Hyo-Jin An

Reduced IL-2 but elevated IL-4, IL-6, and IgE serum levels in patients with cerebral infarction during the acute stage

Cytokines in the central nervous system (CNS) may play an important role in functioning as intercellular signals that orchestrate the response to injury. Whether this is a cause or result of the brain disease process is uncertain. We investigated IFN-γ, IL-2, IL-4, IL-6, and IgE in the sera of 38 patients with cerebral infarction during the acute stage and 10 normal controls using an originally devised sensitive sandwich enzyme-linked immunosorbent assay (ELISA). We found that serum levels of IL-2 derived from T helper 1 (Th1) cells were slightly reduced in patients with cerebral infarction, whereas serum levels of IL-4 and IL-6 derived from Th2 cells were elevated significantly. IL-4 induces synthesis of IgE in human B cells. Endogenous IL-6 plays an obligatory role in IL-4-dependent human IgE synthesis. We observed that serum IgE levels were elevated significantly in patients with cerebral infarction. However, serum IFN-γ levels were not elevated significantly in cerebral infarction patients. These findings suggest that elevated IL-4, IL-6, and IgE levels in the human serum may be an important factor in cerebral infarction during the acute stage. Decrease of IL-2 levels in the serum of patients with cerebral infarction may be a regulatory mechanism.

Tormentic acid, a triterpenoid saponin, isolated from Rosa rugosa, inhibited LPS-induced iNOS, COX-2, and TNF-α expression through inactivation of the nuclear factor-κb pathway in RAW 264.7 macrophages

We previously reported that extract of Rosa rugosa root and its active triterpenoids constituents exhibit anti-nociceptive and anti-inflammatory effects in animal models. However, little is known about the effects and the molecular mechanism of the 19α-hydroxyursane-type triterpenoids. Among the tested 19α-hydroxyursane-type triterpenoids (kaji-ichigoside F(1), rosamultin, euscaphic acid, tormentic acid (TA)), TA was found to most potently inhibit the production of nitric oxide (NO) in RAW 264.7 cells. We investigated the anti-inflammatory effects and its underlying molecular mechanisms of TA in lipopolysaccaride (LPS)-stimulated RAW 264.7 cells. TA dose-dependently reduced the productions of NO, prostaglandin E(2) (PGE(2)), and tumor necrosis factor-α (TNF-α) induced by LPS. In addition, TA significantly suppressed the LPS-induced expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and TNF-α at the mRNA and protein levels. Moreover, treatment with TA decreased LPS-induced DNA binding of nuclear factor-kappa B (NF-κB) and nuclear translocation of p65 and p50 subunits of NF-κB. Consistent with these findings, TA also suppressed the LPS-stimulated degradation and phosphorylation of inhibitor of kappa B-α (IκB-α). Taken together, these results suggest that the anti-inflammatory activity of TA is associated with the down-regulation of iNOS, COX-2, and TNF-α through the negative regulation of the NF-κB pathway in RAW 264.7 cells.

Inhibitory effects of Rumex japonicus Houtt. on the development of atopic dermatitis-like skin lesions in NC/Nga mice.

Background  Rumex japonicus Houtt. (RJH) is one of the herbs used in Eastern countries for the treatment of atopic dermatitis (AD). It has been shown to have an antioxidative effect in human skin disease.

Use of Electroacupuncture at ST36 to Inhibit Anaphylactic and Inflammatory Reaction in Mice

Objective: Electroacupuncture (EA) has been used to treat myalgia, allergy and gastroenteropathy in Korea. To determine whether EA can treat anaphylactic and inflammatory reactions, the effect of EA was investigated in a murine model. Methods: EA stimulation of the ST36 acupoint was performed for 10 min. Using a passive cutaneous anaphylaxis (PCA) model, the antianaphylactic effects of EA were examined. Interleukin-6 and tumor necrosis factor-α were measured using the ELISA method. The level of nuclear factor (NF)-ĸB/RelA protein and NF-ĸB DNA-binding activity was determined using the Western blot analysis and the transcription factor enzyme-linked immunoassay method. Results: EA inhibits PCA and β-hexosaminidase release, IL-6 secretion on the PCA, and in addition, EA reduces NF-ĸB DNA-binding activity. Conclusion: These results indicate that EA may possess antianaphylactic and antiinflammatory properties.

Roxatidine suppresses inflammatory responses via inhibition of NF‐κB and p38 MAPK activation in LPS‐induced RAW 264.7 macrophages

Roxatidine is a novel, specific, competitive H2‐receptor antagonist that is used to treat gastric and duodenal ulcers, and which is known to suppress the growth of several tumors by reducing vascular endothelial growth factor (VEGF) expression. Nevertheless, it remains unclear whether roxatidine has anti‐inflammatory effects. In this study, we the authors investigated the anti‐inflammatory effect of roxatidine in lipopolysaccharide (LPS)‐stimulated RAW 264.7 macrophage cells. It was found that roxatidine dose‐dependently inhibited the productions of prostaglandin E2 (PGE2), nitric oxide (NO), and histamine, and the protein and mRNA expressions of cyclooxygenase‐2 (COX‐2), inducible nitric oxide synthase (iNOS), and histidine decarboxylase (HDC). In addition, roxatidine reduced the productions and expressions of VEGF‐1 and pro‐inflammatory cytokines, including those of tumor necrosis factor‐α (TNF‐α), interleukin‐1β (IL‐1β), and interleukin‐6 (IL‐6). Electrophoretic mobility shift assays (EMSA) and reporter gene assays revealed that treatment with roxatidine attenuated the LPS‐induced DNA‐binding and transcriptional activity of nuclear factor kappa B (NF‐κB). In addition, it was found that pretreatment with roxatidine significantly inhibited the nuclear translocations of the p65 and p50 subunits of NF‐κB, and these inhibitions were not found to be associated with decreases in the phosphorylation or degradation of inhibitory kappa B‐α (IκBα). Furthermore, roxatidine suppressed the phosphorylation of p38 MAP kinase, but not of IκB kinase‐α/β (IKKα/β), c‐Jun NH2‐terminal kinase (JNK), or extracellular signal‐regulated kinase (ERK). Taken together, these results indicate that the anti‐inflammatory properties of roxatidine in LPS‐treated RAW 264.7 macrophages are mediated by the inhibition of NF‐κB transcriptional activity and the p38 MAP kinase pathway. J. Cell. Biochem. 112: 3648–3659, 2011.

Water extract isolated from Chelidonium majus enhances nitric oxide and tumour necrosis factor-α production via nuclear factor-κB activation in mouse peritoneal macrophages

Chelidonium majus is used to treat several inflammatory diseases and tumours. We have examined the effect of C. majus on nitric oxide (NO) production using mouse peritoneal macrophages. When C. majus was used in combination with recombinant interferon‐γ (rIFN‐γ, 10U mL−1), there was a marked cooperative induction of NO production. Treatment of rIFN‐γ plus C. majus (1 mg mL−1) in macrophages caused a significant increase in tumour necrosis factor‐α (TNF‐α) production. The increased production of NO and TNF‐α from rIFN‐γ plus C. majus‐stimulated cells was almost completely inhibited by nuclear factor‐κB (NF‐κB) inhibitor, pyrrolidine dithiocarbamate (100 μM). These findings demonstrated that C. majus increased the production of NO and TNF‐α by rIFN‐γ‐primed macrophages and suggested that NF‐κB played a critical role in mediating the effects of C. majus.

Immunostimulatory activity of polysaccharides from Cheonggukjang

Cheonggukjang is a Korean whole soybean paste fermented by Bacillus subtilis and regarded as a healthy food. The objective of this study was to investigate the immunostimulatory activity of polysaccharides from Cheonggukjang (PSCJ) in RAW 264.7 macrophages and an animal model. PSCJ induced mRNA expressions of inducible nitric oxide synthase and tumor necrosis factor-α (TNF-α) by activating nuclear factor-κB, and subsequently increased the productions of nitric oxide (NO) and TNF-α in murine recombinant interferon-γ-primed RAW 264.7 macrophages. Furthermore, after daily oral administration of PSCJ, immobility time decreased significantly in the PSCJ-administered group (200 or 400 mg/kg) on day 10. Taken together, these results suggest that the PSCJ has a possible role improving immune function through regulatory effects on immunological parameters, such as NO and TNF-α productions and changes in indicators related to fatigue.

Mountain Grown Ginseng Induces Apoptosis in HL-60 Cells and Its Mechanism Have Little Relation with TNF-α Production

The root of ginseng is one of the most popular natural tonics in Oriental countries. Ginseng grown in the wild, deep in the mountains, is known as Sansam (mountain grown ginseng, MGG). MGG belongs to Araliaceae and Panax. In this study, we investigated the effects of MGG on the cytotoxicity, induction of apoptosis and the putative pathways of its actions in human promyelocytic leukemia cells, HL-60. Using apoptosis analysis, we found that MGG is a potent inducer of apoptosis, but it has less effect on human peripheral blood mononuclear cells. Caspase-3 activation and subsequent apoptotic cell death in MGG-treated cells were partially blocked by the caspase-3 inhibitor, Z-DEVD-FMK. MGG also inhibited the caspase-8 activity. To determine whether MGG-induced apoptosis is involved in tumor necrosis factor-alpha (TNF-alpha) secretion, TNF-alpha secretion was quantified by enzyme-linked immunosorbent assay (ELISA) method. Unexpectedly, MGG significantly decreased the TNF-alpha secretion compared to the control. These results suggest that MGG-induced cytotoxicity have little relation with the secretion of TNF-alpha in HL-60 cells. Furthermore, MGG with rIFN-gamma synergistically increased nitric oxide (NO) production in mouse peritoneal macrophages. Taken together, our data indicate that MGG is a potent inducer of apoptosis on HL-60 cells and these abilities could be used clinically for the treatment of cancer.

The bark of Betula platyphylla var. japonica inhibits the development of atopic dermatitis-like skin lesions in NC/Nga mice

The bark of Betula platyphylla Sukat. var. japonica Hara (Asian White Birch, AWB) is one of the herbs used in Eastern countries for the treatment of various inflammatory diseases including atopic dermatitis (AD). The present study was performed to examine if and how the bark of AWB inhibits the development of AD-like skin lesions in NC/Nga mice induced by repeated application of picryl chloride (PC). With this aim, we examined the skin symptom severity, itching behavior, serum immunoglobulin (Ig) E level and mRNA expression of cytokines at iliac and cervical lymph nodes in the mice. Oral administration of AWB extracts (25, 100 and 250 mg/kg) to the PC-treated mice inhibited the development of AD-like skin lesions as exemplified by a significant decrease in the total skin severity scores, itching behavior and a decrease in hypertrophy and infiltration of inflammatory cells into dermis. The serum IgE level was also significantly reduced by AWB extract. In the RT-PCR results, the expression of interleukin-4 mRNA was reduced by AWB extract, whereas the expression of interferon-gamma mRNA was not changed. These results suggest that AWB inhibits the development of AD-like skin lesions in NC/Nga mice through the suppression of the T-helper 2 cell response.

Molecular mechanisms underlying the anti-obesity potential of prunetin, an O-methylated isoflavone

Prunetin is an O-methylated isoflavone, which is a type of flavonoid. There are a limited number of reports detailing the biological activities of prunetin. Although an anti-inflammatory effect of prunetin has been reported in vitro, to our knowledge, there have been no reports on anti-adipogenic effects of prunetin in obese animals. The aims of this study were to determine whether prunetin suppresses high-fat diet (HFD)-induced adipogenesis in the liver and visceral adipose tissues of mice, and to explore the underlying mechanisms mediating the actions of prunetin. To this end, mice were fed a HFD for 10 weeks to induce obesity, and prunetin (10 μg/kg or 20 μg/kg) was administered in the last 3 weeks. Compared to saline-treated mice, mice treated with prunetin showed significantly reduced body weight gain, visceral fat pad weights, and plasma glucose levels. We found that prunetin significantly inhibited the HFD-induced upregulation of the expression of important adipogenic genes (PPARγ, C/EBPα, SREBP, aP2, LPL adiponectin, and leptin), and suppressed HFD-mediated increase in expression of lipid metabolism-related genes (SREBP, PPARγ, LXR, and HMG-CoA) in the liver tissues. Furthermore, prunetin induced expression of adiponectin receptors 1 and 2 (adipoR1, adipoR2), as well as that of AMP-activated protein kinase (AMPK) in the liver and adipose tissue. These results suggest that prunetin mediates anti-obesity/adipogenesis effects by suppressing obesity-related transcription through a feedback mechanism that regulates the expression of adiponectin, adipoR1, adipoR2, and AMPK.