Antiplatelet effect of sanguinarine is correlated to calcium mobilization, thromboxane and cAMP production.

Sanguinarine is a plant alkaloid present in the root of Sanguinaria canadensis and Poppy fumaria species. Sanguinarine has been used as an antiseptic mouth rinse and a toothpaste additive to reduce dental plaque and gingival inflammation. In this study, we investigated the antiplatelet effects of sanguinarine, aiming to extend its potential pharmacological applications. Sanguinarine inhibited platelet aggregation induced by arachidonic acid (AA), collagen, U46619 and sub-threshold concentration of thrombin (0.05 U/ml) with IC(50) concentrations of 8.3, 7.7, 8.6 and 4.4 microM, respectively. Sanguinarine (5-10 microM) inhibited 10-31% of platelet TXB(2) production, but not platelet aggregation induced by higher concentration of thrombin (0.1 U/ml). SQ29548, a thromboxane receptor antagonist, inhibited the AA-induced platelet aggregation but not TXB(2) production. Sanguinarine suppressed cyclooxygenase-1 (COX-1) activity (IC(50)=28 microM), whereas its effect on COX-2 activity was minimal. Sanguinarine (8, 10 microM) further inhibited the AA-induced Ca(2+) mobilization by 27-62%. In addition, SQ22536, an adenylate cyclase inhibitor, attenuated the inhibitory effect of sanguinarine toward AA-induced platelet Ca(2+) mobilization and aggregation. These results suggest that sanguinarine is a potent antiplatelet agent, which activates adenylate cyclase, inhibits platelet Ca(2+) mobilization, TXB(2) production as well as suppresses COX-1 enzyme activity. Sanguinarine may have therapeutic potential for treatment of cardiovascular diseases related to platelet aggregation.

Effects of root-end filling materials and eugenol on mitochondrial dehydrogenase activity and cytotoxicity to human periodontal ligament fibroblasts.

Various root-end filling materials have been used to prevent the entry of root-canal pathogens into periapical regions. Five root-end filling materials were compared regarding the cytotoxicity, apoptosis, and mitochondrial dehydrogenase (MDH) activities of human periodontal ligament (PDL) fibroblasts, with the use of a novel transwell culture system. Exposure to IRM (a ZnO eugenol-based intermediate restorative material), a 2-ethoxybenzoic acid cement (Super EBA), and amalgam for 3 days inhibited the MDH activity of PDL fibroblasts as indicated by decrease in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) reduction by 97%, 95%, and 51%, respectively. Evident suppression of MTT reduction by amalgam and glass ionomer cement (GIC) was noted after 5 days of exposure, with 73% and 46% of inhibition, respectively. Mineral trioxide aggregates (MTA) showed little effect on MDH activity. IRM and Super EBA were cytotoxic to PDL fibroblasts as indicated by a trypan blue dye exclusion technique. GIC and amalgam showed mild cytotoxicity. IRM, GIC, and amalgam further induced apoptosis of PDL cells, as revealed by the presence of sub-G0/G1 DNA content in flow cytometric histogram. Twenty-four-hour exposure to IRM and Super EBA elevated the MDH activities to 156% and 117%, correspondingly, of that of control. Eugenol, a phenolic ingredient in Super EBA and IRM, also increases MDH activity of PDL fibroblasts by 45% and 51%, at concentrations of 0.5 and 1 mM. However, at concentrations higher than 0.5 mM, eugenol decreased the number of viable PDL fibroblasts. These results suggest that MTA is a biocompatible root-end filling material, followed by self-curing Fuji II GIC and amalgam. IRM and Super EBA ingredients induced marked cytotoxicity and transiently stimulate MDH activities, which is possibly due to their content of eugenol and induction of cellular adaptive response.

The induction of prostaglandin E2 production, interleukin-6 production, cell cycle arrest, and cytotoxicity in primary oral keratinocytes and KB cancer cells by areca nut ingredients is differentially regulated by MEK/ERK activation.

There are about 200-600 million betel quid (BQ) chewers in the world. BQ chewing is one of the major risk factor of hepatocarcinoma, oropharyngeal, and esophagus cancers in Taiwan, India, and Southeast Asian countries. Thus, the precise molecular mechanisms deserve investigation. We used cultured primary keratinocytes and KB cells, RT-PCR, flow cytometry, Western blotting, and ELISA to evaluate whether alterations in early gene expression is crucial in the carcinogenic processes of BQ. We observed the induction of c-Fos mRNA expression in human gingival keratinocyte (GK) and KB carcinoma cells by areca nut (AN) extract and arecoline. A maximal increment in c-fos gene expression was shown at about 30 min after challenge. AN extract (100-800 microg/ml) and arecoline (0.1-0.8 mM) also stimulated ERK1/ERK2 phosphorylation with a maximal stimulation at 5-10 min of exposure. Pretreatment by U0126 (30 microM), a MEK inhibitor, markedly inhibited the c-Fos, cyclooxygenase-2 (COX-2), and IL-6 mRNA expression of the KB epithelial cells. In addition, U0126 and PD98059 (50 microM) also decreased AN extract- and arecoline-associated PGE2 and IL-6 production in GK and KB cells. However, U0126 by itself arrested the cells in G0/G1 phase, but was not able to prevent AN- and arecoline-induced cell death or apoptosis. In contrast, U0126 enhanced the AN-induced apoptosis of KB cells. AN ingredients thus play a significant role in the pathogenesis of oropharyngeal cancer by activation of MEK1/ERK/c-Fos pathway, which promotes keratinocyte inflammation, cell survival, and affects cell cycle progression.