Anti-neuroinflammatory Effects of Ethanol Extract of Inula helenium L (Compositae)

Purpose: To evaluate the in vitro antioxidant and anti- neuroinflammatory activities of Inula helenium extract (IHE) against lipopolysaccharide (LPS)-induced production of nitric oxide (NO) by primary microglial cells.Methods: Cell viability was estimated by 3-(4, 5-dimethylthiazol-2- yl)-2, 5- diphenyl-tetrazolium bromide (MTT) assay. Antioxidant activity was evaluated using 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging assay. LPS-stimulated BV-2 microglia were used to study the expression and production of inflammatory mediators, including NO, inducible NO synthase (iNOS)and Interukin-6 (IL-6).Results: Pretreatment with IHE prior to LPS treatment significantly inhibited excessive production of NO (p < 0.001 at 20, 40, 80 and 100 μg/mL) in a dose-dependent manner, and was associated with downregulation of expression of inducible nitric oxide synthase (iNOS). IHE also suppressed the LPSinduced increase in IL-6 level (p < 0.01 at 40 and 80 μg/mL) in BV-2 cells. The antioxidant activity exhibited by IHE might play a critical role in ameliorating neuroinflammatory processes in LPSstimulated BV-2 microglial cells.Conclusion: IHE may be beneficial in preventing and treating neurodegenerative and oxidative stressrelated diseases.Keywords: Inula helenium, DPPH, Neurodegenerative diseases, Anti inflammatory, Anti-oxidant, Microglial cel

Cytotoxic T-lymphocyte Antigen-4 Binding to SHP2 Interacting Transmembrane Adapter Protein by Phosphorylation in T-Cell

Purpose: To investigate potential cytotoxic T-lymphocyte antigen-4 (CTLA-4) binding partners and assess whether potential binding partners affect the full function of CTLA-4. .Methods: The down-regulation effects of CTLA-4 and SIT were assessed by culturing CD3 stimulated T-cells. CTLA-4 and SIT proteins were measured by immunoblot analysis and production of interlukin-2 transcription activity evaluated by luciferase assay.Results: CTLA-4 inhibited the interlukin-2 production capacity of CD3-stimulated T cells. CTLA-4 interaction with SHP2 interacting transmembrane adapter protein (SIT) in the down-regulation of the transcription of Interulin-2 required CTLA-4 binding to SIT tyrosine motifs. The SIT tyrosine mutants were significantly lower (25 – 75 %) after phosphorylation compared with WT-SIT (transfected cells, p < 0.05) and untreated control. The remaining 90 % phosphorylation in the F188ANS mutant can be explained by phosphorylation of other tyrosines in the sequence of SIT (p < 0.05). For interukin-2 transcription, F188ANS single mutant and double F148SEV mutant, increased NF-AT activity by 35 % compared with the wild type (p < 0.05).Conclusion: The findings imply that SIT transmembrane adaptor (SIT) protein, binds to CTLA-4 and thus potentiates the inhibitory role of this co-receptor. This phenomenon may lead to the development of new treatment strategies for autoimmune diseases and graft rejection.Keywords: Cytotoxic T-lymphocyte antigen-4, Interleukin-2, Nuclear factor of activated T-cells/Activator protein-1, SHP2 interacting transmembrane adapter protein, Autoimmune diseases, Graft rejectio

A carbon CT system: how to obtain accurate stopping power ratio using a Bragg peak reduction technique

学位記番号：医博甲1688

Ameliorative effect of Houttuynia cordata Thunb (Saururaceae) leaf extract in loperamide-induced constipation in rats

Purpose: To investigate the efficacy of Houttuynia cordata Thunb (Saururaceae) extract against loperamide-induced constipation in Wistar rats. Methods: Constipation was induced by loperamide (3 mg/kg). The rats were orally treated daily with either 40, 80, 160 mg/kg/day of H. cordata extract or 0.25 mg/kg bisacodyl (positive control) for 7 days, while normal control group received water only. Feed and water intake, body weight, number and water content of fecal pellets were monitored throughout the study period. The biochemical marker activities of aspartate aminotransferase (AST), γ-glutamyltransferase (γ-GT) and lactate dehydrogenase (LDH) were determined in the blood of the animals. Results: The water content of fecal pellets and body weight gain of the constipation rats changed significantly (p < 0.05) when compared to the normal control. Serum levels of AST were significantly higher (p < 0.05) in the 80 and 160 mg/kg dose groups when compared to normal controls. However, there were no alterations in the levels of γ-GT and LDH across all groups. Conclusion: Treatment of rats with ethanol extract of H. cordata leaf has useful effects against constipation. Therefore, H. cordata leaf may be benefitial as a traditional herbal medicine for the management of constipation

Saussurea lappa Clarke extract exhibits potent antioxidant effect and attenuates neuroinflammatory responses in lipopolysaccharide-stimulated microglial cells

Purpose: To investigate the antioxidant and anti-neuroinflammatory potential of Saussurea lappa Clarke (SLC-EA) extract in LPS-stimulated BV-2 microglial cells.Methods: Cell viability was measured by using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay while antioxidant activity was evaluated by using the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity. Lipopolysaccharide (LPS) was used to stimulate BV-2 microglia. Griess assay was employed to assess nitric oxide (NO) production. iNOS (inducible NO synthase) expression and TNF-α (tumor necrosis factor-alpha) cytokine production were measured by ELISA (enzyme-linked immunosorbent assay) and immuno blot analysis, respectively.Results: Pretreatment of 100 mg/ml of SLC-EA (p < 0.001) was inhibited Nitric Oxide (NO) by 1 ug/ml of LPS-treated murine BV-2 cells. The expression of iNOS and TNF-α were reduced by SLC-EA concentration dependent manner (p < 0.001 at 100 mg/ml). SLC-EA were scavenged 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radicals in a dose-dependent manner with an IC50 value of approximately 51.4 μg/ml.Conclusion: The results indicate that SLC-EA extract exhibits strong antioxidant properties and inhibits excessive pro-inflammatory cytokine due probably to the antioxidant phenolic compounds present in SLC-EA extract. Further work in exploring the in-depth mechanisms of SLC-EA extract in regulating inflammatory signaling pathways in treating neuroinflammatory diseases is necessary. Keywords: Saussurea lappa, Antioxidant, Neuroinflammation, Microglia, TNF-α, iNO

Evaluation of antioxidant and anti-inflammatory effects of three different Rubus coreanus Miq. by-products

Purpose: To investigate the antioxidant and anti-inflammatory properties of three different Rubus coreanus Miq. by-products in stimulated BV-2 microglial cells and explore its underlying physiological efficacy.Methods: Cell viability assessment was performed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Lipopolysaccharide (LPS) was used to activate BV-2microglia. Total polyphenol and total flavonoid contents were determined by the method of Folin-Denis. As three different Rubus coreanus Miq. by-products remaining after extraction of Rubus coreanus, High performance liquid chromatography (HPLC)) finger printing, ABST (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging assay, and NO (nitric oxide) inhibitory assay were performed.Results: Three different Rubus coreanus by-product extract did not exhibit any signs of cytotoxicity to BV-2 cells up to 100 μg/ml concentration (p < 0.5). The LPS-activated excessive release of NO in BV-2 cells was significantly inhibited by Rubus coreanus by-product extract (p < 0.5) at 500 μg/mL). Total polyphenol and total flavonoid contents were highest in 50 % ethanol wine processing by-product (p < 0.5 at 30, 50, 70 and 100 %, respectively). The by-product of wine processing had the lowest RC50 radical scavenging effect (16.53 μL/ml). The quercetin content of the wine processing by-product was the highest in the 70% ethanol extract at 6.26 mg/g (p < 0.5 at 30, 50, 70 and 100%, respectively).Conclusion: These results reveal that of the three other by-products, wine processing by-product has the highest antioxidant and anti-inflammatory activities. The use of these by-products has high added value for industrial production; furthermore, they are a potential treatment for various inflammatory diseases

Reliability of DEXA on Body Composition in Korean Athletes

PURPOSE: The purpose of this study was to assess the reliability of DEXA for measuring body composition in Korean Athletes. METHODS: Twenty-nine athletes (n=29) registered for the college athlete program voluntarily participated in the study. Participants’ height and weight were measured, and BMI (Body Mass Index) was calculated before the participants’ body composition was measured. Muscle mass (kg), lean mass (kg), bone mineral density (BMC) (g·cm-2), and total fat mass (kg) of each participant was assessed by DEXA lunar DPX-L (GE Lunar, Madison, USA) for four times within a day to examine the difference by time frames. Four trials consist of ‘early in the morning × 2 with fasting’ with 30min break between two trials, ‘after lunch × 2’ with 30 min break between the two trials. Intra-class correlation (ICC) was conducted for overall reliability (p\u3c0.05) and a repeated measure ANOVA was performed to compare the difference of each trial (p\u3c0.05). RESULTS: The mean ± SD of muscle mass, lean mass, BMC, and fat mass was 56.4 ± 4.6kg, 59.4 ± 5.0kg, 2.3 ± 0.4g·cm-2, and 9.3 ± 4.8kg respectively. Each trail (mean ± SD) of muscle mass were 56.4 ± 4.7kg, 56.1 ± 4.8kg, 56.5 ± 4.6kg, and 56.4 ± 4.7kg, respectively, lean mass were 59.4 ± 5.1kg, 59.2 ± 5.1kg, 59.5 ± 5.0kg, and 59.4 ± 5.0kg, respectively, BMC were 3.0 ± 0.4g·cm-2, 3.0 ± 0.4g·cm-2, 3.0 ± 0.4g·cm- 2, and 3.0 ± 0.4g·cm-2, respectively, and fat mass were 9.3 ± 4.9kg, 9.2 ± 4.8kg, 9.3 ± 4.9kg, and 9.3 ± 4.9kg, respectively. Reliability of the ICC test showed strong agreement on muscle mass (r=0. 994 and p\u3c0.0001), lean mass (r=0. 995 and p\u3c0.0001), BMC (r=0. 995 and p\u3c0.0001), and fat mass (r=0. 998 and p\u3c0.0001). Cronbach’s alpha were 0.99 (muscle mass), 0.99 (Lean Mass), 0.99 (BMC), and 1.00 (Fat mass). No significant difference between each trial was observed in fat mass (p\u3e0.36). However, there was a significant difference in muscle mass (p\u3c0.001), lean mass (p\u3c0.001), and BMC (p\u3c0.04). CONCLUSION: Although all of the variables showed strong agreement on overall reliability from the ICC test, the reliability for the muscle mass, lean mass, and BMC showed significant differences in different time frame