Immunopharmacology and InflammationIn vitro and in vivo anti-inflammatory activities of columbin through the inhibition of cycloxygenase-2 and nitric oxide but not the suppression of NF-κB translocation
Introduction
Recent advances in fundamental biomedical research have established an essential role for inflammation in mediating some human afflictions. This list of inflammatory diseases would run to over 100 each of which causes the deterioration of tissue in one or more parts of the body (Kharitonov, 2004, Nathan and Ding, 2010). Therefore, the inflammatory response must be actively ended when no longer needed to prevent unnecessary harmful biological processes. Mechanisms which serve to terminate inflammation include various cellular and immunological responses and could be initiated using potential anti-inflammatory compounds, which work specifically by inhibiting inflammatory components or activating transcription factors (Tak and Firestein, 2001).
Current anti-inflammatory drugs can inhibit inflammation as curative agents. These conventional drugs have not been successful to cure chronic inflammatory disorders. Natural products play a significant role in human health in relation to the prevention and treatment of inflammatory conditions. Therefore, there is a need for new and safe anti-inflammatory agents and one of the on-going research candidates are plant constituents used in herbal and traditional medicine (Nam, 2006, Read, 1995). Tinospora bakis Miers (Menisoermaceae), locally known in Sudan as “Erg-elhagar”, is used traditionally to treat headache and rheumatism (Broun and Massy, 1929, El Ghazali et al., 2003). Recently, Zafinindra et al. (2003) indicated that the aqueous extract of T. bakis roots has shown in vivo antipyretic effects. Alkaloidal extract of this plant have revealed also in vitro antimalarial activity against Plasmodium falciparum chloroquine-resistant strain W2 (Ouattara et al., 2006). Recently our laboratory confirmed that the plant ethanolic crude extract has potent immunosuppressive and antioxidant activities.
Biologically guided phytochemical study in our laboratory led to the isolation of columbin (Fig. 1), a diterpenoid furanolactone, in a bulk amount for the first time from the root of T. bakis (Koko et al., 2008, Koko et al., 2009). This is thus the fourth report of its occurrence in this family. Originally isolated from colombo root (Jateorhiza palmata Miers, Memspermaceae), columbin has more recently been found in seeds of Sphenocentrum Jollyanum (Memspermaceae). It has, however, been found once in an unrelated family, in Melothria maderaspatana Cogn. (Cucurbitaceae), so that it cannot be regarded as being entirely specific to the Menispermaceae family. Nevertheless, the occurrence of several oxides of columbin in another member of the family, Fdxzurru chloroleucu, suggests that this type of diterpenoid bitter principle may be fairly widespread in this plant group (Cava and Soboczenski, 1956, Cava et al., 1959, Chen et al., 1973, Gilbert et al., 1967, Hanuman et al., 1986, Ramstad et al., 1975). The crude drugs Jateorhiza columba, T. capillipes, T. sagittata, Dzoscoreophyllum cummznsiz, Sphenocentrum Jollyanum and M. maderaspatana contain columbin, and were used for traditional healing of various diseases including inflammatory ones (Cava and Soboczenski, 1956, Cava et al., 1959, Chen et al., 1973, Gilbert et al., 1967, Hanuman et al., 1986, Ramstad et al., 1975). However there were few reports on the biological activities of this bitter substance. Therefore, the current study was designed to investigate the mechanistic anti-inflammatory action of columbin in vitro, in silico and in vivo. We have also tested toxicity of the compound on normal mice and human normal liver cells (WRL-68).
Section snippets
Cell lines and reagents
RAW264.7 cell line was obtained from American Type Culture Collection (ATCC), USA. Dulbecco's Modified Eagle Medium (DMEM) both with and without phenol red, phosphate buffered saline and Hanks' balanced salt solution (HBSS), 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT), phosphate buffered saline (PBS) and Griess reagent were from Invitrogen (Carlsbad, USA). Foetal bovine serum (FBS), LPS from E. coli serotype 0111:B4, Indomethacin, l-NAME [l-NG-nitroarginine methyl ester
Effect of columbin on NO2− and PGE2 production and RAW cells viability
The induction of RAW 264.7 cells into an inflammatory state by treatment with LPS/IFN-γ caused significant increase in NO and PGE2 as shown in Fig. 2. As shown in Fig. 2A, columbin also inhibited the production of PGE2 in a dose-dependent manner. The breakdown product of secreted NO namely NO2− was detected in media at a mean concentration of 39.60 ± 3.32 μM. Cells that were not induced released trace amounts of NO. Columbin showed a dose-related inhibition of NO production in which significant
Discussion
The current study was designed to investigate the in vitro, in vivo and in silico anti-inflammatory activities of columbin. Although this compound was isolated previously (Atta-urRahman and Ahmad, 1988), to the best of our knowledge this is the first time it's isolation from the species T. barkis is reported. Moreover, this is the first time the compound was shown to inhibit COX-2 and NO.
Nitric oxide plays an important role in various inflammatory conditions where it is produced by the
Acknowledgment
The authors would like to express their utmost gratitude and appreciation to University of Malaya (HIR grant F00002-21001: Cell Biology and Drug Mechanism) for providing grant to conduct this study.
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2019, PhytomedicineCitation Excerpt :In addition, prolonged stimulation by these cytokines induces overexpression of iNOS, which in turn leads to aberrant secretion of NO. This contributes to pathological damage, toxicity, and apoptosis (Dimmeler and Zeiher, 1997). Because of these interest, several research groups have focused on identifying phytochemicals that regulate NO production (Abdelwahab et al., 2012 and Tunon et al., 2009). Plants contain diverse bioactive constituents, such as flavonoids, with a wide range of biological activities, e.g., antioxidant, anti-inflammatory, anticarcinogenic, anti-molluscidal, antimicrobial, anti-nociceptive, anti-spasmodic, anti-hepatotoxic, anti-inflammatory, antidiabetic, and antihypertensive properties (Grotewold, 2006).
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2017, Journal of EthnopharmacologyCitation Excerpt :There were two major compounds isolated from T. bakis, namely palmitine and columbin. Beauquesne and Lucienne (1938) reported the isolation of palmitine, while Abdelwahab et al. (2012) reported the biological activities of columbin isolated from this species. Tinospora species are of highly importance to the traditional healers since many years.
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2015, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life SciencesCitation Excerpt :Pharmacological studies showed that this diterpenoid has multiple attractive biological activities, including anti-carcinogenesis [2,3], anti-inflammatory [2,3], and anti-hyperlipidemia (by reducing cholesterol uptake) [4]. In vivo chemoprevention studies indicated that columbin was actively against human colon cancer carcinogenesis [5,6]. In addition to be bioactive, columbin may also impact the metabolism of co-administered drugs, since it was shown to affect the sleeping time of anesthetized mice [7].