Faculty of Science - Mahidol University

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324 The water decoction of heartwood and stem bark of Ventilago harmandiana Pierre (Family Rhammnaceae) are used in Thai traditional medicine for the treatment of diabetes as well as wound and chronic inflammation. Methanolic extracts from the heart wood, stem bark, stem wood and pure compounds of the herb exerted strong inhibitory effects on the acute phase of inflammation as seen in ethyl phenylpropiolate (EPP)-induced ear edema as well as in carragenin-induced paw edema in rats. Activated macrophages participate actively in the inflammatory response by producing large amounts of pro-inflammatory cytokines such as TNF-α, chemoattractant cytokines such as IL-8 and pro-inflammatory products of arachidonic acid metabolism as well as nitric oxide. This study examined whether a quinone and two anthraquinone compounds from Ventilago harmandiana modulate the production of PGE 2 , tumour necrosis factor-á (TNF-á) and nitric oxide (NO) by LPS-activated human macrophages. The expression of the mRNA of COX-2 and TNF-á was also investgated. All three compounds from Ventilago harmandiana were shown to significantly inhibit the production of TNF-á and PGE 2 in a concentration-dependent manner without notable cytotoxic effects on these cells. The expression of both TNF-á and COX-2 mRNA was also significantly inhibited by these compounds. However, neither the production of NO nor iNOS mRNA expression could be detected from LPS-human activated macrophages. These results indicated that the inhibitory effects of a quinone and the two anthraquinone compounds from Ventilago harmandiana Pierre on the production, by activated human macrophages, of PGE 2 and TNF-á which is regulated at the transcriptional level might be attributed, in part, to their antiinflammatory activities. INHIBITORY EFFECTS OF NEUTROPHIL FUNC- TION AND T-LYMPHOCYTES PROLIFERTION BY PUERARIA MIRIFICA EXTRACTS (NO. 856) Payong Wanikiat 1 , Yupin Sanvarinda 1 , Taworn Jaipetch 2 , Vichai Reutrakul 2 1 Department of Pharmacology, 2 Department of Chemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand. Key words : Pueraria mirifica; Inflammation; Anti-inflammation; Neutrophil functions; Chemotaxis; Superoxide anion generation; MPO production; Elastase release Pueraria mirifica (P. mirifica) Airy Shaw (Legumunosae), a phytoestrogen – rich plant with a local name of white Kwao Krrua, has long been used in Thailand as a rejuvenating folk medicine. It has also been used by menopausal women for the hormonal replacement purpose. The chemical compounds found in the plant including Isoflavones and glycosides e.g. daidzein, genistein, kwakurin; Chromenes e.g. miroestrol, deoxymiroestrol, isomiroestrol, Comestans e.g. coumestrol and Pterocarpans e.g. puemiricarpene etc. It is believed that the content of phytoestrogen in the plants are of therapeutic benefit. Some active ingredients of P. mirifica such as miroestrol exhibit estrogenic like activity. Estrogen plays a protective role on cardiovascular system and has been known to exert anti-inflammatory effects both in vivo and in vitro by inhibiting the expression and production of various proinflammatory mediators. In this study, methanol and ethylacetate extracts of Pueraria mirifica were investigated for their antiinflammatory activities based on neutrophil functions, including chemotaxis, superoxide anion generation (SAG), myeloperoxidase (MPO) production, and elastase release, and on T-lymphocyte proliferation. Both crude extracts were primarily investigated for their cytotoxic effects on both neutrophils and lymphocytes using MTT assay. Neutrophil viability was not affected by these compounds (1-100 μg/ml) either at 4 hours incubation period, except at a concentration of 1000 μg/ml (IC 50 >1000). After incubation of human neutrophils with various concentrations of both methanol and ethylacetate extracts of Pueraria mirifica, neutrophil chemotaxis, SAG, MPO production, and elastase release induced by N-formyl-methionyl-leucyl-phenylalanine (fMLP) were strongly inhibited in a concentration-dependent manner. Both crude extracts also inhibited T-lymphocyte proliferation as quantified by [H 3 ] thymidine incorporation. These results suggested that the inhibitory effects of the Pueraria mirifica extracts on neutrophil functions and T-lymphocyte proliferation might be attributed, in part, to their anti-inflammatory activities. CONCERTED MECHANISM OF SWE1/WEE1 REGULATION BY MULTIPLE KINASES IN BUDDING YEAST. (NO. 857) Satoshi Asano 1 , Jung-Eun Park 1 , Krisada Sakchaisri 1, 2 , Li- Rong Yu 3 , Sukgil Song 4 , Porntip Supavilai 2 , Timothy D Veenstra 3 and Kyung S Lee 1 1 Laboratory of Metabolism, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, USA. 2 Department of Pharmacology, Faculty of Science, Mahidol University, Bangkok, Thailand. 3 Laboratory of Proteomics and Analytical Technologies, NCI-Frederick, Frederick, MD, USA. 4 College of Pharmacy, Chungbuk National University, South Korea. Key words : Cdc5, Cdc28, G2/M transition, Swe1 In eukaryotes, entry into mitosis is induced by cyclin Bbound Cdk1, which is held in check by the protein kinase, Wee1. In budding yeast, Swe1 (Wee1 ortholog) is targeted to the bud neck through Hsl1 (Nim1-related kinase) and its adaptor Hsl7, and is hyperphosphorylated prior to ubiquitin-mediated degradation. Here, we show that Hsl1 and Hsl7 are required for proper localization of Cdc5 (Polo-like kinase homolog) to the bud neck and Cdc5dependent Swe1 phosphorylation. Mitotic cyclin (Clb2)-bound Cdc28 (Cdk1 homolog) directly phosphorylated Swe1 and this modification served as a priming step to promote subsequent Cdc5dependent Swe1 hyperphosphorylation and degradation. Clb2-Cdc28 also facilitated Cdc5 localization to the bud neck through the enhanced interaction between the Clb2-Cdc28-phosphorylated Swe1 and the polo-box domain of Cdc5. We propose that the concerted action of Cdc28/Cdk1 and Cdc5/Polo on their common substrates is an evolutionarily conserved mechanism that is crucial for effectively triggering mitotic entry and other critical mitotic events. (EMBO J. 24, (2005) 2194-2204.) PDF created with FinePrint pdfFactory Pro trial version http://www.pdffactory.com Faculty of Science
Mahidol University Annual Research Abstracts, Vol. 33 325 INTERLEUKIN-5 REDUCES THE EXPRESSION OF UTEROGLOBIN-RELATED PROTEIN (UGRP) 1 GENE IN ALLERGIC AIRWAY INFLAMMATION (NO. 858) Yoshihiko Chiba 1, 2 , Achara Srisodsai 1, 3 , Porntip Supavilai 3 and Shioko Kimura 1 1 Laboratory of Metabolism, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA 2 Department of Pharmacology, Hoshi University, Tokyo 142-8501, Japan 3 Department of Pharmacology, Faculty of Science, Mahidol University, Bangkok 10400, Thailand. Key words : Uteroglobin-related protein 1 (UGRP 1), Interleukin- 5 (IL-5), Allergic asthma Airway inflammation is thought to play a major role in the pathogenesis of bronchial asthma. The precise role of individual inflammatory cells, mediator and asthma related genes in allergic lung diseases is not completely understood. The uteroglobin-related protein (UGRP) 1 was proposed to be an asthma candidate gene and play a role in regulating lung inflammation, however its precise function in the airways remains obscure. In this investigation, we used a mouse model of allergic airway inflammation to establish a relationship between UGRP 1 and IL-5 in airway inflammation. Ovalbumin (OVA) challenged mice demonstrate eosinophilia in airway tissues and high levels of IL-5 in bronchoalveolar lavage (BAL) fluid analogous to that found in bronchial asthma. Interestingly, these “OVA-challenged” mice show down-regulation of Ugrp1 expression as compared with the control group. Regression analysis further demonstrates a significant negative correlation between Ugrp1 mRNA expression in the lung and IL-5 levels in BAL fluid with r = 0.948 and P < 0.0001 when IL-5 levels were normalized by log transformation. Intranasal instillation of IL-5 to mice revealed an inhibitory effect of IL-5 on the expression of Ugrp1 mRNA. Together, these results indicate an involvement of IL-5 in the down-regulation of Ugrp1 expression in airway inflammation such as allergic asthma disease. (Immunol Lett. 97(2005)123-129.) ANTIOXIDANT EFFECTS OF AQUEOUS EXTRACTS FROM DRIED CALYX OF HIBISCUS SABDARIFFA LINN. (ROSELLE) IN VITRO USING RAT LOW- DENSITY LIPOPROTEIN (LDL) (NO. 859) Vilasinee Hirunpanich 1 , Anocha Utaipat 1 , Noppawan Phumala Morales 2 , Nuntavan Bunyapraphatsara 3 , Hitoshi Sato 4 , Angkana Herunsalee 5 and Chuthamanee Suthisisang 1 1 Department of Pharmacology, Faculty of Pharmacy, Mahidol University; 2 Department of Pharmacology, Faculty of Science, Mahidol University; 3 Department of Pharmacognosy, Faculty of Pharmacy, Mahidol University; 4 Department of Clinical and Molecular Pharmacokinetics/Pharmacodynamics, Faculty of Pharmaceutical Science, Showa University; 5 Medicinal Plant Research Institute Department of Medical Sciences, Ministry of Public Health, Thailand. Key words: Hibiscus sabdariffa; roselle; low-density lipoprotein (LDL) oxidation; antioxidant The present study quantitatively investigated the antioxidant effects of the aqueous extracts from dried calyx of Hibiscus sabdariffa LINN. (roselle) in vitro using rat low-density lipoprotein (LDL). Formations of the conjugated dienes and thiobarbituric acid reactive substances (TBARs) were monitored as markers of the early and later stages of the oxidation of LDL, respectively. Thus, we demonstrated that the dried calyx extracts of roselle exhibits strong antioxidant activity in Cu2+-mediated oxidation of LDL (p
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