Inhibition of ERK1/2 by silymarin in mouse mesangial cells

The present study aimed to show that pro-inflammatory cytokines [tumor necrosis factor (TNF)-α, interferon (IFN)-γ, and interleukin (IL)-1β] synergistically induce the production of nitric oxide (NO) production in mouse mesangial cells, which play an important role in inflammatory glomerular injury. We also found that co-treatment with cytokines at low doses (TNF-α; 5 ng/ml, IFN-γ; 5 ng/ml, and IL-1β; 1.25 U/ml) synergistically induced NO production, whereas treatment with each cytokine alone did not increase NO production at doses up to 100 ng/ml or 50 U/ml. Silymarin, a polyphenolic flavonoid isolated from milk thistle (Silybum marianum), attenuates cytokine mixture (TNF-α, IFN-γ, and IL-1β)-induced NO production. Western blot and RT-PCR analyses showed that silymarin inhibits inducible nitric oxide synthase (iNOS) expression in a dose-dependent manner. Silymarin also inhibited extracellular signal-regulated protein kinase-1 and -2 (ERK1/2) phosphorylation. Collectively, we have demonstrated that silymarin inhibits NO production in mouse mesangial cells, and may act as a useful anti-inflammatory agent.

Silymarin Inhibits Morphological Changes in LPS-Stimulated Macrophages by Blocking NF-kappaB Pathway

The present study showed that silymarin, a polyphenolic flavonoid isolated from milk thistle (Silybum marianum), inhibited lipopolysaccharide (LPS)-induced morphological changes in the mouse RAW264.7 macrophage cell line. We also showed that silymarin inhibited the nuclear translocation and transactivation activities of nuclear factor-kappa B (NF-kappaB), which is important for macrophage activation-associated changes in cell morphology and gene expression of inflammatory cytokines. BAY-11-7085, an NF-kappaB inhibitor, abrogated LPS-induced morphological changes and NO production, similar to silymarin. Treatment of RAW264.7 cells with silymarin also inhibited LPS-stimulated activation of mitogen-activated protein kinases (MAPKs). Collectively, these experiments demonstrated that silymarin inhibited LPS-induced morphological changes in the RAW264.7 mouse macrophage cell line. Our findings indicated that the most likely mechanism underlying this biological effect involved inhibition of the MAPK pathway and NF-kappaB activity. Inhibition of these activities by silymarin is a potentially useful strategy for the treatment of inflammation because of the critical roles played by MAPK and NF-kappaB in mediating inflammatory responses in macrophages.

Silymarin Inhibits Cytokine-Stimulated Pancreatic Beta Cells by Blocking the ERK1/2 Pathway

We show that silymarin, a polyphenolic flavonoid isolated from milk thistle (Silybum marianum), inhibits cytokine mixture (CM: TNF-alpha, IFN-gamma, and IL-1beta)-induced production of nitric oxide (NO) in the pancreatic beta cell line MIN6N8a. Immunostaining and Western blot analysis showed that silymarin inhibits iNOS gene expression. RT-PCR showed that silymarin inhibits iNOS gene expression in a dose-dependent manner. We also showed that silymarin inhibits extracellular signal-regulated protein kinase-1 and 2 (ERK1/2) phosphorylation. A MEK1 inhibitor abrogated CM-induced nitrite production, similar to silymarin. Treatment of MIN6N8a cells with silymarin also inhibited CM-stimulated activation of NF-kappaB, which is important for iNOS transcription. Collectively, we demonstrate that silymarin inhibits NO production in pancreatic beta cells, and silymarin may represent a useful anti-diabetic agent.

Radicicol Inhibits iNOS Expression in Cytokine-Stimulated Pancreatic Beta Cells

Here, we show that radicicol, a fungal antibiotic, resulted in marked inhibition of inducible nitric oxide synthase (iNOS) transcription by the pancreatic beta cell line MIN6N8a in response to cytokine mixture (CM: TNF-alpha, IFN-gamma, and IL-1beta). Treatment of MIN6N8a cells with radicicol inhibited CM-stimulated activation of NF-kappaB/Rel, which plays a critical role in iNOS transcription, in a dose-related manner. Nitrite production in the presence of PD98059, a specific inhibitor of the extracellular signal-regulated protein kinase-1 and 2 (ERK1/2) pathway, was dramatically diminished, suggesting that the ERK1/2 pathway is involved in CM-induced iNOS expression. In contrast, SB203580, a specific inhibitor of p38, had no effect on nitrite generation. Collectively, this series of experiments indicates that radicicol inhibits iNOS gene expression by blocking ERK1/2 signaling. Due to the critical role that NO release plays in mediating destruction of pancreatic beta cells, the inhibitory effects of radicicol on iNOS expression suggest that radicicol may represent a useful anti-diabetic activity.

Silibinin Inhibits LPS-Induced Macrophage Activation by Blocking p38 MAPK in RAW 264.7 Cells

We demonstrate herein that silibinin, a polyphenolic flavonoid compound isolated from milk thistle (Silybum marianum), inhibits LPS-induced activation of macrophages and production of nitric oxide (NO) in RAW 264.7 cells. Western blot analysis showed silibinin inhibits iNOS gene expression. RT-PCR showed that silibinin inhibits iNOS, TNF-alpha, and IL1beta. We also showed that silibinin strongly inhibits p38 MAPK phosphorylation, whereas the ERK1/2 and JNK pathways are not inhibited. The p38 MAPK inhibitor abrogated the LPS-induced nitrite production, whereas the MEK-1 inhibitor did not affect the nitrite production. A molecular modeling study proposed a binding pose for silibinin targeting the ATP binding site of p38 MAPK (1OUK). Collectively, this series of experiments indicates that silibinin inhibits macrophage activation by blocking p38 MAPK signaling.

Synergistic Induction of iNOS by IFN-gamma and Glycoprotein Isolated from Dioscorea batatas

Dioscorea species continue to be used in traditional Chinese medicine, and represent a major source of steroid precursors for conventional medicine. In the previous study, We isolated glycoprotein (GDB) from Dioscorea batatas, characterized, and demonstrated immunostimulating activity in C57BL/6 mice. The aim of this study was to investigate the mechanism whereby GDB activates macrophages. Macrophages activation by GDB was investigated by analyzing the effects of GDB on nitric oxide (NO) production, iNOS expression, mitogen activated protein kinase (MAPK) phosphorylation, and transcription factor activation. In the presence of IFN-gamma, GDB strongly stimulated macrophages to express iNOS and produce NO. Furthermore, the activation of p38 was synergistically induced by GDB plus IFN-gamma , but SB203580 (a p38 inhibitor) inhibited GDB plus IFN-gamma-induced p38 activation. This study indicates that GDB is an important activator of macrophages. Furthermore, due to the critical role that macrophage activation plays in innate immune response, the activation effects of GDB on macrophages suggest that GDB may be a useful immunopotentiating agent.

Characterization and Immunopotentiating Effects of the Glycoprotein Isolated from Dioscorea Batatas

We demonstrate that glycoprotein isolated from Dioscorea batatas (GDB) has immunostimulatory effects including macrophage activation. Analysis of infiltration of inflammatory cells into peritoneal cavity showed GDB treatment significantly increased the recruitment of macrophages, lymphocytes, neutrophils, and monocytes into the peritoneal cavity. Treatment of spleen cells isolated from C57BL/6 mice with GDB significantly increased the proliferation of B cells and T cells induced by LPS and ConA, respectively. Treatment with GDB significantly increased the cytolytic capacity of NK cells and macrophages against YAC-1 and B16 cells, respectively. In order to further confirm and investigate the mechanism of GDB on macrophage activation, we analyzed the effects of GDB on the cytokine expression including iNOS, IL-1beta, and TNF-alpha in mouse macrophage cell line, RAW 264.7 cells. RT-PCR and ELISA showed that GDB increased the expression of IL-1beta, and TNF-alpha, whereas iNOS was not induced by GDB. Collectively, this series of experiments indicates that GDB stimulates immune system including macrophage activation.

Magnolol Inhibits LPS-induced NF-kappaB/Rel Activation by Blocking p38 Kinase in Murine Macrophages

This study demonstrates the ability of magnolol, a hydroxylated biphenyl compound isolated from Magnolia officinalis, to inhibit LPS-induced expression of iNOS gene and activation of NF-kappaB/Rel in RAW 264.7 cells. Immunohisto-chemical staining of iNOS and Western blot analysis showed magnolol to inhibit iNOS gene expression. Reporter gene assay and electrophoretic mobility shift assay showed that magnolol inhibited NF-kappaB/Rel transcriptional activation and DNA binding, respectively. Since p38 is important in the regulation of iNOS gene expression, we investigated the possibility that magnolol to target p38 for its anti-inflammatory effects. A molecular modeling study proposed a binding position for magnolol that targets the ATP binding site of p38 kinase (3GC7). Direct interaction of magnolol and p38 was further confirmed by pull down assay using magnolol conjugated to Sepharose 4B beads. The specific p38 inhibitor SB203580 abrogated the LPS-induced NF-kappaB/Rel activation, whereas the selective MEK-1 inhibitor PD98059 did not affect the NF-kappaB/Rel. Collectively, the results of the series of experiments indicate that magnolol inhibits iNOS gene expression by blocking NF-kappaB/Rel and p38 kinase signaling.

Inhibition of p65 Nuclear Translocation by KIOM-79 / 대한해부학회지

We demonstrate that KIOM-79, combined extracts isolated from Magnolia officinalis, Pueraria lobata, Glycyrrhiza uralensis, and Euphorbia pekinensis, inhibits LPS-induced expression of iNOS gene in RAW 264.7 cells. Treatment of RAW 264.7 cells with KIOM-79 inhibited LPS-stimulated nitric oxide production in a doserelated manner. Immunohisto-chemical staining of iNOS and RT-PCR analysis showed that the decrease of NO was due to the inhibition of iNOS gene expression. Immunostaining of p65 and EMSA showed that KIOM-79 inhibited NF-kappa/Rel nuclear translocation and DNA binding, respectively. Collectively, this series of experiments indicates that KIOM inhibits iNOS gene expression by blocking NF-kappa/Rel. Due to the critical role that NO release plays in mediating inflammatory responses, the inhibitory effects of KIOM-79 on iNOS suggest that KIOM-79 may represent a useful anti-inflammatory agent.

iNOS Induction by Polysaccharide Isolated from Astragalus membranaceus / 체질인류학회지

Astragalus membranaceus is used as a natural herbal medicine in East Asia for preventing carcinogenesis and reducing side effects induced by chemotherapy in cancer patients. Although the mechanism of anti-tumor activity is not known, the polysaccharides may potentiate the host defense mechanism through the activation of immune system. The objective of this study is to investigate the mechanism by which APS activates macrophages. To analyze macrophage activation and iNOS gene expression, we performed nitrite generation assay, immunohistochemistry, and RT-PCR. In the present study we show that a polysaccharide isolated from the Astragalus membranaceus (Astragalus Polysaccharide, APS) significantly induces nitric oxide (NO). Immunohistochemical staining of inducible NO synthase (iNOS) showed that the increase of NO was due to the induction of iNOS production. To further study the mechanism responsible for the induction of iNOS, we investigated the effect of APS on the iNOS mRNA expression. RT-PCR analysis showed that APS produced significant induction of iNOS gene expression. In conclusion, we demonstrate that a polysaccharide isolated from Astragalus membranaceus stimulates macrophages to generate NO through the activation of iNOS gene expression.

Nuclear Translocation of NF-kappaB/Rel Component by PCSC22 Isolated from Poria cocos Sclerotium / 대한해부학회지

The sclerotium of Poria cocos Wolf, which grows on the roots of pine trees, has long been used as a sedative and diuretic (Chang and But, 1987). The accumulating data revealed that certain ingredients of the sclerotium of Poria cocos showed anti-tumor activities (Kanayama, 1986). Although the mechanism of anti-tumor activity is not known, the polysaccharides may potentiate the host defense mechanism through the activation of immune system. In the present study we show that PCSC22, a polysaccharide isolated from the sclerotium of Poria cocos with one percent sodium carbonate, significantly induces nitric oxide (NO) production and inducible NO synthase (iNOS) transcription. To further investigate the mechanism responsible for the induction of iNOS gene expression, we investigated the effect of PCSC22 on the activation of NF-kappaB/Rel, whose binding site was located in the promoter of iNOS gene. Immuno-histo-chemical staining of p65 and p50 showed that PCSC22 produced strong induction of NF-kappaB/Rel nuclear translocation. Electrophoretic mobility shift assay (EMSA) further confirmed the activation of NF-kappaB/Rel by PCSC22. In conclusion, we demonstrate that PCSC22 stimulates macrophages to express iNOS gene through the activation of NF-kappa B/Rel.

Activation of Macrophage by PCSC22 Isolated from Poria cocos Sclerotium / 체질인류학회지

The sclerotium of Poria cocos Wolf, which grows on the roots of pine trees, has long been used as a sedative, diuretic, and anti-inflammatory agent. The accumulating data revealed that certain ingredients of the sclerotium of Poria cocos showed anti-tumor activities. Although the mechanism of anti-tumor activity is not known, the polysaccharides may potentiate the host defense mechanism through the activation of immune system. In the present study we show that PCSC22, a polysaccharide isolated from the sclerotium of Poria cocos with one percent sodium carbonate, significantly induces nitric oxide (NO). Immunohistochemical staining of inducible NO synthase (iNOS) showed that the increase of NO was due to the induction of iNOS production. To further study the mechanism responsible for the induction of iNOS gene expression, we investigated the effect of PCSC22 on the activation of p38 kinase, which is important in the gene expression of inflammatory cytokines including iNOS. Western blot assay showed that PCSC22 produced phosphorylation of p38 kinase. In conclusion, we demonstrate that PCSC stimulates macrophages to express iNOS gene through the activation of p38 kinase.

The correlation between dental compensation and craniofacial morphology in skeletal Class III malocclusion / 대한치과교정학회지

This investigation was designed to analyze the degree of dental compensation according to horizontal components of craniofacial skeleton and to investigate correlation between dental compensation and craniofacial pattern in skeletal class III malocclusion. The material selected for this study consisted of standard lateral cephalogram of 59 subjects in normal occlusion group, 91 subjects in mild skeletal class III malocclusion group and 58 subjects in severe skeletal class III malocclusion group. The mild skeletal class III malocclusion group was divided into two groups, one was class III malocclusion without anterior crossbite group and the other was class III malocclusion with anterior crossbite group. The data were analyzed by Quick-ceph image program. The results were as follows. 1. Mild skeletal class III malocclusion without anterior crossbite group showed the most labial inclination of upper incisors, followed by severe skeletal class III malocclusion group and mild skeletal class III malocclusion with anterior crossbite group, the Latter showing the least. The amount of lingual inclination of lower incisors was the largest in severe skeletal class III malocclusion group, and there was no statistically significant difference between mild skeletal claw III malocclusion without anterior crossbite group and mild skeletal class III malocclusion with anterior crossbite group. 2. There were little differences in vertical skeletal structure between mild skeletal class III malocclusion without anterior crossbite group and mild skeletal class III malocclusion with anterior cwssbite group, they showed statistically significant differences in the upper incisors measurements. 3. The measurements of lower incisors in mild skeletal class III malocclusion without anterior crossbite group and upper incisors in mild skeletal class III malocclusion with anterior crossbite group represented a high correlation with skeletal structure. Especially, deltaIMPA and deltaFMIA of lower incisor measurements, and deltaU1-FR deltaUi-SN of upper incisor measurements showed high correlation with skeletal structure in each group. 4. deltaIMPA and deltaFMIA of lower incisor measurements showed high correlation with skeletal structure in all groups. deltaUI-FH, deltaU1-SN and Ui-facial plane(mm) of upper incisor measurements represented higher correlation with skeletal structure than any other upper incisor measurements.

Current status of general thoracic surgery in Korea / 대한흉부외과학회지

No abstract available.