /
• pp.7-14
• /
• 2023

The effects of fermented Achyranthes japonica Nakai, Angelica gigas Nakai, and Eucommia ulmoides Oliver extracts (FAAE) on regulation of inflammation and apoptosis were investigated in primary cultured rat cartilage cells. To identify the protective effects of FAAE against H₂O₂, cell survival was measured by MTT assay. Smad3, Collagen type I, MMP3, and MMP13 were measured by real-timpe PCR and westernbot and the inflammatory (NF-κB pathway, COX-2, iNOS) factors were determined by western blot. The apoptosis related factors (JNK, c-Fos, c-Jun, caspase 3, Bax, and Bcl-2) were determined by western blot. FAAE significantly increased the follwing: H₂O₂ treated cell survival, mRNA and protein expression of Smad 3, collagen type I. In addition, FAAE significantly decreased the protein expression of inflammatory and apoptosis related factors. This study suggests that FAAE have a protection effect of chondrocytes through inhibition of inflammation and apoptosis. Thus, FAAE is a therapeutic potential food componet in osteoarthritis.

• Korean Journal of Plant Resources
• /
• v.34 no.3
• /
• pp.203-215
• /
• 2021

Bok choy is one of Brassica vegetables widely consumed worldwide. Brassica vegetables have been reported to exert various pharmacological activities such as antioxidant, anti-cancer and cardioprotective activity. However, studies on immunostimulatory activity of bok choy sprout have not been conducted properly. Thus, in this study, we investigated in vitro immunostimulatory activity of bok choy sprout extract (BCS) using mouse macrophage RAW264.7 cells. Our results showed that BCS increased the production of immunomodulators such as NO, iNOS, IL-1β, IL-6, IL-12, TNF-α and MCP-1, and phagocytic activity in RAW264.7 cells. BCS activated MAPK, NF-κB and PI3K/AKT signaling pathways. However, BCS-mediated production of immunomodulators was dependent on JNK, NF-κB and PI3K/AKT signaling pathways. the mRNA expression of TLR2 were significantly increased by BCS, TLR2 inhibition by anti-TLR2 dramatically suppressed the production of immunomodulators by BCS. In addition, TLR2 inhibition by anti-TLR2 significantly reduced BCS-mediated phosphorylation level of AKT, JNK and NF-κB. From these results, BCS may have immunostimulatory activity via TLR2-MAPK, NF-κB and PI3K/AKT signaling pathways. Therefore, BCS expected to be used as a potential immune-enhancing agent.

• Clinical and Experimental Pediatrics
• /
• v.52 no.6
• /
• pp.696-700
• /
• 2009

Purpose : We screened more than 350 compounds with an endoperoxide ring structure in search of an anti-leukemic drug and found that compound 127 (c-127) could induce significant cytotoxicity in HL-60 cells. In this study, we investigated the molecular mechanisms of compound 127-induced antitumor activity on HL-60 cells. Methods : HL-60 cells were cultured in Rosewell Park Memorial Institute 1640 and cell viability was measured by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide], a tetrazole assay. Apoptosis was assessed by a DNA fragmentation test. Apoptotic machineries were determined by Western blot analysis. Results : C-127 could induce a cytotoxic effect at 24 h and apoptosis at 6 h, which was demonstrated with MTT assay and DNA fragmentation test, respectively. The apoptotic effect of this drug was caused by the activation of the intracellular caspase-8,3 activation, the cleavage of pro-apoptotic Bid, and the increase of c-Jun expression accompanied with JNK (Jun N-terminal kinases) phosphorylation. On the contrary, it increased the expression of anti-apoptotic Bcl-2 levels, leading to the induction of the induction of anti-apoptotic effect. Taken together, the present study demonstrated that c-127 was a potent inducer of cytotoxicity on HL-60 cells through apoptotic mechanisms, which included the activation of caspase family, the regulation of Bcl-2 family, and the activation of JNK signaling pathway. Conclusion : Our results suggest that c-127 has a strong antitumor activity through the regulation of various apoptotic machineries on HL-60 cells. The compound may be utilized as an effective and potentially therapeutic drug in leukemia.

• The Korean Journal of Physiology and Pharmacology
• /
• v.17 no.2
• /
• pp.139-147
• /
• 2013

Lysolipids such as LPA, S1P and SPC have diverse biological activities including cell proliferation, differentiation, and migration. We investigated signaling pathways of LPA-induced contraction in feline esophageal smooth muscle cells. We used freshly isolated smooth muscle cells and permeabilized cells from cat esophagus to measure the length of cells. Maximal contraction occurred at $10^{-6}M$ and the response peaked at 30s. To identify LPA receptor subtypes in cells, western blot analysis was performed with antibodies to LPA receptor subtypes. LPA1 and LPA3 receptor were detected at 50 kDa and 44 kDa. LPA-induced contraction was almost completely blocked by LPA receptor (1/3) antagonist KI16425. Pertussis toxin (PTX) inhibited the contraction induced by LPA, suggesting that the contraction is mediated by a PTX-sensitive G protein. Phospholipase C (PLC) inhibitors U73122 and neomycin, and protein kinase C (PKC) inhibitor GF109203X also reduced the contraction. The PKC-mediated contraction may be isozyme-specific since only $PKC{\varepsilon}$ antibody inhibited the contraction. MEK inhibitor PD98059 and JNK inhibitor SP600125 blocked the contraction. However, there is no synergistic effect of PKC and MAPK on the LPA-induced contraction. In addition, RhoA inhibitor C3 exoenzyme and ROCK inhibitor Y27632 significantly, but not completely, reduced the contraction. The present study demonstrated that LPA-induced contraction seems to be mediated by LPA receptors (1/3), coupled to PTX-sensitive G protein, resulting in activation of PLC, PKC-${\varepsilon}$ pathway, which subsequently mediates activation of ERK and JNK. The data also suggest that RhoA/ROCK are involved in the LPA-induced contraction.

• Biomedical Science Letters
• /
• v.12 no.3
• /
• pp.249-254
• /
• 2006

It has been reported that liver is a very important organ to xenotransplantation. Pig is known to be a most suitable species in transplantation of human organs. However, the physiological function of pig hepatocytes is not clear elucidated. Epidermal growth factor (EGF) is known to be a mitogen in various cell systems. Thus, we examined the effect of EGF on cell proliferation and its related signal cascades in primary cultured pig hepatocytes. EGF stimulates cell proliferation in a dose (>1ng/ml) dependent manner. EGF-induced increase of $[^3H]-thymidine$ incorporation was blocked by AG 1478 ($10^{-6}M$, an EGF receptor antagonist) genistein and herbymycin A (tyrosine kinase inhibitors, $10^{-6}M$), suggesting the role of activation and tyrosine phosphorylation of EGF receptor. In addition, EGF-induced increase of $[^3H]-thymidine$ incorporation was prevented by neomycin $(10^{-4}M)$, U73122 $(10^{-5}M)$ (phospholipase C [PLC] inhibitors), staurosporine ($(10^{-8}M)$, or bisindolylmaleimide I $(10^{-6}M)$ (protein kinase C [PKC] inhibitors), suggesting the role of PLC and PKC. Moreover, EGF-induced increase of $[^3H]-thymidine$ incorporation was blocked by PD 98059 (a p44/42 mitogen activated protein kinase [MAPK] inhibitor), SB 203580 (a p38 MAPK inhibitor), and SP 600125 (a JNK inhibitor). EGF increased the translocation of PKC from cytosol to membrane fraction and activated p42/44 MAPK, p38 MAPK and JNK. In conclusion, EGF stimulates cell proliferation via PKC and MAPK in cultured pig hepatocytes.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.30 no.3
• /
• pp.150-156
• /
• 2016

Extract of Broussometia kazinoki Rhizodermatis has been traditionally used for geopoong, diuresis, hwalhyeol. In the present study, the apoptotic effect of methanol extract of Broussometia kazinoki (MBK) were investigated. Cell viability of A549 cells was measured by MTT assay. Apoptosis-related protein and MAPK protein levels were measured by Western blot. Chromatin condensation of A549 cells was stained with DAPI. MBK inhibited cell proliferation of A549 cell. Based on DAPI staining, MBK-treated cells manifested nuclear shrinkage, condensation and fragmentation. Treatment of A549 cells with MBK resulted in activation of the caspase-3, -8, -9 and cleavage of poly ADP-ribose polymerase (PARP). In the upstream, MBK increased the expressions Bax and Bak, decreased the expression of Bcl-2, and augmented the Bax/Bcl-2 ratio. MBK-induced apoptosis was accompanied by sustained phosphorylation of JNK, p38 MAPK and apoptosis signal-regulating kinase (ASK)-1. These results suggest that MBK induced apoptosis in A549 cells through Bcl-2 family protein-mediated mitochondria/caspase-3 dependent pathway. In addition, MBK increased the activation of ASK-1, which are critical upsteam signals for JNK/p38 MAPK activation in A549 cancer cells.

• Proceedings of the Korean Nuclear Society Conference
• /
• 2005.10a
• /
• pp.870-871
• /
• 2005

• The Journal of Korean Obstetrics and Gynecology
• /
• v.25 no.3
• /
• pp.16-26
• /
• 2012

Objectives: This study was performed to evaluate effects of Cordyceps militaris(CM) on osteoclast differentiation and its related gene expression. Methods: We used mouse myeloid cells RAW 264.7 stimulated by receptor activator of nuclear factor kappa-B ligand(RANKL) to induce osteoclast differentiation. There are four groups of which RAW 264.7 cells are not stimulated by RANKL (Normal), stimulated by RANKL without CM(Control), stimulated by RANKL with 0.1 ${\mu}g/ml$ of CM(CM 0.1), stimulated by RANKL with 1 ${\mu}g/ml$ of CM(CM 1). Osteoclastogenesis was measured by counting Tartrate-resistant acid phosphatase-positive multinucleated cells [TRAP(+) MNC]. RT-PCR was performed to evaluate the inhibitory effect of CM on gene expression(TRAP, AKT1, JNK1, NFATc1, c-Fos, MITF). Results: 1. CM decreased the number of TRAP(+) osteoclast in RANKL-stimulated RAW 264.7 cell at the concentration of 0.1 ${\mu}g/ml$ and 1 ${\mu}g/ml$. 2. CM decreased the expression of TRAP in osteoclast at the concentration of 1 ${\mu}g/ml$. 3. CM decreased the expression of AKT1, JNK1 in osteoclast at the concentration of 1 ${\mu}g/ml$. 4. CM didn't affect the expression of NFATc1, c-Fos, MITF in osteoclast. Conclusions: Cordyceps militaris has inhibitory effects on osteoclast differentiation and its related gene expression. These results suggest that Cordyceps militaris has a potential as a treatment of osteoporosis.

• Molecules and Cells
• /
• v.27 no.1
• /
• pp.113-118
• /
• 2009

In this study, we have shown the transcriptional regulation of the human GD3 synthase (hST8Sia I) induced by valproic acid (VPA) in human neuroblastoma SK-N-BE(2)-C cells. To elucidate the mechanism underlying the regulation of hST8Sia I gene expression in VPA-stimulated SK-N-BE(2)-C cells, we characterized the promoter region of the hST8Sia I gene. Functional analysis of the 5'-flanking region of the hST8Sia I gene by the transient expression method showed that the -1146 to -646 region, which contains putative binding sites for transcription factors c-Ets-1, CREB, AP-1 and NF-${\kappa}B$, functions as the VPA-inducible promoter of hST8Sia I in SK-N-BE(2)-C cells. Site-directed mutagenesis and electrophoretic mobility shift assay indicated that the NF-${\kappa}B$ binding site at -731 to -722 was crucial for the VPA-induced expression of hST8Sia I in SK-N-BE(2)-C cells. In addition, the transcriptional activity of hST8Sia I induced by VPA in SK-N-BE(2)-C cells was strongly inhibited by SP600125, which is a c-Jun N-terminal kinase (JNK) inhibitor, and $G{\ddot{O}}6976$, which is a protein kinase C (PKC) inhibitor, as determined by RT-PCR (reverse transcription-polymerase chain reaction) and luciferase assays. These results suggest that VPA markedly modulated transcriptional regulation of hST8Sia I gene expression through PKC/JNK signal pathways in SK-N-BE(2)-C cells.