Anti-inflammatory Effects of Indomethacin's Methyl Ester Derivative and Induction of Apoptosis in HL-60 Cells

Abstract

Recent studies have reported that gastric, colon and other carcinogenic processes could be blocked by non-steroidal anti-inflammatory drugs (NSAIDs), through the inhibition of cyclooxygenase 2 (COX 2).1) Moreover, NSAIDs, such as celebrex, sulindac, indomethacin, and others, could induce apoptosis in tumor cells and act similarly to anticancer drugs. [1][2][3][4][5] Indomethacin is a well-known NSAID acting as a non-selective cyclooxygenase inhibitor, which is capable of inducing apoptosis in small-cell lung cancer and gastric cancer cells. [3][4][5] These facts imply that NSAIDs may be good lead compounds for inducing chemopreventive effects. In a previous study, the ester and g-lactone derivatives of indomethacin were found through a photo-irradiated process. 6) In this paper, the anti-inflammatory effects of indomethacin derivatives in LPS-stimulated RAW 264.7 cells and the cytotoxic effects in HL-60 cells were explored in efforts to discover new and potent chemopreventive agents.Nitric oxide (NO) induces mutagenesis and carcinogenesis through damaging DNA bases, and also has inflammatory effects such as enhancing COX activity, etc.7,8) Macrophages play major roles in inflammation, which can be activated by inducible nitric oxide synthase (iNOS) and COX 2 protein, and by the release of NO and prostaglandin E 2 (PGE 2 ) by lipopolysaccharide (LPS), an endotoxin. 9) Therefore, the NO and PGE 2 levels of LPS-stimulated murine macrophage RAW 264.7 cells are used as indicators of carcinogenesis and inflammation.The human promyelocytic leukemia cell line, HL-60, is usually used to detect the antitumor effects of new candidate compounds in in vitro cytotoxicity assays. Cell death can generally occur by either of two distinct mechanisms, necrosis or apoptosis. 10) Necrosis is the pathological process which occurs when cells are exposed to a serious physical or chemical insult. Apoptosis is the physiological process by which unwanted or useless cells are eliminated during development and other normal biological processes.10) Anticancer drugs can induce apoptosis in tumor cells, and this may decrease the side effects in patients. Consequently, compounds that can induce apoptosis in tumor cells with good potency are considered to be antitumor candidate drugs. In this paper, the cytotoxic effects of test samples were measured using the MTT assay, and the active components inducing apoptosis in leukemia cells were determined by biochemical analysis. MATERIALS AND METHODS Chemicals and CellsDimethyl sulfoxide (DMSO), MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide], LPS (Escherichia coli serotype 0127-8B), and other chemicals were purchased from Sigma Chemical (St. Louis, MO, U.S.A.). Dulbecco's modified Eagle medium (DMEM), RPMI 1640, fetal bovine serum (FBS), antibiotics, and L-glutamine were purchased from Gibco BRL (Grand Island, NY, U.S.A.). The murine macrophage cell line, RAW 264.7, and the promyelocytic leukemia cell line, HL-60, were obtained from American Type Cell Culture (ATCC; Rockville, MD,...

Marrubiin Inhibits Peritoneal Inflammatory Response Induced by Carrageenan Application in C57 Mice

“…Thus, a decrease in different blood cell types in the peritoneal fluid of treated animals, along with altered cell morphology, further supports the impact of M. vulgare extract on inflammatory processes (Figure 2D). Even more pronounced results were observed in peritoneal fluid cells obtained from animals treated with indometacin and carrageenan (Figure 2C), confirming the efficacy of this standard drug [13].…”

“…Indometacin and diclofenac, common positive control drugs, exert their anti-inflammatory effects by inhibiting prostaglandin synthesis, suppressing cyclooxygenase (COX), and modulating K + channel opening, which are crucial mediators of the inflammatory response. Moreover, these drugs often induce inflammatory cell apoptosis and inhibit neutrophil aggregation, partially mediated through prostaglandin signaling [13]. Consequently, a decrease in the number of inflammatory cells in the peritoneal cavity was observed in mice treated with carrageenan alone or in combination with diclofenac/indometacin (Figure 1).…”

“…MAR was isolated from Marrubium peregrinum L. (Lamiaceae) extract (hexane/Et2O/ethyl acetate/MeOH) via silica gel and Sephadex chromatography as a white powder. The structure was initially determined via NMR analysis (1D and 2D) and unequivocally confirmed through comparing 1 H and 13 C NMR chemical shifts (in CDCl3) with those reported in the literature [10]. Graphical 1 H and 13 C spectra are given in the Supplementary Materials (Figures S1-S3).…”

“…The structure was initially determined via NMR analysis (1D and 2D) and unequivocally confirmed through comparing 1 H and 13 C NMR chemical shifts (in CDCl3) with those reported in the literature [10]. Graphical 1 H and 13 C spectra are given in the Supplementary Materials (Figures S1-S3).…”

Identification of flurbiprofen and its photoproducts in methanol by gas chromatography–mass spectrometry

“…Recently in our laboratory, the methyl ester of indomethacin has been demonstrated to inhibit prostaglandin E 2 (PGE 2 ), NO production, inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX 2) protein expression. The ester also exhibited strong cytotoxicity in promyelocytic leukemia HL-60 cells (Chao et al, 2005). The pharmacological effects of FB and photoproducts are currently under investigation.…”