Auranofin is a US Food and Drug Administration (FDA)-approved anti-arthritis medication that functions as a thioredoxin reductase inhibitor. Spermidine, a polyamine present in marine algae, can exert various physiological functions. Herein, we examined the synergistic anticancer activity of auranofin and spermidine in hepatocellular carcinoma (HCC). Combined treatment with auranofin and spermidine suppressed cell viability more efficiently than either treatment alone in HCC Hep3B cells. The isobologram plotted by calculating the half maximal inhibitory concentration (IC50) values of each drug indicated that the two drugs exhibited a synergistic effect. Based on the analysis of annexin V and cell cycle distribution, auranofin and spermidine markedly induced apoptosis in Hep3B cells. Moreover, auranofin and spermidine increased mitochondria-mediated apoptosis by promoting mitochondrial membrane potential (Δψm) loss. Auranofin and spermidine significantly increased reactive oxygen species (ROS) production in Hep3B cells, and the blocking ROS suppressed apoptosis induced by spermidine and auranofin. In addition, auranofin and spermidine reduced the expression of phosphorylated phosphatidylinositol-3 kinase (PI3K) and protein kinase B (Akt), and PI3K inhibitor accelerated auranofin- and spermidine-induced apoptosis. Using ROS scavenger and PI3K inhibitor, we revealed that ROS acts upstream of auranofin- and spermidine-induced apoptosis. Collectively, our study suggests that combination treatment with auranofin and spermidine could afford synergistic anticancer activity via ROS overproduction and reduced PI3K/Akt signaling pathway.
Objectives: This study was intended to clarify how Jakyakkamchobuja-tang (hereinafter referred to JKBT) affects mice of C57BL/10 whose osteoarthritis was induced by papain. Methods: Osteoarthritis was induced in mice by injecting papain in the knee joint. Mice were divided into 4 groups (n=6). The normal group were not treated at all whereas the control group (OAC-control) were induced for osteoarthritis by papain and oral medicated with 200 ul of physiological saline per day. The positive comparison group (OAC-$Joins^{(R)}$) were injected with papain and after 7 days, 100 mg/kg of $Joins^{(R)}$ were medicated with 200 ul of physiological saline mixed. The experimental group (OAC-JKBT) were injected with papain and after 7 days were medicated with 400 mg/kg of JKBT mixed with 200 ul of physiological saline. OAC-$Joins^{(R)}$ and OAC-JKBT were oral medicated for each substance for a total of 4 weeks, once per day. After experiments (from 1 week after injection of papain to 4 weeks elapsed), the function of liver and kidney, inflammation cytokine values within serum, degree of revelation for inflammation cytokine genes, immune cells within blood, metabolism of arachidonic acid and amount of cartilage were measured and histopathological variations for knee joint structures were observed. Results: Functions of liver and kidney were not affected. IL-$1{\beta}$ (interleukin-$1{\beta}$), MCP-1 (monocyte chemoattractant protein-1) and TNF-${\alpha}$ (tumor necrosis factor-${\alpha}$) were significantly reduced and IL-6 (interleukin-6) was also reduced but not significantly. After analyzing inflammation cytokine in joints with mRNA (messenger ribonucleic acid), revelation of IL-6, TNF-${\alpha}$, COX-2 (cyclooxygenase-2) and iNOS-II (inducible nitric oxide synthase-II) were all significantly reduced. Revelation of IL-$1{\beta}$ gene was also reduced but not significantly. Neutrophil for WBC (white blood cell) within serum was significantly reduced; monocyte was also reduced but not significantly. PGE2 (prostaglandin E2), TXB2 (thromboxane B2) were significantly reduced and LTB4 (leukotriene B4) was also reduced but not significantly. Destruction of cartilage on micro CT (computed tomography)-arthrography was reduced but had no significant differences. In terms of histopathology, infiltration of inflammation, proliferation of synovial membrane, subsidence of cartilage and bone due to penetration of excessive formation of synovial cell and destruction of cartilage were small (H&E (hematoxylin and eosin), safranine O staining). Conclusions: Based on these results, Jakyakkamchobuja-tang (JKBT) is believed to be useful for suppressing the progress of osteoarthritis and its treatments because of its anti-inflammatory effects and alleviation of pain with histopathological effective efficacy.
This study aimed to understand the physiological status of the beet armyworm, Spodoptera exigua at low developmental threshold temperature (LTT) through analysis of gene-expression patterns associated with different functions (metabolism, nervous system, immune, and stress). The estimated LTTs for egg, larval, and pupal developments varied with $5.5{\sim}11.6^{\circ}C$. Larvae were able to develop at the lower temperatures than eggs and pupae. However, the physiological LTT ($15^{\circ}C$) in the fifth instar was much higher than the estimated LTT ($10.3^{\circ}C$). Gene expression patterns estimated by a quantitative RT-PCR (qRT-PCR) indicate that most genes in different functional groups increased their expressions with increase of larval instars. In the same fifth instar, as the treatment temperatures increased, the gene expression levels increased. Moreover, the newly molted fifth instar larvae were different in their gene expression rates according to their previous culturing temperatures. Most gene expressions were suppressed in the fifth instar larvae at the physiological LTT ($15^{\circ}C$). However, the larvae at $15^{\circ}C$ gradually exhibited significant increase in the gene expression rates with rearing time just like those at the higher temperature. These results suggest that S. exigua at LTT exhibits a typical gene expression pattern with maintaining significantly suppressed levels.
Soyfoods have potential roles in the prevention and treatment of chronic diseases, most notably cancer, osteoporosis, and heart disease. There is evidence that carcinogenesis are supressed by isolated soybean derived products in vivo such as a protease inhibitor, phytic acid, saponins and isoflavones. It is believed that supplementation of human diets with soybean products markedly reduces human cancer mortality rates. Especially, recent papers recognize the potential benefit of soybean isoflavone components for reducing the risk of various cancers. Isoflavones exhibit a multitude of medicinal effects that influence cell growth and regulation, which may have potential value in the prevention and treatment of cancer. In addition to potential biological effects, soybean isoflavones have the important physiological functions such as the induction of Bradyrizobium japonicum nod genes and the responses of soybean tissues to infection by Phytophthora megasperma as well as biochemical activities such as antifungal and antibacterial actions. Genistin, daidzin, glycitin and their aglycone (genistein, daidzein, glycitein) are the principal isoflavones found in soybean. Malonyl and acetyl forms have also been detected but they are thermally unstable and are usually transformed during the processing in glucoside form. Most soy products, with the exception of soy sauce, alcohol-extracted soy protein concentrate, and soy protein isolate, have total isoflavone concentrations similar to those in the whole soybean. Soybean-containing diets inhibit mammary tumorigenesis in animal models of breast cancer, therefore, it is possible that dietary isoflavones are an important factor accounting for the lower incidence and mortality from breast cancer. Of the total soybean seed isoflavones, $80\~90\%$ were located in cotyledons, with the remainder in the hypocotyls. The hypocotyls had a higher concentrations of isoflavones on a weight basis compared with cotyledons. Isoflavone contents were influenced by genetics, crop years, and growth locations. The effect of crop year had a greater impact on the isoflavone contents than that of location. The climate condition might be the attribution factor to variation in isoflavone contents. Also, while the isoflavone content of cotyledons exhibited large variations in response to high temperature during seed development, hypocotyls showed high concentration in isoflavone content. So, it is concluded that one of the factors affecting isoflavone content in soybean seeds is temperature during seed development. High temperature, especially in maturity stage, causes lower isoflavone content in soybean seed. It is also suggested that there may exist a different mechanism to maintain isoflavone contents between cotyledon and seed hypocotyls. In a conclusion, soy foods may be able to have a significant beneficial impact on public health.
Proceedings of the Korea Environmental Mutagen Society Conference
/
2002.11a
/
pp.80-88
/
2002
Recent studies demonstrate that collagen IV selectively pro-motes the repair of physiological processes in sublethally injured renal proximal tubular ceils (RPTC). We sought to further define the mechanisms of cell repair by measuring the effects of toxicant injury and stimulation of repair by L-ascorbic acid-2-phosphate (AscP), exogenous collagen IV, or function-stimulating integrin antibodies on the expression and subcellular localization of collagen-binding integrins (CBI) in RPTC. Expression of CBI subunits ${\alpha}_1$, ${\alpha}_2$, and ${\beta}_1$ in RPTC was not altered on day 1 after sublethal injury by S-(1,2-dichlorovinyl)-L-cysteine (DCVC). On day 6, expression of ${\alpha}_1$ and ${\beta}_1$ subunits remained unchanged, whereas a 2.2-fold increase in ${\alpha}_2$ expression was evident in injured RPTC. CBI localization in control RPTC was limited exclusively to the basal membrane. On day 1 after injury, RPTC exhibited a marked inhibition of active $Na^+$ transport and a loss of cell polarity characterized by a decrease in basal CBI localization and the appearance of CBI on the apical membrane. On day 6 after injury, RPTC still exhibited marked inhibition of active $Na^+$ transport and localization of CBI to the apical membrane. However, DCVC-injured RPTC cultured in pharmacological concentrations of AscP (500 ${\mu}$M)or exogenous collagen IV (50 ${\mu}$g/ml) exhibited an increase inactive $Na^+$ transport, relocalization of CBI to the basal membrane, and the disappearance of CBI from the apical membrane on day 6. Function-stimulating antibodies to CBI ${\beta}_1$ did not promote basal relocalization of CBI despite stimulating the repair of $Na^+$/$K^+$-ATPase activity on day 6 after injury. These data demonstrate that DCVC disrupts integrin localization and that physiological repair stimulated by AscP or collagen IV is associated with the basal relocalization of CBI in DCVC-injured RPTC. These data also suggest that CBI-mediated repair of physiological functions may occur independently of integrin relocalization.
We conducted this study to investigate the morphological characteristics and physiological effects of mulberry leaves and fruits with wild varieties. Morphologically sampling materials were showed Morus bombycis type and Morus alba type characteristics. Water contents of mulberry fruits were higher than that of mulberry leaves. Sugar degree of sampling materials was 5.1∼22.7 Brix %. The yield of 85% MeOH extract from freezing dried Mulberry Fruits was 60.2% and Hexane, BuOH and H$_2$O fraction from MeOH extract were 2.8%, 4.6%, 92.3% respectively. In the cholesterol-induced hyperlipidemia rats administered with subfraction of mulberry fruit extract, total cholesterol and serum triglyceride were decreased in the MeOH extract group and H$_2$O soluble fraction group. Also H$_2$O soluble fraction group decreased GPT, GOT and LDH level. Therefore, the above results suggested that mulberry fruit with wild varieties can help to maintain normal liver functions and to protect hyperlipidemia.
Rice (Oryza sativa L.) is one of the major cereal crops for consumption by the world’s population. Recently, various colored rice, such as white, red, brown, green, and black rice, have caught the attention of world consumers. The commercial name ‘black rice’ contains a high amount of anthocyanins in pericarp, which increases nutritional value. Moreover, anthocyanin in black rice possesses biomedical properties, including anti-oxidant, anti-cancer, and anti-inflammatory effects in humans. In genetics, black rice has a dominant PURPLE PERICARP (Prp) trait governed by two genes, Pb and Pp, which are involved in the synthesis of cyanidin-3-O-glucoside (C3G). Since the publication of a report by Nagai at 1921, the genetics and physiological studies of black rice driven by Prp traits are still unable to understand the relevant genes and their roles. However, with the increased demand for anthocyanin-rich black rice as a functional food for human health, it has become urgent to develop highyielding anthocyanin-rich varieties of rice. We explored many years in the genetics of purple pericarp trait, anthocyanin biosynthesis in pericarp during seed development, and, consequently, their products in relation to different physiological and agronomic traits. In this review, we summarized the anthocyanin biosynthesis in pericarp, emphasizing the inheritance pattern of the trait and functions of their products on different physiological and agronomic traits, including the yield of black rice.
Hepatocyte growth factor (HGF), originally discovered and cloned as a powerful mitogen for hepatocytes, is a four kringle-containing growth factor which specifically binds to membrane-spanning tyrosine kinase, c-Met/HGF receptor. HGF has mitogenic, motogenic (enhancement of cell movement), morphogenic (e.g., induction of branching tubulogenesis), and anti-apoptotic activities for a wide variety of cells. During embryogenesis, HGF supports organogenesis and morphogenesis of various tissues, including liver, kidney, lung, gut, mammary gland, and tooth. In adult tissues HGF elicits an organotrophic function which supports regeneration of organs such as liver, kidney, lung, and vascular tissues. HGF is also a novel member of neurotrophic factor in nervous systems. Together with the preferential expression of HGF in mesenchymal or stromal cells, and c-Met/HGF receptor In epithelial or endothelial cells, the HGF-Met coupling seems to orchestrate dynamic morphogenic processes through epithelial-mesenchymal (or-stromal) interactions for organogenesis and organ regeneration. HGF or HGF gene may well become unique therapeutic tools for treatment of patients with various organ failure, through its actions to reconstruct organized tissue architectures. This review focuses on recently characterized biological and physiological functions integrated by HGF-Met coupling during organogenesis and organ regeneration.
Circadian rhythms can be seen in a variety of physiological functions in insects. Light is a powerful zeitgeber not only synchronizing but also modulating the rhythm to adjust insect's temporal structure to seasonal changes in the environmental cycle. There are two general effects of the length of light phase within 24 hr light cycles on the circadian rhythms, i.e., the modulation of free-running period and the waveform. Since the photoperiodic modulation of the free-running period is induced even in the clock mutant flies, per$\^$s/, the free-running period is not fully determined genetically. In crickets, the ratio of activity (a) and rest phase (p) under the constant darkness (DD) is clearly dependent on the photoperiod under which they have been kept. When experienced the longer photoperiod it becomes smaller. The magnitude of change in a/p-ratio is dependent on the number of cycles they experienced. The neuronal activity of the optic lobe in DD shows the a/p-ratio changing with the preceding photoperiod. These data suggest that a single circadian pacemaker stores and maintains the photoperiodic information and that there is a system that accumulates the effects of single photoperiod to cause greater effects.
Low-threshold T-type $Ca^{2+}$ channels are distinctive voltage-operated gates for external $Ca^{2+}$ entry around a resting membrane potential due to their low voltage activation. These phenomena have already been extensively studied due to their relevance in diverse physiological functions. Recently, three T-type $Ca^{2+}$ channel ${\alpha}$$_1$subunits were cloned and their biophysical properties were characterized after expression in mammalian expression systems. In this study, ${\alpha_IG} and {\alpha_IH}$ low-threshold $Ca^{2+}$ channels were expressed and characterized in Xenopus oocytes after adding 5' and 3'untranslated portions of a Xenopus ${\beta}$ globin to improve their expression levels. The added portions dramatically enhanced the expression levels of the ${\alpha_IG} and {\alpha_IH}$ T-type channels. When currents were recorded in 10 mM $Ba^{2+}$ as the charge carrier, the activation thresholds were about -60 mV, peak currents appeared at -20 mV, and the reversal potentials were between +40 and +45. The activation time constants were very similar to each other, while the inactivation time constants of the ${\alpha_IG}$ currents were smaller than those of ${\alpha_IH}$. Taken together, the electrophysiological properties of the ${\alpha_IG} and {\alpha_IH}$ channels expressed in Xenopus oocytes were similar to the previously reported characteristics of low-threshold $Ca^{2+}$ channel currents.
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