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PubMed2002

In vitro cytotoxicity of the organophosphorus insecticide methylparathion to FG-9307, the gill cell line of flounder (Paralichthys olivaceus).

FG-9307, a cell line derived from the gill of flounder Paralichthys olivaceus, was used to determine the acute cytotoxic effects of the organophosphorus insecticide methylparathion. The cytotoxic effects of methylparathion were initially measured by three endpoint systems: neutral red (NR) uptake assay, tetrazolium (MTT) assay, and cell protein assay. Results indicated that concentrations of methylparathion ranging from 5 microg/ml to 60 microg/ml were toxic, and there was no significant difference in cytotoxic effects between the three test systems. Thus, the FG-9307 cell line is one of several choices for evaluating the acute toxicities of organophosphorus insecticides such as methylparathion. The ultrastructure of the cells was also studied. It was found that the ultrastructure of the cells was markedly altered by methylparathion, as evidenced by dilation of mitochondria, breakdown of rough endoplasmic reticulum, nuclear necrosis, and production of numerous lysosomes and lipid vacuoles. This appears to be the first report that a marine fish cell line can be used for acute in vitro cytotoxicity evaluation of methylparathion.

Animals↗
PubMed2002

In vitro lead-induced cell toxicity and cytoprotective activity of fetal calf serum in human fibroblasts.

The underlying mechanisms by which lead ions produce their deleterious effects prior to the onset of clinical symptoms are incompletely understood. This study aimed to assess lead-induced cell toxicity mechanisms by focusing on the effects of the metal on cell growth, DNA synthesis, cellular ATP, intracellular hexosaminidase activity and lysosomal function, and examine the possible cytoprotective role of fetal calf serum (FCS). Several human dermal cultured fibroblast lines were exposed to Pb (400 microM) for 1-6 days with 2, 5, and 10% FCS. The earliest toxic effect of Pb was significant inhibition of DNA synthesis after 24 h direct exposure; this harmful effect was not progressive during the first 3 days, but worsened clearly on the 4th day regardless of the FCS concentration. Atime-dependent depletion of intracellularATP content was also caused by ionic lead, thereby compromising the cell energy charge which precedes cell death. Fibroblast growth was progressively and significantly inhibited from day 2 onwards; the greatest noxious effect was observed in the presence of 2% FCS: 49% reduction in cell proliferation after 5 days. Lead salts produced loss of cell adhesion to the culture dish which worsened from the 2nd day and was more pronounced when FCS in growth medium was decreased. Toxic actions on lysosomal membrane integrity provoked a decrease in neutral red uptake (NRU) which was exposure time-dependent and more marked with 2% FCS. In contrast, increased relative NRU (to 20% at 4 days), suggestive of endocytosis-induced lysosome enlargement, was observed in Pb-exposed cells. Intracellular hexosaminidase activity was not negatively affected until 5 days after exposure to Pb salts. FCS had a significant cytoprotective effect on Pb-induced toxicity.

Adenosine Triphosphate↗
PubMed1997

A possibility for new evaluating method of cytotoxicity by using heat shock protein assay.

To determine whether heat shock proteins can be utilized as a biomarker for cytotoxicity of dental materials the induction of synthesis of heat shock proteins by mercuric chloride was examined. To analyse the synthesis of heat shock proteins, HeLa cells were labelled with [35S] methionine, and the labelled proteins were separated by SDS-PAGE and autoradiographed. Incubation of the cells in a medium containing 1.25 to 40 microM mercuric chloride markedly increased the synthesis of HSP70. At 20 or 40 microM mercuric chloride in medium, HeLa cells synthesized HSP70 at 2 h after exposure, maximally at 4-7 h, and gradually diminished thereafter. Examination of the cytotoxicity of mercuric chloride by the conventional neutral red uptake assay revealed a reduction of uptake of the dye in the presence of mercuric chloride at 40 microM and above. These findings suggest that the induction of synthesis of HSP70 is one of the most sensitive cellular responses caused by mercury ion, and the heat shock protein assay can be utilized for evaluation of the cytotoxicity of dental materials.

Journal Article↗
PubMed2002

Effects of heterocyclic and tertiary permeant amines on the electron transfer in thylakoid membranes.

The effect of low concentrations (up to 50 muM) of lipophilic permeant amines on the electron transfer in thylakoid membranes of pea chloroplasts has been investigated. In the presence of heterocyclic amines (9-aminoacridine and neutral red), the electron transfer, initiated from H(2)O to PS I acceptors, has been shown to be inhibited to a level amounting to less than 50% of control, this taking place for both the basal (at alkaline pH) and the gramicidin-uncoupled transport (at pH 6.5-8.5). Under the same conditions, tertiary amines (dibucaine, tetracaine) cause only a 10-15% inhibition of transport. With all the amines, the rate of electron transport from H(2)O to DCBQ, PS II acceptor is decreased to 80-90% of control at pH above 8.0, but this effect is completely removed when pH is lowered to 7.7-6.5. In the region of PS I, all the amines accelerate the basal transport, but do not influence the uncoupled electron transfer. A conclusion has been drawn that, parallel with uncoupling, heterocyclic and tertiary amines also cause an inhibition of PS II, appearing at alkaline pH values. Additionally, heterocyclic amines seem to brake electron flow at the level of plastoquinone reduction.

Journal Article↗
PubMed2003

Indirect cytotoxicity evaluation of pseudowollastonite.

This study aimed to evaluate the cytotoxicity of substances leached by pseudowollastonite (CaSiO(3)). It has been previously shown that calcium (Ca(2+)) and silicate (SiO(3)(-)) ions are released from pseudowollastonite into biological solutions. Both of these ions are known to influence the biological metabolism of osteoblastic cells essential in the mineralization process and bone-bonding mechanism. The indirect toxicity evaluation was performed by extraction method, according to International Standard Organization (ISO). Pseudowollastonite pellets obtained by solid-state reaction were incubated, in culture medium, during 24, 48, 72 or 168 h at different concentrations (5, 10, 15, 50, 100, 200 mg/ml). The cytotoxicity of each extract in presence of human osteoblastic cell line (SaOS-2) was quantitatively assessed by measuring the viability (succinate dehydrogenase activity, MTT), the membrane integrity (the uptake of the neutral red by viable cells, NR) as well as the cell necrosis by measuring the lactate dehydrogenase (LDH) released in the culture medium. No significant alteration of membrane integrity or cell suffering was detectable. However, increased cell metabolism was observed for cells exposed to pseudowollastonite extract with longest extraction time (168 h). In conclusion, mineral elements leached by pseudowollastonite do not significantly affect the metabolism of osteoblastic cells.

Journal Article↗
PubMed2003

Testing the cytotoxicity of metal alloys used as magnetic prosthetic devices.

Technical magnetic materials are increasingly used for the development of magnetic retained dental prosthetic and orofacial epithetic devices. Since most of the magnets based on rare earth metals, such as samarium-cobalt based alloys have a high tendency for corrosion they were first coated by tin and then encapsulated by titanium. However, the high mechanical load particularly on dental devices may cause a rupture of the titanium capsule and the alloys contact directly biological fluids. Hence, it is important to know the cytotoxicity of these magnets to assess their potential effects on the surrounding tissue. In this study, the cytotoxicity of neodymium-iron-boron and samarium-cobalt (plain, tin and titanium coated) magnets was tested. First, magnets were incubated up to 7 days in culture medium to prepare extracts for cytotoxicity measurements. Changes in the surface morphology due to corrosion were visualized by scanning electron microscopy and analysis of the elemental composition. 3T3 mouse fibroblasts were cultured in the presence of extracts and their viability measured by neutral red and metabolic assays. To learn more about a possible toxic activity of the main components of magnets, salt solutions of different concentrations resembling those elements, which are main constituents of the magnets, were used. 3T3 fibroblasts were also cultured in direct contact with the materials and material induced effects on cell morphology and growth monitored by microscopy. As a result of this study it was found that samarium-cobalt magnets have a strong tendency for corrosion and exert a considerable cytotoxicity. Neodymium-iron-boron magnets have a lesser tendency for corrosion and are only moderate cytotoxic. Coating of samarium-cobalt magnets with tin or titanium makes the materials non-toxic. Application of salt solutions shows that cobalt has a tendency to be cytotoxic at higher concentrations, but enhances cell metabolism and proliferation at lower concentrations while the other magnet constituents had a lower or negligible cytotoxic potential.

Journal Article↗
PubMed2000

Successful separation of apoptosis and necrosis pathways in HaCaT keratinocyte cells induced by UVB irradiation.

UVB irradiation can induce apoptotic, necrotic, and differentiation pathways in normal human keratinocytes. The present study was undertaken to determine at what dose of UVB each of these pathways is induced and whether these pathways are distinct or overlapping. We have observed that UVB induces fragmentation of DNA in human HaCaT keratinocytes, in a bimodal manner. Low doses of UVB, 5-20 mJ/cm2, increase the levels of apoptosis as shown by increased levels of fragmented DNA, Fas, PARP, and FasL protein, and the number of apoptotic cells as assessed by FACS analysis. At higher doses of UVB (20 and 30 mJ/cm2) the number of apoptotic cells becomes reduced, as does the amount of Fas, PARP, and FasL protein. At these higher doses, cell viability is decreased as measured by DNA synthesis (BrdU labeling) neutral red uptake, which represents an increasing necrotic phenotype. Expression of markers of keratinocyte differentiation, involucrin, keratin K1, and keratin K10, are also observed to decrease with increasing UVB dose. These changes are accompanied by a further increase in DNA fragmentation. We conclude that low doses of UVB (5-20 mJ/cm2) induced an apoptotic pathway, whereas increasing doses (greater than 20 mJ/cm2) of UVB produce a direct necrotic effect and inhibit terminal differentiation.

Apoptosis↗
PubMed1997

Interactions of triethyltin-chloride (TET) with the energy metabolism of cultured rat brain astrocytes: studies by multinuclear magnetic resonance spectroscopy.

The effect of triethyltin-chloride (TET), a highly neurotoxic compound, on the cellular metabolism of rat brain astrocytes in vitro was examined by nuclear magnetic resonance (NMR) spectroscopy. 5-week-old cultures were exposed to TET (0.2-40 microM) either for (1) acute (3h), (2) 24 h, or (3) chronic treatment (8 d). Cells were labeled with 1-(13)C-glucose, cell extracts were prepared and 31P, 1H, and 13C spectra were analyzed. Cytotoxic effects of TET were assessed by vital dye uptake assay using neutral red (NR) and by exclusion of trypan blue (TB). Cells were examined ultrastructurally by electron microscopy. The data show that the major target of TET at concentrations already causing morphological effects on cultured astrocytes is not the energy metabolism, but that TET rather alters the intracellular concentrations of organic osmolytes, such as myo-inositol, taurine and hypotaurine, which are part of the control of ion and volume regulation and osmotic balance in astrocytes.

Animals↗
PubMed1997

Limitations of alginate gels as a culture model for the study of the effects of UVA radiation on human dermal fibroblasts.

Several studies were undertaken to develop three-dimensional (3-D) cell culture models that allow conditions closer to the in vivo situation. To this end, alginate gels were tested as a 3-D cell culture model that might be useful in the study of the effects of UVA on human dermal fibroblasts. Cell culture in alginate gels and the irradiation conditions were optimized. Results showed that optimized cultures in alginate gels experienced considerable cell death on UVA irradiation compared to the classical monolayer cell culture. Viability tests (cell counting and neutral red assay) were performed to show that only UVA-irradiated alginate gels were responsible for this cytotoxicity. The implication of oxygen species in the phototoxicity induced by ultraviolet light has already been described; for this reason we investigated whether oxygen species were involved in the cytotoxicity induced by alginate upon UVA irradiation. It appeared that superoxide anion is not implicated.

Alginates↗
PubMed2004

Uptake and cytotoxicity of chitosan molecules and nanoparticles: effects of molecular weight and degree of deacetylation.

PURPOSE: To evaluate the effects of molecular weight (Mw) and degree of deacetylation (DD) on the cellular uptake and in vitro cytotoxicity of chitosan molecules and nanoparticles. METHODS: Chemical depolymerization and reacetylation produced chitosans of Mw 213,000 to 10,000 and DD 88-46%, respectively. Chitosan was labeled with FITC and transformed into nanoparticles by ionotropic gelation. Uptake of chitosan by confluent A549 cells was quantified by fluorometry, and in vitro cytotoxicity was evaluated by the MTT and neutral red uptakeassays. RESULTS: Nanoparticle uptake was a saturable event for all chitosan samples, with the binding affinity and uptake capacity decreasing with decreasing polymer Mw and DD. Uptake fell by 26% when Mw was decreased from 213,000 to 10,000, and by 41% when DD was lowered from 88% to 46%; the uptake data correlated with the zeta potential of the nanoparticles. Uptake of chitosan molecules did not exhibit saturation kinetics and was less dependent on Mw and DD. Postuptake quenching with trypan blue indicated that the cell-associated chitosan nanoparticles were internalized, but not the cell-associated chitosan molecules. Chitosan molecules and nanoparticles exhibited comparable cytotoxicity, yielding similar IC50 and IC20 values when evaluated against the A549 cells. Cytotoxicity of both chitosan entities was attenuated by decreasing polymer DD but was less affected by a lowering in Mw. CONCLUSIONS: Transforming chitosan into nanoparticles modified the mechanism of cellular uptake but did not change the cytotoxicity of the polymer toward A549 cells. Chitosan DD had a greater influence than Mw on the uptake and cytotoxicity of chitosan nanoparticles because of its effect on the zeta potential of the nanoparticles.

Acetylation↗
PubMed2004

Assessment of the potential skin irritation of lysine-derivative anionic surfactants using mouse fibroblasts and human keratinocytes as an alternative to animal testing.

PURPOSE: The aim of this study was to identify new surfactants with low skin irritant properties for use in pharmaceutical and cosmetic formulations, employing cell culture as an alternative method to in vivo testing. In addition, we sought to establish whether potential cytotoxic properties were related to the size of the counterions bound to the surfactants. METHODS: Cytotoxicity was assessed in the mouse fibroblast cell line 3T6 and the human keratinocyte cell line NCTC 2544 using the MTT assay and uptake of the vital dye neutral red 24 h after dosing (NRU). RESULTS: Lysine-derivative surfactants showed higher IC50s than did commercial anionic irritant compounds such as sodium dodecyl sulfate, proving to be no more harmful than amphoteric betaines. The aggressiveness of the surfactants depended on the size of their constituent counterions: surfactants associated with lighter counterions showed a proportionally higher aggressivity than those with heavier ones. CONCLUSIONS: Synthetic lysine-derivative anionic surfactants are less irritant than commercial surfactants such as sodium dodecyl sulfate and hexadecyltrimethylammonium bromide and are similar to betaines. These surfactants may offer promising applications in pharmaceutical and cosmetic preparations, representing a potential alternative to commercial anionic surfactants as a result of their low irritancy potential.

Animal Testing Alternatives↗
PubMed2003

The effect of age of refrigerated conditioned medium on cell survivability in vitro.

Samples of a conditioned medium (CM), obtained from human ginvigal fibroblasts culture, were kept at 4 degrees C each for 2 weeks, 1, 2, 3 and 6 months. Their effectiveness in maintaining cell survival for their contact with L929 cells in 96 well tissue culture plates for 6 and 10 h at 4 degrees C was compared with Hank's Balanced Salt Solution (HBSS). Surviving cells were stained with neutral red dye and the dye intensity was measured spectrophotometrically at 540 nm. The target cells responded to all the above mentioned CM and the HBSS in the same manner at 6-h contact time. However, by 10-h contact time, the percentage of the target cells' survival was higher in the 2-week, 1- and 2-month CM than in HBSS, 3- and 6-month CM.

Animals↗
PubMed1995

Local cerebral glucose utilization and cytoskeletal proteolysis as indices of evolving focal ischemic injury in core and penumbra.

To ascertain the tempo of progression to irreversible injury in focal ischemia, we subjected halothane-anesthetized Sprague-Dawley rats to photochemically induced distal middle cerebral artery occlusion (dMCAO) combined with permanent ipsilateral and 1 h contralateral common carotid artery occlusions. Head temperature was maintained at 36 degrees C. At times centered at either 1.5 or 3 h post-dMCAO, the rate of local glucose metabolism (lCMRgl) was measured by 2-deoxyglucose autoradiography, and cytoskeletal proteolysis was assessed regionally by an immunoblotting procedure to detect spectrin breakdown products. At 1.5 h (n = 5), the cortical ischemic core was already severely hypometabolic (lCMRgl 15.5 +/- 10.8 mumol 100 g-1 min-1, mean +/- SD), whereas the cortical penumbral zone was hypermetabolic (69.0 +/- 9.7). (The lumped constant was verified to be unchanged by methylglucose studies). Neutral red pH studies at this time point showed that both the core and penumbral zones were equally acidotic. By 3 h post-dMCAO (n = 6), lCMRgl in the penumbral zone had fallen to low levels (15.4 +/- 2.2 mumol 100 g-1 min-1) equal to those of the ischemic core (16.7 +/- 4.5). Correspondingly, spectrin breakdown in the ischemic core was advanced at both 2 and 3.5 h post-dMCAO (36 +/- 18% and 33 +/- 18% of total spectrin, respectively), whereas in the penumbral zone spectrin breakdown was less extensive and more highly variable at both times (22 +/- 23% and 29 +/- 16%). We conclude that irreversible deterioration of the ischemic core, as evidenced by the onset of local cytoskeletal proteolysis, begins within 2 h of middle cerebral artery occlusion. In the ischemic penumbra, the transition from glucose hyper- to hypometabolism occurs by 3.5 h and is associated with a milder and more variable degree of spectrin breakdown.

Animals↗
PubMed1995

Critical subcellular targets of cisplatin and related platinum analogs in rat renal proximal tubule cells.

The clinical usefulness of chemotherapeutic agents containing the platinum moiety is often limited by their nephrotoxicity. To investigate the mechanism of nephrotoxicity, and to assess the effects of platinum analogs on specific organelles and basal protein synthesis, biochemical and ultrastructural analyses were performed in rat renal proximal tubule cells (RPTCs). Neutral red (NR) uptake was used to measure lysosomal function, and conversion of MTT to formazan used to assess mitochondrial function. Despite their differential toxicity, cisplatin, carboplatin and CI-973 caused similar progressive inhibition of specific functions, suggesting they may share a common mechanism of nephrotoxicity. Protein synthesis was the earliest indicator of toxicity, followed by NR uptake and MTT conversion. Fluorescent probes for lysosomes (acridine orange) and mitochondria (rhodamine 123) confirmed that cisplatin's toxicity to RPTCs was delayed and cumulative. Condensation of nucleolar components and fragmentation of RER were observed in RPTCs treated for as little as two hours. Since the nucleolus is the site of ribosome biogenesis, the early inhibition of protein synthesis by cisplatin may arise from disruption of this region. In contrast, mitochondrial dysfunction and swelling were late-stage events, and are therefore unlikely to be the primary targets of nephrotoxic platinum compounds.

Animals↗
PubMed1995

Cytotoxicity of mercury compounds in LLC-PK1, MDCK and human proximal tubular cells.

Six mercury compounds [HgCl2 (MC), Hg(CH3COO)2 (MA), Hg(NO3)2 (MN), C2H5HgSC6H4COONa (EMT), C6H5HgOCOCH3 (PMA) and CH3CIHg (MMC)] were studied using two kidney cell lines (MDCK and LLC-PK1), primary cultures of human proximal tubular cells (hPTC) and nonrenal cell lines (SAOS and Hep G2). Cell damage was measured with four different tests: neutral red uptake, mitochondrial dehydrogenase activity (MTT conversion), thymidine incorporation and protein content. Relative toxicity was established by the determination of the concentration of test compound inducing a 50% reduction of the parameter considered (EC50 value). Two groups could be distinguished: PMA, EMT and MMC are one order of magnitude more toxic than MC, MN and MA. Cellular uptake was measured by the HPLC-hybrid generation AAS after 24 hours treatment with 1.5 microM MC, MMC, PMA or EMT in MDCK cells, revealing Hg concentrations of 42.8 +/- 2.5 ng/mg protein for MC, 596.9 +/- 87.8 ng/mg protein for MMC, 269.8 +/- 75.7 ng/mg protein for PMA and of 115.9 +/- 25.2 ng/mg protein for EMT. Cytotoxicity was positively correlated with cellular uptake. The effect of the cellular GSH content on the toxicity of mercury was studied using the GSH synthesis inhibitor L-buthionine sulfoximine (BSO). In all cases an enhanced cytotoxicity was observed after BSO treatment. 2-Oxo-4-thiazolidine carboxylic acid (OTC) was used as a substrate for the GSH synthesis. Although OTC did not enhance the GSH content, the cytotoxicity of MC, MN and MA decreased significantly, no changes were observed for the other mercurials.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2003

Staurosporine-induced apoptosis and hydrogen peroxide-induced necrosis in two human breast cell lines.

The use of apoptosis-inducing agents in the treatment of malignant cancer is increasingly being considered as a therapeutic approach. In this study, the induction of apoptosis and necrosis was examined in terms of temporal dose responses, comparing a malignant and nonmalignant breast cell line. Staurosporine (SSP)-induced apoptosis and H(2)O(2)-induced necrosis were evaluated by two cytotoxicity assays, neutral red (NR) and methyl-thiazolyl tertrazolium (MTT), in comparison with a differential dye uptake assay, using Hoechst33342/propidium iodide (Hoechst/PI). Confirmatory morphological assessment was also performed by routine resin histology and transmission electron microscopy. Cell viability was assessed over a 0.5-48 h time course. In nonmalignant HBL-100 cells, 50 nM SSP induced 100% apoptosis after a 48 h exposure, while the same exposure to SSP caused only 4% apoptosis in metastatic T47D cells. Although complete apoptosis of both cell lines was induced by 50 microM SSP, this effect was delayed in T47D (24 h) compared with HBL-100 (4 h). Results also showed that neither MTT or NR can distinguish between the modes of cell death, nor detect the early onset of apoptosis revealed by Hoechst/PI.

Apoptosis↗
PubMed2001

Orphenadrine prevents 3-nitropropionic acid-induced neurotoxicity in vitro and in vivo.

1. Previous studies indicate that 3-nitropropionic acid (3-NPA) neurotoxicity involves the excitotoxic activation of N-methyl-D-aspartate (NMDA) receptors. Thus, we examined the effect of orphenadrine (an anticholinergic drug with NMDA receptor antagonist properties) on 3-NPA neurotoxicity in both cultured rat cerebellar granule cells (CGCs) and in rats. 2. Orphenadrine protected CGCs from 3-NPA-induced mortality, as assessed by both the neutral red viability assay and laser scanning cytometry, using propidium iodide staining. 3. For rats, two indirect markers of neuronal damage were used: the binding of [(3)H]-PK 11195 to the peripheral-type benzodiazepine receptor (PBR), a microglial marker, and expression of the 27 kD heat-shock protein (HSP27), a marker of activated astroglia. Systemic administration of 3-NPA (30 mg kg(-1) per day for 3 days) induced a 170% increase in [(3)H]-PK 11195 binding, and expression of HSP27. 4. Both the increase in [(3)H]-PK 11195 and HSP 27 expression were prevented by previous administration of 30 mg kg(-1) per day of orphenadrine for 3 days. Lower doses (10 and 20 mg kg(-1)) had no protective effect. Orphenadrine also reduced 3-NPA-induced mortality in a dose-dependent manner. 5. We propose that orphenadrine or orphenadrine-like drugs could be used to treat neurodegenerative disorders mediated by overactivation of NMDA receptors.

Animals↗
PubMed2005

Immunomodulatory and antitumour effects of an exopolysaccharide fraction from cultivated Cordyceps sinensis (Chinese caterpillar fungus) on tumour-bearing mice.

Cordyceps sinensis (Chinese caterpillar fungus) is a fungus parasitic on the larvae of Lepidoptera and has been considered to be a precious tonic food and herbal medicine since ancient times in China. Recently, some fungal strains have been isolated from the fruiting bodies of wild C. sinensis, and some of them have been reported to show the same properties as the natural product. In the present study, an EPSF (exopolysaccharide fraction) was prepared from cultivated C. sinensis and its effects on B16 melanoma-bearing mice were investigated. Three doses of EPSF were intraperitoneally administered every 2 days after the day of tumour-cell injection. The experiment was terminated on day 28. Phagocytosis of peritoneal macrophages and proliferation of spleen and thymus lymphocytes were assayed. The tumour metastatic foci on the lung and liver surface were checked. The expression of oncoprotein Bcl-2 in livers and lungs was assayed by a immunohistochemical method. The results showed that EPSF significantly enhanced the Neutral Red uptake capacity of peritoneal macrophages (60 mg/kg, P<0.01; 120 mg/kg, P<0.001) and spleen lymphocyte proliferation (60 mg/kg, P<0.05; 120 mg/kg, P<0.001) in B16-bearing mouse. The metastasis of B16 melanoma cells to lungs (120 mg/kg) and livers (30, 60 and 120 mg/kg) was significantly inhibited by EPSF. Moreover, EPSF decreased the levels of Bcl-2 in the lungs (120 mg/kg) and livers (30, 60 and 120 mg/kg). These results suggest that EPSF has immunomodulatory function and antitumour activity.

Animals↗