Protective Effects of Spatholobi Caulis Extract on Neuronal Damage and Focal Ischemic Stroke/Reperfusion Injury.

Neuronal apoptotic cell death plays an important role in many neurological disorders, including Alzheimer's disease, Parkinson's disease, and ischemic stroke. Spatholobi Caulis (SC) has been widely used in traditional herbal medicine for the treatment of cancer, inflammation, viral infection, and anemia. However, the protective effects of SC extract (SCE) against apoptotic cell death in the brain have not been reported. We investigated the protective effects of SCE against neuronal injury etoposide-induced neurotoxicity and in rats subjected to focal transient ischemic stroke middle cerebral artery occlusion (MCAO) for 45 min, followed by 7 days of reperfusion. The in vitro study demonstrated that SCE protected cells against etoposide-induced cell viability loss in SH-SY5Y cells. Apoptotic phenotypes, such as cleaved PARP and caspase-3, and oxidative stress in etoposide-treated cells were ameliorated by SCE treatment. In MCAO-reperfusion injury, SCE promoted neuronal survival and level of brain-derived neurotrophic factor (BDNF) by reducing glial activation, oxidative stress, and apoptosis in the ipsilateral cortex. These results indicated that SCE exerted protective effects under etoposide treatment and in a MCAO-reperfusion model by reducing JNK and p38 MAPK activation. This study presents the first evidence that SCE has therapeutic potential for the treatment of ischemic stroke or neurological disorder-related cell death.

Measuring levels of biogenic amines and their metabolites in rat brain tissue using high-performance liquid chromatography with photodiode array detection.

We developed a method to detect biogenic amines and their metabolites in rat brain tissue using simultaneous high-performance liquid chromatography and a photodiode array detection. Measurements were made using a Hypersil Gold C-18 column (250 × 2.1 mm, 5 µm). The mobile phase was 5 mM perchloric acid containing 5 % acetonitrile. The correlation coefficient was 0.9995-0.9999. LODs (S/N = 3) and LOQs (S/N = 10) were as follows: dopamine 0.4 and 1.3 pg, 3, 4-dihydroxyphenylacetic acid 8.4 and 28.0 pg, serotonin 0.4 and 1.3 pg, 5-hydroxyindolacetic acid 3.4 and 11.3 pg, and homovanillic acid 8.4 and 28.0 pg. This method does not require derivatization steps, and is more sensitive than the widely used HPLC-UV method.

Ethanol extracts of Sanguisorba officinalis L. suppress TNF-α/IFN-γ-induced pro-inflammatory chemokine production in HaCaT cells.

BACKGROUND: Sanguisorba officinalis L. (SOL) is a perennial plant widely distributed in Asia, its roots are well-known as a traditional herbal medicine to treat burns, chronic intestinal infections, scalds, and inflammation in Korea. Also, the roots of SOL are used for treatment of many types of allergic skin diseases, including urticarial, eczema, and allergic dermatitis. PURPOSE: In this study we investigated the underlying mechanism of anti-inflammatory effect of an ethanol extract of SOL roots (ESOL). STUDY DESIGN: The ability of ESOL to inhibit inflammatory skin disorder was tested in human keratinocyte HaCaT cells. METHODS: Viability test using MTT assay were used to determine non-cytotoxic concentrations of ESOL on HaCaT cells. ESOL-mediated inhibition of the tumor necrosis factor (TNF)-α/interferon (IFN)-γ-induced production of pro-inflammatory chemokines-such as macrophage-derived chemokine (MDC), regulated on activation, normal T-cell expressed and secreted (RANTES), interleukin (IL)-8, and thymus and activation regulated chemokine (TARC)-at the mRNA level was determined by real-time reverse transcription-polymerase chain reaction (RT-PCR). The ability of ESOL to reduce the expression of pro-inflammatory marker proteins was evaluated by Western blot analysis and immunocytochemistry. RESULTS: ESOL reduced the production of MDC, RANTES, IL-8, and TARC in HaCaT cells stimulated with TNF-α/IFN-γ at both protein and mRNA levels. ESOL also suppressed the phosphorylation of signal transducer and activator of transcription (STAT)-1, extracellular signal-regulated kinase (ERK), and inhibited both nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor-alpha (IκB-α) degradation and the nuclear translocation of NF-κB/p65. ESOL exerts anti-inflammatory effects by suppressing the expression of TNF-α/IFN-γ-stimulated chemokines and pro-inflammatory molecules via a blockade NF-κB, STAT-1, and ERK activation. CONCLUSION: Our results suggest the preventive potential of ESOL as a herbal medicine for the treatment of inflammatory skin diseases.

Inhibitory effects of palmultang on inflammatory mediator production related to suppression of NF-κB and MAPK pathways and induction of HO-1 expression in macrophages.

Palmultang (PM) is an herbal decoction that has been used to treat anorexia, anemia, general prostration, and weakness due to chronic illness since medieval times in Korea, China, and Japan. The present study focused on the inhibitory effects of PM on the production of inflammatory factors and on the activation of mechanisms in murine macrophages. PM suppressed the expression of nitric oxide (NO), inflammatory cytokines and inflammatory proteins by inhibiting nuclear factor (NF)-κB and mitogen-activated protein kinase (MAPK) signaling pathways and by inducing heme oxygenase (HO)-1 expression. Collectively, our results explain the anti-inflammatory effect and inhibitory mechanism of PM in macrophages stimulated with lipopolysaccharide (LPS).

Galla rhois exerts its antiplatelet effect by suppressing ERK1/2 and PLCß phosphorylation.

Galla rhois and its components have various biological activities, including protective effects on liver cells as well as antimetastatic, antiplatelet, and antibacterial effects. In the present study, we identified the antiplatelet activity and possible mechanism of action of a G. rhois extract (GRE). We investigated the effect of GRE and its components on rabbit platelet activation, and their possible molecular mechanisms. The GRE inhibited collagen-, AA-, and thrombin-induced platelet aggregation as well as serotonin secretion, in a concentration-dependent manner. The GRE significantly inhibited the production of lipoxygenase-mediated 12-hydroxyeicosatetraenoic acid. The GRE effectively suppressed thrombin-stimulated PLCß3 phosphorylation and collagen-induced ERK1/2 phosphorylation, in addition, the GRE significantly restored the cAMP level, which had decreased due to collagen or thrombin. Among the components of GRE, methyl gallate inhibited the collagen-induced platelet activation through suppression of ERK phosphorylation, penta-O-galloyl-ß-D-glucoside inhibited the thrombin-induced platelet activation through suppression of PLCß phosphorylation. These results indicate that the GRE including methyl gallate and penta-O-galloyl-ß-D-glucoside suppressed platelet activation by inhibiting ERK1/2 and PLCß3 phosphorylation.

Survival of porcine fibroblasts enhanced by human FasL and dexamethasone-treated human dendritic cells in vitro.

Cell-mediated and acute vascular rejections remain to be one of the primary hurdles to achieve successful xenotransplantation. Fas ligand is known to be an important molecule for the formation of 'immune-privileged' condition and dendritic cells treated with dexamethasone (Dex-DCs) acting like tolerogenic DCs (tDCs) which are known to protect transplanted cells and organs from unwanted immune responses. The present study investigated the possibility that porcine fibroblasts expressing human Fas ligand (PhF) together with human Dex-DCs could induce prolonged survival of porcine fibroblasts in vitro. PhF was collected from an ear of human Fas ligand transgenic porcine and cell-line was established by MGEM Inc. PhF labeled with CFSE co-cultured with human peripheral blood mononuclear cells (hPBMCs) were examined with respect to induction of tolerance and cell death when co-cultured with Dex-DCs for 3days. PhF induced the apoptosis in hPBMCs, especially CD4(+) T cells. Dex-DCs showed significant (P<0.05) reduction on the expression of CD80, CD86 and MHC class I/II, and the secretion of IL-12p70, TNF-α and IL-10, but increase of latency-associated peptide (LAP). Survival of PhF was significantly higher than that of WT and it was increased in the presence of Dex-DCs when compared to the other DCs (i.e.,DCs, LPS-treated DCs and LPS/Dex-treated DCs) in vitro. Survival of PhF did not change by co-culture with Dex-DCs due to apoptotic cell death of Dex-DCs. Dex-DCs reduced the death of porcine fibroblasts and, at the same time, PhF induced the apoptosis from hPBMCs, but it was not synergistic.

Vitex rotundifolia L. prevented airway eosinophilic inflammation and airway remodeling in an ovalbumin-induced asthma mouse model.

Vitex rotundifolia L. (VR) as long been used in China and Korea in traditional medicine. This study was conducted to evaluate the ability of Vitex rotundifolia L. to prevent airway inflammation and remodeling in an ovalbumin (OVA)-induced murine asthma model. The total cell number and number of inflammatory cells in the bronchoalveolar lavage (BAL) fluid were counted. The levels of cytokines in the BAL fluid and serum IgE levels were measured using an ELISA. For histological analysis, hematoxylin and eosin staining, periodic acid-Schiff staining and immunohistochemistry were evaluated. The release of total cells into the BAL fluid was significantly inhibited in OVA-induced asthmatic mice treated with VR extract. In addition, eosinophilia and lymphocytosis were reduced significantly in mice that received VR extract. Furthermore, levels of the T(h)2 cytokines IL-4 and IL-5 and pro-inflammatory cytokine TNF-α in the BAL fluid and total IgE in serum were markedly suppressed by VR extract. OVA-specific IgE in the serum and IL-13 in the BAL fluid were decreased, but not significantly. The allergic effects of VR extract were accompanied by a reduction in airway hyperresponsiveness. Additionally, morphologic findings demonstrated that VR extract substantially inhibited OVA-induced eosinophilia, goblet cell hyperplasia and smooth muscle mass production. This finding suggests that VR extract may have pharmacological effects that would be useful for the treatment of asthma via the inhibition of the T(h)2 response and airway remodeling.

Analysis of methanogenic archaeal communities of rumen fluid and rumen particles from Korean black goats.

Molecular diversity of methanogens in the rumen of Korean black goats was investigated with 16S rRNA gene clone libraries using methanogen-specific primers. The libraries were composed of rumen fluid-associated methanogens (FAM) and rumen particle-associated methanogens (PAM) from rumen-fistulated Korean black goats. Among the 141 clones of the FAM library, the sequences were mostly related to two phyla, the Methanobacteriaceae family (77.3%) and the Thermoplasmatales family (22.7%); and among the 68 clones of the PAM library, sequences were also mainly clustered in the two phyla, the Thermoplasmatales family (63.24%) and the Methanobacteriaceae family (35.29%). Most of the sequenced clones in the two libraries were closely related to uncultured methanogenic archaeon. Quantitative real-time PCR revealed that PAM (8.97 log 10) had significantly higher (P < 0.01) density of methanogens by the methanogenic 16S rRNA gene copies than FAM (7.57 log 10). The two clone libraries also showed difference in Shannon index (FAM library 1.70 and PAM library 1.59) and Chao 1 estimator (FAM library 18 and PAM library 17 operational taxonomic units). Apparent differences found in the microbial community from the two 16S rRNA gene libraries could be a result of such factors as the chemical and physical nature of the target material surface, types or component of diets, the interaction between the methanogens and other microbes, and age of the experimental goats.