Effects of herbal medicine on human uterine tumor-bearing nude mice.

AIM: Uterine leiomyomas are the most common benign uterine neoplasms associated with significant morbidity. Herbal formulas capable of restoring yin-yang balance by dispersing blood stasis may be useful for managing fibroid symptoms. MATERIALS AND METHODS: In this study, the antitumor properties of three herbs viz., Trogopterus xanthipes Milen-Edwards, Paeonia lactiflora Pallas, and Ulmus davidiana Planch were evaluated in nude mice injected intravenously with human malignant myomas. Tumor fragments were xenografted subcutaneously through a flank incision in female mice. The mice entered the study for 8 weeks when their tumors reached the threshold volume (260 mm3). The mice were randomly allocated to receive subcutaneous injections of normal saline (Group 1; negative control), P. lactiflora Pallas (Group 2), U. davidiana Planch (Group 3), T. xanthipes Milen-Edwards (Group 4), and intravenous injections of paclitaxel (Group 5; positive control). The weight and tumor volume were measured, followed by histopathology. RESULTS: A few cases of abdominal distention and death were observed in the negative control group. Furthermore, a considerable enlargement of the liver and spleen was observed in the negative control group at autopsy with a gradual increase in body weight during the experiment. The mean tumor volume which increased in negative control mice reduced in mice treated with herbal remedies or paclitaxel from day 14 onwards (P < 0.05). The degree of necrosis and apoptosis induction from herbal treatments was similar to that of paclitaxel. CONCLUSION: Collectively, three herbs viz., T. xanthipes Milen-Edwards, P. lactiflora Pallas, and U. davidiana Planch were able to induce necrosis and apoptosis of uterine leiomyoma cells, proving antitumor properties against uterine fibroids.

Effects of Dietary Supplementation with Astaxanthin on Histamine Induced Lesions in the Gizzard and Proventriculus of Broiler Chicks.

Astaxanthin (ASX) is a xanthophyll pigment isolated from crustaceans and salmonids. Owing to its powerful antioxidant activity, ASX has been reported to have the potential to protect against gastric ulcers and a variety of other illnesses. Histamine (His) is a dietary factor that causes gastric erosion and ulceration in young chicks. In this study, we examined whether ASX had protective effects on dietary histamine-induced lesions in the gizzard and proventriculus of broiler chickens. Four experimental treatment groups were planned: basal diet (BD), BD+His, BD+ASX, and BD+ASX+His, with four chicks (5 days old) in each group and three replications (i.e., a total of 12 chicks per group). The BD was supplemented with either 0.4% His or 100 ppm ASX. The birds were fed ad libitum for 3 weeks, and diets contained no antimicrobial compounds. Supplementing the diet with His significantly decreased body weight gain, but increased the weights of the gizzard and proventriculus of the chicks as compared with those of chicks in the BD group (p<0.05). ASX did not affect His-dependent changes in chick body weight or weights of the gizzard and proventriculus. The loss of gastric glands in the proventriculus, which was observed in His-treated chicks, was not prevented by ASX administration. The frequency of proventricular ulceration, however, was lowered by treatment with ASX, without significant differences between the two supplementation levels. In conclusion, our data showed that ASX might be helpful for alleviating structural damage to the digestive system in poultry under certain stressful conditions.

Effect of Lactobacillus acidophilus KFRI342 on the development of chemically induced precancerous growths in the rat colon.

Lactobacillus acidophilus KFRI342, isolated from the Korean traditional food kimchi, was investigated for its suitability as a dietary probiotic. The effects of L. acidophilus KFRI342 on the development of chemically induced (1,2-dimethylhydrazine; DMH) precancerous cytological changes of the colon were investigated in rats. Forty-five male F344 rats were randomly divided into three dietary groups. The control group received a high-fat diet (HF), a second group received a high-fat diet containing the carcinogen (HFC), and a final group received a high-fat diet containing the carcinogen and L. acidophilus KFRI342 (HFCL). L. acidophilus KFRI342 was administered orally three times per week at 2×10(9) c.f.u. ml(-1). L. acidophilus KFRI342 treatments decreased the number of Escherichia coli in faecal samples, the enzyme activities of ß-glucuronidase and ß-glucosidase, and plasma triglyceride concentration compared to the HF and HFC treatments (P<0.05). L. acidophilus KFRI342 consumption also decreased the ratio of aberrant crypts to aberrant crypt foci incidence and the number of aberrant crypts in HFCL rats. Therefore, L. acidophilus showed potential probiotic activity as an inhibitor of DMH-induced symptoms in live rats. Our in vivo studies indicate that L. acidophilus from kimchi may be suitable as a probiotic for human use.

Cytotoxicity of 5-fluorouracil: Effect on endothelial differentiation via cell cycle inhibition in mouse embryonic stem cells.

Embryonic stem cells (ESCs) are known to characteristics for pluripotency and self-renewal, but the precise mechanisms of ES-derived cells to specific toxicants have not been determined. Here, we evaluated the cytotoxicity of 5-fluorouracil (5-FU) and see its effect on cell viability, proliferation, and differentiation in mouse ESC-derived endothelial differentiation. Mouse ESCs were exposed to 5-FU (10 microM) and combined with probucol (50 microM) for 24h, which is an antagonist of 5-FU. Changes in gene expression as a result of 5-FU exposure in mouse ESC-derived endothelial precursor cells (ES-EPCs) were assessed using an oligonucleotide microarray (AB1700). The expression of Oct-4 was decreased during the differentiation of mouse ESCs into endothelial cells; otherwise, the expression of PECAM was increased. Mouse ES-EPCs were shown to have a decrease in viability (49.8%) and PECAM expression, and induce G1/S phase (31.1%/60.6%) when compared with/without treatment of 5-FU. Expression of cell cycle-related proteins was increased in endothelial precursor cells exposed to 5-FU without probucol treatment. From theses results suggest that 5-FU inhibit endothelial differentiation as well as inducing the G1/S phase arrest. We propose that mouse ES-EPCs might be a useful tool for screening the cytotoxicity of compounds in endothelial cells.

Differentiation of endothelial cells derived from mouse embryoid bodies: a possible in vitro vasculogenesis model.

Mouse embryonic stem cells (mES cells), which are pluripotent and self-renewal cells, are derived from the inner cell mass of mouse blastocysts. The objective of this study was to construct more efficient mES cell-derived embryoid bodies (EBs) for use as a vasculogenesis model and as an in vitro vascular toxicity testing model. EBs were formed for 3 days using hanging drop cultures and plated on gelatin-coated plates in endothelial growth medium-2 (EGM-2) to promote vascular development. The differentiation of mES cell-derived EBs was confirmed by reverse transcription-polymerase chain reaction (RT-PCR), immunocytochemistry, and flow cytometry within 7 days after plating EBs. The mRNA and protein expressions of vascular endothelial growth factor receptors-2 (FLK-1), platelet endothelial cell adhesion molecule (PECAM), and vascular endothelial-cadherin (VE-cadherin) were observed in differentiated mES cells. When placed in matrigel, mES cell-derived endothelial like cells formed networks similar to vascular structures. mES cells were also exposed to 5-fluorouracil (5-FU), a strong inhibitor of vessel formation, and its cytotoxicity was determined using MTT assays. The inhibitory concentrations (IC50) of 5-FU for mES cells and C166 cells were 0.72 microM and 1.04 microM, respectively. These results demonstrate that mES cells can be used to study vasculogenesis and for cytotoxicity screening.

Effects of hesperetin on vessel structure formation in mouse embryonic stem (mES) cells.

OBJECTIVE: The present study investigated the effects of hesperetin on vessel structure formation in mouse embryonic stem (mES) cells with regard to whether hesperetin acts as an antioxidant or pro-oxidant. Some flavonoids enhance antioxidant systems while increasing oxidative stress in the body. METHODS: After their differentiation into endothelial-like cells for 10 d, mES cells were treated with 1 to 100 muM of hesperetin for 24 h. RESULTS: Hesperetin efficiently inhibited the formation of vessel-like tubular structures consisting of platelet-endothelial cell adhesion molecule-1-immunoreactive cells and significantly (P < 0.05) increased the generation of reactive oxygen species in a concentration-dependent manner. Although glutathione (in its reduced and oxidized forms) in mES cells was not affected by hesperetin, the 8-iso-prostaglandin F2(alpha) content was decreased. In addition, cytotoxicity-induced hesperetin was not found; lactate dehydrogenase release and cell viability were determined as an index of cell damage. CONCLUSION: Taken together, the present study shows that hesperetin inhibits vessel formation by pro-oxidant means and suggests its potential as an antiangiogenic agent.

Long-term combined administration of quercetin and daidzein inhibits quercetin-induced suppression of glutathione antioxidant defenses.

In this study, we investigated the effects of long-term administration of quercetin with or without daidzein on glutathione and the enzymes involved in its metabolism in rat liver in vivo. Male Sprague-Dawley rats were divided randomly into four groups and given oral quercetin (20 mg/day) and daidzein (20 mg/day) alone or in combination, or vehicle alone for six weeks. The serum and liver alpha-tocopherol concentrations were significantly increased following administration of quercetin and daidzein alone or in combination. Glutathione concentration and glutathione reductase activity was significantly (p < 0.05) decreased with quercetin treatment, while no such effect was observed with daidzein treatment. Interestingly, decrease in glutathione concentration and glutathione reductase activity by quercetin treatment was inhibited by combined administration of daidzein and quercetin. The malondialdehyde concentration was significantly decreased following administration of quercetin and daidzein alone or in combination. These results suggest that quercetin, but not daidzein, acts as a pro-oxidant agent by decreasing glutathione concentration and glutathione reductase activity. Interestingly, this pro-oxidant effect of quercetin was inhibited by the combined administration of quercetin and daidzein.

Anti- and prooxidant effects of chronic quercetin administration in rats.

The present study was performed to investigate the effects of chronic administration of quercetin on lipid peroxidation and glutathione concentration in rat liver. Male Sprague-Dawley rats were divided into two groups, one of which was fed a normal diet and the other a vitamin E-free diet. Each of these groups was divided further into three subgroups and treated with quercetin administered orally at either 2 or 20 mg/day or with vehicle for 4 weeks. The concentrations of alpha-tocopherol in serum and liver increased following quercetin treatment, and these increases were significantly greater in rats maintained on a vitamin E-free diet. Quercetin significantly decreased the concentration of malondialdehyde (an indicator of lipid peroxidation) in the liver and this decrease was more pronounced in vitamin E-deprived rats than in those maintained on a normal diet (55-60% and 25-35% decrease in malondialdehyde concentrations, respectively). Quercetin treatment decreased the glutathione concentration and glutathione reductase activity (40 and 34%, respectively) in the liver significantly and to a similar extent in vitamin E-deprived and -undeprived rats. Collectively, these results suggest that quercetin may act not only as an antioxidant, but also as a prooxidant in rats.