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      Water extract of Galla Rhois with steaming process enhances apoptotic cell death in human colon cancer cells

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          Abstract

          Background

          Galla Rhois has been considered to have medicinal properties against diarrhea, excessive sweating, bleeding, and chronic cough in Asian countries. Gallotannins, which are Galla Rhois-derived tannins, have been reported to possess biological and pharmacological activities, especially anticancer activity. In this study, we evaluated the effect of steaming at a temperature over 120 °C on the chemical constituents and biological activities of the water extract of Galla Rhois (GRE).

          Methods

          GRE was steamed at a temperature over 120 °C (AGRE), and its specific constituents were analyzed; the results were validated using a high-performance liquid chromatography–diode array detector system. To evaluate the anticancer effect of GRE and AGRE, cell viability assay, cell cycle analysis, and Western blot analysis were performed in HCT116 human colon cancer cells.

          Results

          Steaming markedly increased the contents of gallic acid and ellagic acid in GRE, and GRE or AGRE treatment reduced the viability of HCT116 cells. Notably, the steaming process enhanced the growth inhibitory effect of GRE in cancer cells. AGRE induced apoptosis through the activation of caspase-3, caspase-8, and caspase-9. Additionally, AGRE regulated the activation of mitogen-activated protein kinases including extracellular signal-regulated kinase, p38, and c-Jun NH2-terminal kinase, whereas GRE did not. However, both GRE and AGRE inhibited the activation of AKT.

          Conclusion

          Compared with GRE, AGRE is more potent in its ability to induce apoptosis in HCT116 cells; therefore, we suggest that the steaming process may be useful as a feasible method for improving the anticancer effect of GRE.

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          Most cited references32

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          Morphologic and biochemical hallmarks of apoptosis.

          Apoptosis is characterised by a series of typical morphological features, such as shrinkage of the cell, fragmentation into membrane-bound apoptotic bodies and rapid phagocytosis by neighbouring cells. This paper reviews the current knowledge on the molecular mechanisms of apoptosis as they relate to the morphologic hallmarks and their implications for the detection of apoptosis in cardiac tissue. Activation of cysteine proteases called caspases plays a major role in the execution of apoptosis. These proteases selectively cleave vital cellular substrates, which results in apoptotic morphology and internucleosomal fragmentation of DNA by selectively activated DNases. In response to several pro-apoptotic signals, mitochondria release caspase activating factors, that initiate an escalating caspase cascade and commit the cell to die. Members of the Bcl-2 oncoprotein family control mitochondrial events and are able to prevent, or induce, both apoptotic and non-apoptotic types of cell death. This suggests that different types of cell death share common mechanisms in the early phases, whereas activation of caspases determines the phenotype of cell death. Detection of apoptotic cells in tissue samples currently relies on the TUNEL assay. TUNEL-positive cardiomyocytes show morphological features of apoptosis and the typical ladder pattern in DNA electrophoresis. Thus, provided that the staining protocol is carefully standardised, this quantitative methodology provides reproducible results of the occurrence of cardiomyocyte apoptosis in cardiac samples. Recently, potentially more specific assays based on analysis of DNA fragmentation or demonstration of caspase activation have been developed. Applicability of these assays to demonstrate cardiomyocyte apoptosis should be tested.
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            MAPK signal specificity: the right place at the right time.

            Although the mechanisms that lead to activation of the Ras, extracellular-signal-regulated kinase mitogen-activated protein kinase (Ras/ERK-MAPK) signaling pathway have been studied intensively, the fundamental principles that determine how activation of ERK signaling can result in distinct biological outcomes have only recently received attention. Factors such as cell-surface receptor density, expression of scaffolding proteins, the surrounding extracellular matrix, and the interplay between kinases and phosphatases modulate the strength and duration of ERK signaling. Furthermore, the spatial distribution and temporal qualities of ERK can markedly alter the qualitative and quantitative features of downstream signaling to immediate early genes (IEG) and the expression of IEG-encoded protein products. As a result, IEG products provide a molecular interpretation of ERK dynamics, enabling the cell to program an appropriate biological response.
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              Steaming of ginseng at high temperature enhances biological activity.

              The present study was performed to evaluate the effect of steaming ginseng at a temperature over 100 degrees C on its chemical constituents and biological activities. Raw ginseng was steamed at 100, 110, and 120 degrees C for 2 h using an autoclave. The ginseng steamed at 120 degrees C was more potent in its ability to induce endothelium-dependent relaxation. Steaming the raw ginseng at 120 degrees C also remarkably increased the radical-scavenging activity. Ginsenosides F(4), Rg(3), and Rg(5), which were not present in raw ginseng, were produced after steaming. Ginsenosides Rg(3) and Rg(5) were the most abundant ginsenosides in the ginseng steamed at 120 degrees C, accounting for 39% and 19% of all ginsenosides, respectively.
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                Author and article information

                Contributors
                Journal
                Integr Med Res
                Integr Med Res
                Integrative Medicine Research
                Elsevier
                2213-4220
                2213-4239
                11 October 2016
                December 2016
                11 October 2016
                : 5
                : 4
                : 284-292
                Affiliations
                [0005]Korean Medicine (KM) Application Center, Korea Institute of Oriental Medicine (KIOM), Daegu, Korea
                Author notes
                [* ] Corresponding authors. Korean Medicine (KM) Application Center, Korea Institute of Oriental Medicine (KIOM), 70 Cheomdan-ro, Daegu 701-300, Korea. wkcho@ 123456kiom.re.kr jyma@ 123456kiom.re.kr
                [☆]

                Both authors contributed equally to this work.

                Article
                S2213-4220(16)30099-3
                10.1016/j.imr.2016.10.001
                5390415
                8f4501e9-066c-4cb3-96e5-51fc9b3e88cf
                © 2016 Korea Institute of Oriental Medicine. Published by Elsevier.

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 23 August 2016
                : 28 September 2016
                : 3 October 2016
                Categories
                Original Article

                ellagic acid,galla rhois,gallic acid,human colon cancer cells,steaming process

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