Antitumor effects of SB injection in canine osteosarcoma and melanoma cell lines.

The present study was designed to evaluate the effect of SB injection, which is composed of extracts from the roots of Pulsatilla koreana, Panax ginseng, and Glycyrrhiza glabra, on the viability of canine osteosarcoma and melanoma cells and nonneoplastic canine cells. Cells were treated with SB injection, conventional chemotherapeutic drugs, or a combination of both at various concentrations. Cellular viability was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Flow cytometry was used to evaluate the cell cycle and apoptosis. SB injection inhibited the growth of osteosarcoma and melanoma cells in a dose-dependent manner. The cell cycle of the affected cells was arrested in the G2/M phase, indicating an anti-proliferative effect. SB injection dose-dependently increased the rate of apoptosis. Furthermore, we found that combining SB injection with chemotherapeutic drugs resulted in a greater reduction in canine malignant cell proliferation than either treatment alone. SB injection did not affect the viability of peripheral blood mononuclear cells regardless of concentration, which suggested that SB injection did not suppress the activity of normal cells. This study suggested that SB injection can be considered an effective alternative medication for animal cancers in veterinary medicine.

Electroacupuncture at the Zusanli and Baihui acupoints ameliorates type-2 diabetes-induced reductions in proliferating cells and differentiated neuroblasts in the hippocampal dentate gyrus with increasing brain-derived neurotrophic factor levels.

In the current study, we investigated whether electroacupuncture (EA) can inhibit pathological reductions in neurogenesis. Zucker diabetic fatty (ZDF) rats at 7 weeks of age were anesthetized with zoletil, and sham-acupuncture or EA at the Zusanli (ST36) and Baihui (GV20) acupoints was administered once a day for 5 weeks. In the ZDF group that received sham-EA (ZDF-Sham group), the blood glucose level was significantly increased together with age as compared to the control littermates [Zucker lean control (ZLC) rat]. In contrast, proliferating cells and differentiated neuroblasts were significantly decreased in the ZDF-Sham group compared to the ZLC group. Although EA treatment decreased blood glucose levels, this was not statistically significant when compared to blood glucose levels changes in the ZDF-Sham group. However, proliferating cells and differentiated neuroblasts were significantly increased with EA in ZDF rats as compared to those in the ZDF-Sham group. Brain-derived neurotrophic factor (BDNF) levels were significantly decreased in hippocampal homogenates of ZDF-Sham group compared to those in the ZLC group. The EA treatment significantly increased the BDNF levels compared to those in the ZDF-Sham group, and BDNF levels in this group were similar to those in the ZLC group. These results suggest that EA at ST36 and GV20 can ameliorate the reductions in proliferating cells and differentiated neuroblasts in the dentate gyrus induced by type-2 diabetes without significantly reducing blood glucose levels with increasing BDNF levels.

Anti-inflammatory effects of ethanolic extract from Sargassum horneri (Turner) C. Agardh on lipopolysaccharide-stimulated macrophage activation via NF-κB pathway regulation.

Inflammation is major symptom of the innate immune response by infection of microbes. Macrophages, one of immune response related cells, play a role in inflammatory response. Recent studies reported that various natural products can regulate the activation of immune cells such as macrophage. Sargassum horneri (Turner) C. Agardh is one of brown algae. Recently, various seaweeds including brown algae have antioxidant and anti-inflammatory effects. However, anti-inflammatory effects of Sargassum horneri (Turner) C. Agardh are still unknown. In this study, we investigated anti-inflammatory effects of ethanolic extract of Sargassum horneri (Turner) C. Agardh (ESH) on RAW 264.7 murine macrophage cell line. The ESH was extracted from dried Sargassum horneri (Turner) C. Agardh with 70% ethanol and then lyophilized at -40 °C. ESH was not cytotoxic to RAW 264.7, and nitric oxide (NO) production induced by LPS-stimulated macrophage activation was significantly decreased by the addition of 200 µg/mL of ESH. Moreover, ESH treatment reduced mRNA level of cytokines, including IL-1ß, and pro-inflammatory genes such as iNOS and COX-2 in LPS-stimulated macrophage activation in a dose-dependent manner. ESH was found to elicit anti-inflammatory effects by inhibiting ERK, p-p38 and NF-κB phosphorylation. In addition, ESH inhibited the release of IL-1ß in LPS-stimulated macrophages. These results suggest that ESH elicits anti-inflammatory effects on LPS-stimulated macrophage activation via the inhibition of ERK, p-p38, NF-κB, and pro-inflammatory gene expression.

Genistein inhibits pro­inflammatory cytokines in human mast cell activation through the inhibition of the ERK pathway.

Anaphylaxis is a rapidly occurring allergic reaction to any foreign substance, including venom from insects, foods and medications, which may cause fatalities. To prevent anaphylaxis, these triggers must be avoided. However, avoidance of numerous triggers is difficult. For this reason, the development of immunotherapeutic adjuvants that suppress the allergic response is important for anaphylaxis control. Mast cells are one of the major inflammatory cells involved in the inflammatory response, which secrete several inflammatory cytokines, including tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1ß, and recruits other immune cells. Mast cells are also involved in a number of diseases, such as sinusitis, rheumatoid arthritis and asthma. Genistein, a phytoestrogen, has been reported to have anti-oxidative and anti-inflammatory activities. However, the effects of genistein on the anti-inflammatory response of mast cells remain unknown. In the present study, the anti-inflammatory effects of genistein on mast cells were investigated. Genistein significantly decreased IL-6 and IL-1ß mRNA levels, as well as IL-6 production in PMA/A23187-induced mast cells activation. In addition, genistein inhibited the phosphorylation of ERK 1/2 in PMA/A23187-induced mast cell activation. However, phosphorylation of p38 was not altered. Thus, these findings indicate that genistein inhibited the inflammatory status of mast cells through inhibition of the ERK pathway.

Effects of electroacupuncture at Zusanli and Baihui on brain-derived neurotrophic factor and cyclic AMP response element-binding protein in the hippocampal dentate gyrus.

Previously, we observed that electroacupuncture (EA) at ST36 (Zusanli) and GV20 (Baihui) enhanced cell proliferation and neuroblast differentiation in the rat dentate gyrus. In this study, we investigated the possible mechanisms of EA in this effect. For this, we applied EA at ST36 and GV20 of Wistar rats (13-week-old) once a day for 3 weeks. Application of EA at these acupoints significantly increased the number of phosphorylated cyclic AMP response element-binding protein (pCREB)-immunoreactive cells in the dentate gyrus. In addition, EA significantly increased the levels of brain-derived neurotrophic factor (BDNF) and pCREB protein in the dentate gyrus. The administration of K252a, an inhibitor of BDNF receptor, significantly reduced cell proliferation in the subgranular zone of dentate gyrus. These results suggest that EA significantly increased neuroblast plasticity via pCREB and BDNF activation in the dentate gyrus.

Comparing the effects of acupuncture and electroacupuncture at Zusanli and Baihui on cell proliferation and neuroblast differentiation in the rat hippocampus.

We compared the effects of acupuncture and electroacupuncture on cell proliferation and neuroblast differentiation using specific markers, Ki67 and doublecortin (DCX), in the subgranular zone of the dentate gyrus (SZDG) in 13-week old Wistar rats. Acupuncture and electroacupuncture were applied simultaneously in the acu-points, ST36 (Zusanli) and GV20 (Baihui), once a day for 3 weeks. Acupuncture and electroacupuncture at these acu-points significantly increased the number of Ki67-positive cells and DCX-immunoreactive neuroblasts compared to the control or sham acupuncture group. Electroacupuncture treatment significantly increased the number of well-developed (tertiary) dendrites in the SZDG compared to acupuncture treatment. These results suggest that both acupuncture and electroacupuncture increase neurogenesis in the normal, but that electroacupuncture has greater effects on neuroblast plasticity than acupuncture in the dentate gyrus.