• The Korea Journal of Herbology
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• v.23 no.3
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• pp.119-125
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• 2008

Objectives : The purpose of this paper was to investigate the anti-inflammatory effects of extract from Teucrium veronicoides (TV) on the RAW 264.7 cells. Methods : To evaluate of anti-inflammatory of TV, we examined the cytokine productions on lipopolysacchride (LPS)-induced RAW 264.7 cells and also inhibitory mechanisms using Western blot. Furthermore, We examined LPS-induced endotoxin shock. Results : 1. Extract from TV itself does not have any cytotoxic effect. 2. Extract from TV reduced LPS-induced Nitric oxide (NO),interleukin (IL)-1b, IL-6 and IL-10, tumor necrosis factor-a (TNF-a) production in RAW 264.7 cells. 3. TV inhibited the activation of mitogen-activated protein kinases (MAPKs) such as p38, extracelluar signal-regulated kinase (ERK 1/2) and c-Jun NH2-terminal kinase (JNK) and also the degradation of inhibitory kappa B a (Ik-Ba) in the LPS-stimulated RAW 264.7 cells. 3. TV slightly increased the duration of survival after LPS-induced endotoxin shock. Conclusions : TV down-regulated LPS-induced NO and cytokines production, which could provide a clinical basis for anti-inflammatory properties of TV.

• Journal of Life Science
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• v.24 no.9
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• pp.927-934
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• 2014

Sanguinarine, a benzophenanthridine alkaloid originally derived from the root of Sanguinaria canadensis, has been shown to possess antimicrobial, antioxidant, and anti-cancer properties. Although tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is known to induce apoptosis in cancer cells, but not most normal cells and has shown efficacy in a phase 2 clinical trial, development of resistance to TRAIL by tumor cells is a major roadblock. Our previous study indicated that treatment with TRAIL in combination with subtoxic concentrations of sanguinarine sensitized TRAIL-mediated apoptosis in TRAIL-resistant human gastric carcinoma AGS cells; however, the detailed mechanisms are not fully understood. In this study, we show that sanguinarine sensitizes AGS cells to TRAIL-mediated apoptosis as detected by MTT assay, agarose gel electrophoresis, chromatin condensation and flow cytometry analysis. Combined treatment with sanguinarine and TRAIL effectively induced expression of death receptor (DR) 5 but did not affect expression of DR4 and mitogen activated protein kinases signaling molecules. Moreover, the combined treatment with sanguinarine and TRAIL increased the generation of reactive oxygen species (ROS); however, N-acetylcysteine, ROS scavenger, significantly recovered growth inhibition induced by the combined treatment. Taken together, our results indicate that sanguinarine can potentiate TRAIL-mediated apoptosis through upregulation of DR5 expression and ROS generation.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.11
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• pp.1300-1307
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• 2017

Cheonggukjang is well known as a traditional fermented food in Korea and has various biological activity. In this study, immune-enhancing activity of extract of cheonggukjang fermented with Bacillus amyloliquefaciens (SRCM100730) was examined in RAW 264.7 murine macrophages. Treatment with extract augmented production of nitric oxide (NO) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) from RAW 264.7 macrophages in a dose-dependent manner. Similarly, increased mRNA expression of inducible nitric oxide synthase (iNOS) and $TNF-{\alpha}$ was observed. In addition, the extract synergistically enhanced production of NO and $TNF-{\alpha}$ from lipopolysaccharide (LPS)-stimulated macrophages. Analysis of intracellular pathways revealed that the immune-enhancing activity of cheonggukjang extract was related to activation of mitogen-activated protein kinases (MAPK) and nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$). These results suggest that cheonggukjang fermented with B. amyloliquefaciens (SRCM100730) is a beneficial food effective for activation of immune responses.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.6
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• pp.1392-1398
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• 2009

The purpose of this study was to investigate the anti-inflammatory effects of aqueous extract from Sophora Japonica (SJ) on the RAW 264.7 cells. To evaluate the anti-inflammatory effects of SJ, we examined the cytokine productions including nitric oxide (NO), interleukin (IL)-1b, IL-6 and tumor necrosis factor-a (TNF-a) in lipopolysaccharide (LPS)-induced RAW 264.7 cells and also inhibitory mechanisms such as mitogen-activated protein kinases (MAPKs) and nuclear factor kappa b (NF-kB) using Western blot. SJ inhibited LPS-induced production of NO, TNF-a but not of IL-1b and IL-6 in RAW 264.7 cells. SJ inhibited the activation of MAPKs such as extracelluar signal-regulated kinase (ERK 1/2), c-Jun NH2-terminal kinase (JNK) and p38 but not of NF-kB in the LPS-stimulated RAW 264.7 cells. In conclusion, SJ down-regulated LPS-induced NO and TNF-a productions via MAPKs, which could be a clinical basis for inflammatory diseases and autoimmune diseases.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.5
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• pp.1211-1216
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• 2006

The effect of a chronic programme of either low- or moderate-to-high-intensity treadmill running on the activation of the Extracellular-signal regulated protein kinase (ERK1/2), Phosphorylated ERK 1/2(pERK1/2) and the Phosphorylated c-Jun N-terminal kinase(pJNK) pathways was determined in rat Back skin Hair follicle. Sprague-Dawley rats were assigned to one of three groups: (i) sedentary group(NE; n=10); (ii) low-intensity exercise group (Bm/min; LIE; n=10); and (iii) moderate-high-intensity exercise group(28m1min; HIE; n=10). The training regimens were planned so that animals covered the same distance and had similar utilization for both LIE and HIE exercise sessions. The report runs as follows; A single bout of LIE or HIE following 4 weeks of exercise led to a twofold increase in the phosphorylation of ERK2, pERK2 and a threefold increase in pJNKl, pERKl. ERKI phosphorylation in LIE Back skin sampled and pJNK2 in HIE Back skin sampled 48h after the last exercise bout was similar to sedentary values, while pJNK2 phosphorylation in LIE Back skin sampled was 70-80% lower than sedentary. 48h after the last exercise bout of LIE or HIE increased ERK2, pERKl and pJNKl expression, with the magnitude of this increase being independent of prior exercise intensity or duration. PERK1/2, pJNKl expression was increased Three- to fourfold in Back skin Hair follicle sampled 48h after the last exercise bout irrespective of the prior exercise programme, but ERKI expression in HIE Back skin sampled was approximately 90% lower than sedentary values. In conclusion, exercise-training of different jntensities/durations results in selective postexercise activation of intracellular signal pathways, which may be one mechanism regulating specific adaptations induced by diverse training programmes.

• Molecules and Cells
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• v.21 no.1
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• pp.42-51
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• 2006

We investigated the mechanism of contraction induced by S1P in esophageal smooth muscle cells. Western blot analysis demonstrated that $S1P_1$, $S1P_2$, $S1P_3$, and $S1P_5$ receptors existed in the cat esophagus. Only penetration of EDG-5 ($S1P_2$) antibody into permeabilized cells inhibited S1P-induced contraction. Pertussis toxin (PTX) also inhibited contraction, suggesting that it was mediated by $S1P_2$ receptors coupled to a PTXsensitive $G_i$ protein. Specific antibodies to $G_{i2}$, $G_q$ and $G_{\beta}$ inhibited contraction, implying that the S1P-induced contraction depends on PTX-insensitive $G_q$ and $G_{\beta}$ dimers as well as the PTX-sensitive $G_{i2}$. Contraction was not affected by the phospholipase $A_2$ inhibitor DEDA, or the PLD inhibitor ${\rho}$-chloromercuribenzoate, but it was abolished by the PLC inhibitor U73122. Incubation of permeabilized cells with $PLC{\beta}3$ antibody also inhibited contraction. Contraction involved the activation of a PKC pathway since it was affected by GF109203X and chelerythrine. Since $PKC{\varepsilon}$ antibody inhibited contraction, $PKC{\varepsilon}$ may be required. Preincubation of the muscle cells with the MEK inhibitor PD98059 blocked S1P-induced contraction, but the p38 MAP kinase inhibitor SB202190 did not. In addition, co-treatment of cells with GF 109203X and PD98059 did not have a synergistic effect, suggesting that these two kinases are involved in the same signaling pathway. Our data suggest that S1P-induced contraction in esophageal smooth muscle cells is mediated by $S1P_2$ receptors coupled to PTX-sensitive $G_{i2}$ proteins, and PTX-insensitive $G_q$ and $G_{\beta}$ proteins, and that the resulting activation of the $PLC{\beta}3$ and $PKC{\varepsilon}$ pathway leads to activation of a p44/p42 MAPK pathway.

• Journal of Life Science
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• v.31 no.2
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• pp.115-125
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• 2021

Calcium-dependent protein kinases (CDPKs) are known to be involved in regulating plant responses to abiotic stresses such as salinity, cold temperature and dehydration,. Although CDPKs constitute a large multigene family consisting of 31 genes in rice, only a few rice CDPKs' functions have been identified. Therefore, in order to elucidate the functions of OsCPK11 in rice, this study was intended to focus on the expression pattern analysis of OsCPK11 in wild type and ND0001 oscpk11 mutant plants under these abiotic stresses. For the salt, cold and drought stress treatment, seedlings were exposed to 200 mM NaCl, 4℃ and 20% PEG 6,000, respectively. RT-PCR and quantitative real-time PCR were performed to determine the expression patterns of OsCPK11 in wild type and ND0001 mutant plants. RT-PCR results showed that OsCPK11 transcripts in the wild type and heterozygous mutant were detected, but not in the homozygous mutant. Real-time PCR results showed that relative expression of OsCPK11 of wild type plants was increased and reached to the highest level at 24 hr, at 6 hr and at 24 hr under salt, cold and drought stress conditions, respectively. Relative expression of OsCPK11 of ND0001 homozygous plant was significantly reduced compared to that of wild type. These results suggested that oscpk11 homozygous mutant knocks out OsCPK11 and OsCPK11 might be involved in salt, cold and drought stress signaling by regulating its gene expression.

• Journal of Life Science
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• v.31 no.5
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• pp.464-474
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• 2021

Inflammation induced by metabolic syndromes, cancers, injuries, and sepsis can alter cellular metabolism by reducing mitochondrial function via oxidative stress, thereby resulting in neuropathy and muscle atrophy. In this study, we investigated whether butyrate, a short chain fatty acid produced by gut microbiota, could prevent mitochondrial dysfunction and muscle atrophy induced by lipopolysaccharide (LPS) in the C2C12 cell line. LPS-activated MAPK signaling pathways increased the levels of the mitochondrial fission signal, p-DRP1 (Ser616), and the muscle atrophy marker, atrogin 1. Interestingly, butyrate significantly inhibited the phosphorylation of JNK and p38 and reduced the atrogin 1 level in LPS-treated C2C12 cells while increasing the phosphorylation of DRP1 (Ser637) and levels of mitofusin2, which are both mitochondrial fusion markers. Next, we investigated the effect of MAPK inhibitors, finding that butyrate had the same effect as JNK inhibition in C2C12 cells. Also, butyrate inhibited the LPS-induced expression of pyruvate dehydrogenase kinase 4 (PDK4), resulting in decreased PDHE1α phosphorylation and lactate production, suggesting that butyrate shifted glucose metabolism from aerobic glycolysis to oxidative phosphorylation. Finally, we found that these effects of butyrate on LPS-induced mitochondrial dysfunction were caused by its antioxidant effects. Thus, our findings demonstrate that butyrate prevents LPS-induced muscle atrophy by improving mitochondrial dynamics and metabolic stress via the inhibition of JNK phosphorylation. Consequently, butyrate could be used to improve LPS-induced mitochondrial dysfunction and myopathy in sepsis.

• Biomolecules & Therapeutics
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• v.29 no.2
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• pp.211-219
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• 2021

Alopecia is a distressing condition caused by the dysregulation of anagen, catagen, and telogen in the hair cycle. Dermal papilla cells (DPCs) regulate the hair cycle and play important roles in hair growth and regeneration. Myristoleic acid (MA) increases Wnt reporter activity in DPCs. However, the action mechanisms of MA on the stimulation of anagen signaling in DPCs is not known. In this study, we evaluated the effects of MA on anagen-activating signaling pathways in DPCs. MA significantly increased DPC proliferation and stimulated the G2/M phase, accompanied by increasing cyclin A, Cdc2, and cyclin B1. To elucidate the mechanism by which MA promotes DPC proliferation, we evaluated the effect of MA on autophagy and intracellular pathways. MA induced autophagosome formation by decreasing the levels of the phospho-mammalian target of rapamycin (phospho-mTOR) and increasing autophagy-related 7 (Atg7) and microtubule-associated protein 1A/1B-light chain 3II (LC3II). MA also increased the phosphorylation levels of Wnt/β-catenin proteins, such as GSK3β (Ser9) and β-catenin (Ser552 and Ser675). Treatment with XAV939, an inhibitor of the Wnt/β-catenin pathway, attenuated the MA-induced increase in β-catenin nuclear translocation. Moreover, XAV939 reduced MA-induced effects on cell cycle progression, autophagy, and DPC proliferation. On the other hand, MA increased the levels of phospho (Thr202/Tyr204)-extracellular signal regulated kinases (ERK). MA-induced ERK phosphorylation led to changes in the expression levels of Cdc2, Atg7 and LC3II, as well as DPC proliferation. Our results suggest that MA promotes anagen signaling via autophagy and cell cycle progression by activating the Wnt/β-catenin and ERK pathways in DPCs.

• Animal Bioscience
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• v.35 no.7
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• pp.989-998
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• 2022

Objective: This study investigated the effects of intrauterine growth restriction (IUGR) during late pregnancy on the cell growth, proliferation, and differentiation in ovine fetal thymuses. Methods: Eighteen time-mated Mongolian ewes with singleton fetuses were allocated to three groups at d 90 of pregnancy: restricted group 1 (RG1, 0.18 MJ ME/body weight [BW]^0.75/d, n = 6), restricted group 2 (RG2, 0.33 MJ ME/BW^0.75/d, n = 6) and control group (CG, ad libitum, 0.67 MJ ME/BW^0.75/d, n = 6). Fetuses were recovered at slaughter on d 140. Results: The G0/G1 phase cell number in fetal thymus of the RG1 group was increased but the proliferation index and the expression of proliferating cell nuclear antigen (PCNA) were reduced compared with the CG group (p<0.05). Fetuses in the RG1 group exhibited decreased growth hormone receptor (GHR), insulin-like growth factor 2 receptor (IGF-2R), and their mRNA expressions (p<0.05). For the RG2 fetuses, there were no differences in the proliferation index and PCNA expression (p>0.05), but growth hormone (GH) and the mRNA expression of GHR were lower than those of the CG group (p<0.05). The thymic mRNA expressions of cyclin-dependent protein kinases (CDKs including CDK1, CDK2, and CDK4), CCNE, E2-factors (E2F1, E2F2, and E2F5) were reduced in the RG1 and RG2 groups (p<0.05), and decreased mRNA expressions of E2F4, CCNA, CCNB, and CCND were occurred in the RG1 fetuses (p<0.05). The decreased E-cadherin (E-cad) as a marker for epithelial-mesenchymal transition (EMT) was found in the RG1 and RG2 groups (p<0.05), but the OB-cadherin which is a marker for activated fibroblasts was increased in fetal thymus of the RG1 group (p<0.05). Conclusion: These results indicate that weakened GH/IGF signaling system repressed the cell cycle progression in G0/G1 phase in IUGR fetal thymus, but the switch from reduced E-cad to increased OB-cadherin suggests that transdifferentiation process of EMT associated with fibrogenesis was strengthened. The impaired cell growth, retarded proliferation and modified differentiation were responsible for impaired maturation of IUGR fetal thymus.