Gecko is a kind of traditional Chinese medicine with remarkable antineoplastic activity. However, undefined mechanisms and ambiguity regarding active ingredients limit new drug development from gecko. This study was conducted to assess anti-angiogenic properties of the aqueous extracts of fresh gecko (AG) or macromolecular components separated from AG (M-AG). An enzyme-linked immunosorbent assay (ELISA) approach was applied to detect the vascular endothelial growth factor (VEGF) secretion of the tumor cells treated with AG or M-AG. The effect of AG or M-AG on vascular endothelial cell proliferation and migratory ability was analyzed by tetrazolium dye colorimetric method, transwell and wound-healing assays. Chick embryo chorioallantoic membrane (CAM) assays were used to ensure the anti-angiogenic activity of M-AG in vivo. The results showed that AG or M-AG inhibited the VEGF secretion of tumor cells, the relative inhibition rates of AG and M-AG being 27.2% and 53.2% respectively at a concentration of $20{\mu}L/mL$. AG and M-AG inhibited the vascular endothelial (VE) cell proliferation with IC50 values of $11.5{\pm}0.5{\mu}L/mL$ and $12.9{\pm}0.4{\mu}L/mL$ respectively. The VE cell migration potential was inhibited significantly (p<0.01) by the AG (${\geq}24{\mu}L/mL$) or M-AG (${\geq}12\mu}L/mL$) treatment. In vivo, neovascularization of CAM treated with M-AG was inhibited significantly (p<0.05) at a concentration of ${\geq}0.4{\mu}L/mL$. This study provided evidence that anti-angiogenesis is one of the anti-tumor mechanisms of AG and M-AG, with the latter as a promising active component.
70% ethanolic extracts were prepared from the three mutant rice cultivars with giant embryo termed Shinsunchal-giant embryonic rice, Whachung-giant embryonic rice and Nampung-giant embryonic rice, and its antioxidative and antimutagenic properties were evaluated and compared. For analysing antioxidativity, various antioxidative indices, such as electron donating ability to DPPH radical, scavenging capacity to hydroxyl radicals generated by Fenton reaction, scavenging capacity to superoxide radicals generated by HPX/XOD system, inhibitory effect on autoxidation of linoleic acid and inhibitory effect on membrane lipid peroxidation derived from rabbit erythrocyte ghost, were determined. For analysing antimutagenicity, suppressive effects on mutagenesis induced by the chemical mutagen, mitomycin C, were measured using E. coli PQ 37 as a indicator cell. The results showed that for both antioxidativity and antimutagenicity the giant embryonic rices were more effective compared to the general cooking rice, Among the giant embryonic rice cultivars, Nampung-giant embryonic rice tended to be most effective, showing its scavenging activity to DPPH radical, superoxide radical and hydroxyl radical, and inhibitory activity to lipid peroxidation was 2,3-, 3,3-, 1.7-, and 2.5-fold greater than those of normal rice, respectively.
The human muscarinic acetylcholine receptor (mAChR) subtypes Hml, Hm2 and Hm3 have been expressed in insect cells (Spodoptera frugiperda, Sf9) using the baculovirus expression system. Expression of relevant DNA, transcript and receptor proteins was identified by PCR, Northern blotting and [$^{3}H$]QNB binding, respectively. As assessed by [$^{3}H$]QNB binding sites, yields of muscarinic receptors in membrane preparations in this study were as about 5-20 times high as those in mammalian cells reported in previous studies. The [$^{3}H$]QNB competition binding studies with well-known subtype-selective mAChR antagonists showed that the receptors expressed in Sf9 cells retain the pharmacological characteristics expected for the ml , m2 and m3 muscarinic receptors. The ml-selective antagonist, pirenzepine, displayed a considerably higher affinity for Hml by 110-fold and 35-fold than for Hm2 and Hm3, respectively, The m2-selective methoctramine displayed a significantly higher affinity for Hm2 than for Hml and Hm3 (10- and 26-fold, respectively). p-F-HHSiD exhibited high affinity for Hm3 that is not significantly different from those for Hml, but 66-fold higher than its affinity for Hm2. The functional coupling of the recombinant receptors to second messenger systems was also examined. While both Hml and Hm3 stimulated phosphoinositide hydrolysis upon activation by carba-chol, Hm2 produced no response. On the other hand, activation of mAChRs induced the inhibition of forsko-lin-stimulated cyclic AMP formation in Hm2-expressing cells, whereas the significant dose-dependent increase in or poor response on cyclic AMP formation were produced in Hml or Hm3-expressing cells, respectively. These results indicate the differential coupling of recombinant Hml, Hm2 and Hm3 receptors expressed in SF9 cells to intracellular signalling system.
$^{123}I$, which is applied for the thyroid and other in vivo kinetic study, has a special role in life sciences. The 159 KeV $\gamma-ray$ from $^{123}I$ is almost ideally appropriate for the current imaging instrumentation. Its decay mode (electron capture) and short half-life (13.3 hr) reduced the burden of radiation dose to the patients, and its chemical property makes it easy to synthesize the labelling compounds. In this experiment, the production of $^{123}I$ via the nuclear reaction $^{124}Te(p,2n)^{123}I$ with 28 MeV protons was sutdied. $TeO_2$ is used as a target material, because it has good physical properties. The target was prepared with $TeO_2$ powder and was molten into a ellipsoidal cavity (a=14 mm, b=10 mm, $270.8mg/cm^2$ thick) of pure platinum. The irradiation was carried out in the external proton beam with incident energies range from 28 MeV to 22 MeV, and current was $30{\mu}A$. The loss of $TeO_2$ target was significantly reduced by using $4\pi-cooling$ system in irradiation. The dry distillation method was adopted for the separation of $^{123}I$ from irradiated target, and when it was kept 5 minutes at $780^{\circ}C$, its result was quantitative. The loss of the target material $(TeO_2)$ was below 0.2% for each production run and $^{123}I$ from the dry distillation apparatus was captured with 0.01 N NaOH in $Na^{123}I$ form, then the pH of the solution was adjusted to $7.5\sim9.0$ with HC1/NaOH. The $Na^{123}I$ solution was passed through $0.2{\mu}m$ membrane filter, and sterilized under high pressure and temperature for 30 minutes. The production of $^{123}I$ is acceptable for clinical application based on the quality of USP XXI.
Copolyimide membranes of different chemical structure based on pyromellitic dianhydride (PMDA)/methylendianiline(MDA) were prepared by varying their chemical compositions with adding meta-phenylendiamine (MPD), para- phenylendiamine (PPD), 2,4,6- trimethyl-1,3-phenylenediamine(TriMeMPD) as a co-monomer. The $N_2$ and $O_2$ permeation properties are qualitatively correlated to specific free volume and intersegmental distance of membrane. The partial replacement of MDA with MPD or PPD caused in the PMDA/MDA based membranes increase in density, and decrease in free volume, d-spacing, consequently resulted in decreased permeability coefficient. In the case of TriMeMPD, opposite results were observed. In all membranes, the permeability coefficients were pressure independent, and membranes which have high permeability coefficient showed low $N_2/O_2$ ideal separation factor as an usual manner. The permeability coefficient also increased with temperature and $N_2/O_2$ ideal separation factor decreased. The increase ratio of the $N_2$ permeability coefficient was larger than that of $O_2$.
A membrane-like $H_3PMo_{12}O_{40}$-polysulfone film was prepared by blending $H_3PMo_{12}O_{40}$ with polysulfone using dimethylformamide as a common solvent. SEM and EDX analysis showed that $H_3PMo_{12}O_{40}$ was uniformly and finely distributed in the film catalyst. The ESCA measurement also revealed that the oxidation state of Mo was not changed. The $H_3PMo_{12}O_{40}$-polysulfone catalyst showed lower activity for acid-catalyzed reaction and higher activity for oxidation reaction than $H_3PMo_{12}O_{40}$ in ethanol conversion reaction. The oxidation activity of the film catalyst was about 10 times higher than $H_3PMo_{12}O_{40}$. The decrease of acidic activity was due to DMF strongly adsorbed in acid sites of $H_3PMo_{12}O_{40}$, whereas the increase of oxidation activity was mainly due to uniform distribution of $H_3PMo_{12}O_{40}$. Adsorption results showed that the surface character of $H_3PMo_{12}O_{40}$ was drastically increased, while the bulk property of that was almost same after blending. It is suggested that the control of surface/bulk property as well as acid/redox property of heteropoly acid would be possible by blending it with a polymer.
Sanguinarine is a benzophenanthridine alkaloid originally isolated from the roots of Sanguinaria canadensis. It has multiple biological activities (e.g., antioxidant and antiproliferative) and immune-enhancing potential. In this study, we explored the proapoptotic properties and modes of action of sanguinarine in human hepatocellular carcinoma HepG2 cells. Our results revealed that sanguinarine inhibited HepG2 cell growth and induced apoptosis in a dose-dependent manner. The induction of apoptosis by sanguinarine was associated with the up-regulation of Fas and Bax, the release of cytochrome c from the mitochondria to the cytosol, and the loss of the mitochondrial membrane potential. In addition, sanguinarine activated caspase-9 and -8, initiator caspases of the intrinsic and death extrinsic pathways, respectively, and caspase-3, accompanied by proteolytic degradation of poly (ADP-ribose) polymerase. Sanguinarine also triggered the generation of reactive oxygen species (ROS). The elimination of ROS by N-acetylcysteine reversed sanguinarine-induced apoptosis. Furthermore, sanguinarine induced the dephosphorylation of Akt and the phosphorylation of mitogen-activated protein kinases, including extracellular signal-regulated kinase (ERK), c-jun N-terminal kinase (JNK), and p38. The growth inhibition was enhanced by the combined treatment of sanguinarine with a phosphatidylinositol 3'-kinase (PI3K) inhibitor and an ERK inhibitor but not JNK and p38 inhibitors. Overall, our data indicate that the proapoptotic effects of sanguinarine in HepG2 cells depend on ROS production and the activation of both intrinsic and extrinsic signaling pathways, which is mediated by blocking PI3K/Akt and activating the ERK pathway. Thus, our data suggest that sanguinarine may be a natural compound with potential for use as an antitumor agent in liver cancer.
The contents of bioactive materials (e.g. polyphenolics compounds, flavonoids, minerals, and fatty acids) and antioxidative activities (DPPH (${\alpha}$,${\alpha}$'-diphenyl-${\beta}$-picrylhydrazyl) free radical scavenging activity, peroxidation of linoleic acid and rat hepatocyte microsome, and Fe/Cu reducing power) were tested by in vitro experimental models using water, ethanol and methanol extracts of leaves (TOL) and fruits (TOF) from Thuja orientalis. Methanol extract from TOL showed the highest extraction yield (12.90%) as well as contents of polyphenolic compounds (16.02%) and flavonoids (0.25%). Major minerals were Ca, K, and Mg. Major fatty acids were palmitic and lauric acids in TOL and palmitic and decanoic acids in TOF. In oxidation of in vitro models using DPPH free radical scavenging activity, Fe/Cu reducing power, $Fe^{2+}$/ascorbate-induced linolenic acid peroxidation by ferric thiocyanate and thiobarbituric acid (TBA) methods, and autooxidation of rat hepatic microsomes membrane, anti-oxidative activities were stronger in all extracts of TOL than in those of TOF in a dose-dependent manner. From these results, methanol extract of TOL was shown to have the most potent anti-oxidative properties and the highes content of antioxidative compounds such as polyphenolic compounds and flavonoids.
Kim, Su-Gwan;Kim, Hyun-Ho;Kim, Chang-Hyun;Kim, Do-Kyung
Journal of the Korean Association of Oral and Maxillofacial Surgeons
/
v.32
no.3
/
pp.200-208
/
2006
Amino acids are required for protein synthesis and energy sources in all living cells. The amino acid transport system L is a major nutrient transport system that is responsible for $Na^+$-independent transport of neutral amino acids including several essential amino acids. In malignant tumors, the L-type amino acid transporter 1 (LAT1), the first isoform of system L, is highly expressed to support tumor cell growth. In the present study, the expression and functional characterization of amino acid transport system L were, therefore, investigated in Saos2 human osteogenic sarcoma cells. RT-PCR and western blot analyses have revealed that the Saos2 cells expressed the LAT1 and the L-type amino acid transporter 2 (LAT2), the second isoform of system L, together with their associating protein heavy chain of 4F2 antigen (4F2hc) in the plasma membrane, but the expression of LAT2 was very weak. The uptakes of [${14}^C$]L-leucine by Saos2 cells were $Na^+$-independent and were completely inhibited by the system L selective inhibitor, 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH). The affinity of [${14}^C$]L-leucine uptake and the inhibition profiles of [${14}^C$]L-leucine uptake by various amino acids in the Saos2 cells were comparable with those for the LAT1 expressed in Xenopus oocytes. The majority of [${14}^C$]L-leucine uptake is, therefore, mediated by LAT1 in the Saos2 cells. These results suggest that the transports of neutral amino acids including several essential amino acids into Saos2 human osteogenic sarcoma cells are for the most part mediated by LAT1. Therefore, the Saos2 human osteogenic sarcoma cells are excellent tools for examine the properties of LAT1. Moreover, the specific inhibition of LAT1 in tumor cells might be a new rationale for anti-tumor therapy.
Cajanus indicus is a herb with medicinal properties and is traditionally used to treat various forms of liver disorders. Present study aimed to evaluate the effect of a 43 kD protein isolated from the leaves of this herb against chloroform induced hepatotoxicity. Male albino mice were intraperitoneally treated with 2mg/kg body weight of the protein for 5 days followed by oral application of chloroform (0.75ml/kg body weight) for 2 days. Different biochemical parameters related to physiology and pathophysiology of liver, such as, serum glutamate pyruvate transaminase and alkaline phosphatase were determined in the murine sera under various experimental conditions. Direct antioxidant role of the protein was also determined from its reaction with Diphenyl picryl hydraxyl radical, superoxide radical and hydrogen peroxide. To find out the mode of action of this protein against chloroform induced liver damage, levels of antioxidant enzymes catalase, superoxide dismutase and glutathione-S-transferase were measured from liver homogenates. Peroxidation of membrane lipids both in vivo and in vitro were also measured as malonaldialdehyde. Finally, histopathological analyses were done from liver sections of control, toxin treated and protein pre- and post-treated (along with the toxin) mice. Levels of serum glutamate pyruvate transaminase and alkaline phosphatase, which showed an elevation in chloroform induced hepatic damage, were brought down near to the normal levels with the protein pretreatment. On the contrary, the levels of anti-oxidant enzymes such as catalase, superoxide dismutase and glutathione-S-transferase that had gone down in mice orally fed with chloroform were significantly elevated in protein pretreated ones. Besides, chloroform induced lipid peroxidation was effectively reduced by protein treatment both in vivo and in vitro. In cell free system the protein effectively quenched diphenyl picryl hydrazyl radical and superoxide radical, though it could not catalyse the breakdown of hydrogen peroxide. Post treatment with the protein for 3 days after 2 days of chloroform administration showed similar results. Histopathological studies indicated that chloroform induced extensive tissue damage was less severe in the mice livers treated with the 43 kD protein prior and post to the toxin administration. Results from all these data suggest that the protein possesses both preventive and curative role against chloroform induced hepatotoxicity and probably acts by an anti-oxidative defense mechanism.
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