ABSTRACT
Seungma-galgeun-tang (SGT) has been used for treatment of chronic diseases in the folk medicine recipe. Since nitric oxide (NO) is one of the major inflammatory parameters, we first studied the effects of SGT on NO production in lipopolysaccharide (LPS)-stimulated BV-2 microglia. SGT inhibited the secretion of NO in BV-2 microglia, without affecting cell viability. The protein level of inducible nitric oxide synthase (iNOS) was decreased by SGT and SGT also inhibited production of PGE(2) and expression of Cox-2. Proinflammatory cytokines, such as TNF-alpha, IL-1beta and IL-12, were inhibited by SGT in a dose-dependent manner and SGT blocked the activation of NF-kappaB, which was considered to be a potential transcription factor for the expression of iNOS, COX-2 and proinflammatory cytokines. SGT also blocked the degradation of IkappaB and activation (decrease of cytosolic p65) of NF-kappaB, p65. These results suggest that SGT could exert its anti-inflammatory actions by suppressing the synthesis of NO through inhibition of NF-B activity.
Subject(s)
Drugs, Chinese Herbal/pharmacology , Inflammation/prevention & control , Microglia/metabolism , NF-kappa B/metabolism , Animals , Base Sequence , Blotting, Western , Cell Line , Cyclooxygenase 2/biosynthesis , Cyclooxygenase 2/drug effects , DNA Primers , Dinoprostone/antagonists & inhibitors , Down-Regulation/drug effects , Hydrolysis , Inflammation/metabolism , Inflammation Mediators/antagonists & inhibitors , Mice , Nitric Oxide/biosynthesis , Nitric Oxide Synthase Type II/biosynthesis , RNA, Messenger/geneticsABSTRACT
Chan Su is a traditional Chinese medicine prepared from the dried white secretion of the auricular and skin glands of toads, and has been used as an Oriental drug. However, little is known about the effect of Chan Su on the growth of human cancer cells. This study was undertaken to investigate the underlying mechanism of Chan Su-induced apoptosis in a human bladder carcinoma cell line, T24. The effects of this compound were also tested on cyclooxygenase (COX) activity. Treatment of T24 cells with Chan Su resulted in the inhibition of viability and induction of apoptosis in a concentration-dependent manner, which was proved by trypan blue counts, DAPI staining, agarose gel electrophoresis and flow cytometric analysis. Apoptosis of T24 cells by Chan Su was associated with a down-regulation of anti-apoptotic Bcl-2 and Bcl-X(S/L) expression and an up-regulation of pro-apoptotic Bax expression. Chan Su treatment induced the proteolytic activation of caspase-3 and caspase-9, and a concomitant degradation of poly(ADP-ribose)-polymerase and beta-catenin protein. Furthermore, Chan Su decreased the levels of COX-2 mRNA and protein expression without significant changes in the levels of COX-1, which was correlated with an inhibition in prostaglandin E(2) synthesis. Taken together, these findings partially provide novel insights into the possible molecular mechanisms of the anti-cancer activity of Chan Su.