ABSTRACT
Ovarian reserve and fertility are reduced by aging and a poor energy balance. To date, the relationships of high energy accumulation and aging with the ovarian reserve have not been elucidated. Here, the effects of obesity on the aging ovarian reserve were evaluated in a leptin-deficient (ob/ob) mouse model. Abnormal estrous cyclicity appeared as early as 6 weeks and worsened with aging. The blood level patterns of 17ß-estradiol (E2), testosterone (T), and progesterone (P4) with aging were similar between lean and ob/ob mice. The blood level of E2 but not P4 or T was similar at 24 weeks. Many more atretic follicles but fewer corpora lutea were observed in ob/ob mice than in lean mice within all age groups. Anti-Müllerian hormone (Amh) mRNA levels were similar between genotypes. Dazl, Stra8, and ZP3 mRNAs were highly expressed in ob/ob mice after 12 weeks. Sohlh1 and Ybx2 mRNAs were highly expressed at 24 weeks in ob/ob compared with lean mice. In addition, SOHLH1-positive primordial follicle counts were significantly increased in ob/ob mice at 24 weeks. The proportions of AMH-positive secondary and small antral follicles were similar between genotypes. Together, these results show that the ovarian reserve lasts longer in ob/ob mice than in lean mice, suggesting that the loss of normal physiological or physical status causes decreased fertility at a young age in ob/ob mice and that an increase in adipocytes without leptin, as in ob/ob mice, can improve the ovarian reserve. Such knowledge can be applied to understanding reproductive dysfunction.
ABSTRACT
BACKGROUND/AIM: (-)-Epicatechin is a major constituent of Bulnesia sarmienti, which is known to possess anticancer properties. Here we report that (-)-epicatechin isolated from B. Sarmienti inhibited growth and induced apoptosis of SW480 human colon cancer cells. MATERIALS AND METHODS: Cells were treated with different concentrations (0, 25, 50, and 100 µmol/ml) of (-)-epicatechin. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, 40,6-diamidine-20-phenylindole dihydrochloride (DAPI) staining, colony-forming assay, DNA fragmentation analysis, reverse transcription-polymerase chain reaction (RT-PCR), annexin V-fluorescein isothiocyanate (FITC) staining, and immunoblot analyses were then carried out. RESULTS: (-)-Epicatechin was found to have cytotoxic activity, and cells treated with this compound had fragmented nuclei, fragmented DNA, and underwent apoptosis. mRNA and protein expression levels of BCL2-associated X protein (BAX) and p53 were up-regulated and those of B-cell lymphoma-2 (BCL2) were down-regulated, while p21 mRNA levels were significantly increased in cells treated with (-)-epicatechin in a concentration-dependent manner. CONCLUSION: (-)-Epicatechin from B. Sarmienti inhibited colon cancer cell growth and induced apoptosis.
Subject(s)
Apoptosis/drug effects , Catechin/pharmacology , Colonic Neoplasms/drug therapy , Colonic Neoplasms/pathology , Antineoplastic Agents, Phytogenic/isolation & purification , Antineoplastic Agents, Phytogenic/pharmacology , Catechin/isolation & purification , Cell Line, Tumor , Colonic Neoplasms/genetics , Colonic Neoplasms/metabolism , Fluorescent Dyes/chemistry , Humans , Indoles/chemistry , Microscopy, Fluorescence , Proto-Oncogene Proteins c-bcl-2/biosynthesis , Proto-Oncogene Proteins c-bcl-2/genetics , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Stereoisomerism , Tumor Suppressor Protein p53/biosynthesis , Tumor Suppressor Protein p53/genetics , Zygophyllaceae/chemistry , bcl-2-Associated X Protein/biosynthesis , bcl-2-Associated X Protein/geneticsABSTRACT
Cordyceps militaris (CM) is an insect-borne fungus that has been used in traditional Chinese medicine because of its wide range of pharmacological activities. In this paper, we studied CM grown on germinated soybean (GSC) and investigated the possible mechanisms underlying antiproliferative effect of GSC on HT-29 human colon cancer cells. In comparison with CM extracts and germinated soybean (GS) BuOH extracts, BuOH extracts of GSC showed remarkable inhibitory and antiproliferative effects on HT-29 colon cancer cells. After GSC treatment, HT-29 cells became smaller and irregular in shape. High G2/M phase cell populations were observed in the GSC-treated group. The levels of cyclin B1 and Cdc25 in the GSC-treated group were lower than those in the control group. These findings suggest that GSC BuOH extracts might act as an effective anti-proliferative agent by inducing G2/M cell cycle arrest in colon cancer cells.
ABSTRACT
BACKGROUND: Ionizing radiation is used to treat many of cancers, however, it also produces unwanted side effect on normal tissues, such as radiodermatitis. We previously established an animal model for radiodermatitis, and found that X-ray irradiation induced the expression of ID3 in hairless mouse skin by cDNA microarray. OBJECTIVE: The aim of this study is to investigate the functional role of ID3 in X-ray irradiated keratinocytes. METHODS: Immunohistochemistry, RT-PCR and Western blot were performed to demonstrate the ID3 induction by X-ray irradiation. HaCaT keratinocytes were transduced with the recombinant adenovirus expressing HA-ID3, and then effects on apoptosis were analyzed. RESULTS: X-ray irradiation increased markedly the ID3 protein level in epidermis of mouse skin. X-ray irradiation also induced the expression of ID3 in HaCaT keratinocytes cultured in vitro, at both mRNA and protein levels. When ID3 was overexpressed by recombinant adenovirus, apoptosis of keratinocytes were induced even in the absence of X-ray irradiation. Furthermore, overexpression of ID3 sensitized X-ray-induced apoptosis. Interestingly, X-ray irradiation significantly reduced the endogenous ß-catenin level, which was related with induction of apoptosis. Similarly, overexpression of ID3 led to remarkable reduction in ß-catenin level. CONCLUSION: These results suggest that ID3 plays a role as an apoptosis inducer in response to X-ray irradiation via the regulation of endogenous ß-catenin level.
Subject(s)
Apoptosis/physiology , Apoptosis/radiation effects , Inhibitor of Differentiation Proteins/metabolism , Keratinocytes/metabolism , Keratinocytes/radiation effects , Neoplasm Proteins/metabolism , beta Catenin/metabolism , Animals , Apoptosis/genetics , Base Sequence , Cell Line , DNA Primers/genetics , Humans , Immunohistochemistry , Inhibitor of Differentiation Proteins/genetics , Keratinocytes/cytology , Male , Mice , Mice, Hairless , Neoplasm Proteins/genetics , RNA, Messenger/genetics , RNA, Messenger/metabolismABSTRACT
The effects of Bulnesia sarmienti (BS) aqueous extract on the cell growth of A549 cell lines were investigated. BS has strong cytotoxic activity on the A549 cell lines (IC(50); less than 100 microg/mL) in MTT assay. HPLC confirmed that BS contains catechins as major compound. Cell cycle analysis by flow cytometry indicated that BS arrested the cell cycle in the sub-G(1) phase. BS induced DNA fragmentation, and increased the expression of the p53 protein in immunoblot analysis. These results indicated that the anticancer effect of BS was mediated via the process of apoptosis and growth-inhibition.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , DNA Fragmentation/drug effects , G1 Phase/drug effects , Magnoliopsida/chemistry , Plant Bark/chemistry , Plant Extracts/pharmacology , Animals , Antineoplastic Agents, Phytogenic/chemistry , Catechin/chemistry , Catechin/pharmacology , Cell Line, Tumor , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Humans , Mice , Plant Extracts/chemistry , Tumor Suppressor Protein p53/metabolismABSTRACT
The antiobesity effect of wild ginseng (WG; Panax ginseng C.A. Meyer) in male obese leptin-deficient (B6.V-Lepob, 'ob/ob') mice was evaluated. WG was administered orally to mice at doses of 100 mg/kg and 200 mg/kg daily for 4 weeks. The WG-treated ob/ob mice showed a loss of body weight and a decrease in blood glucose levels compared with control mice. WG regulated the mRNA expression level especially, it increased peroxisome proliferators-activated receptors-gamma (PPAR-gamma) and lipoprotein lipase (LPL) in adipose tissue, as well as glucose transporter 4 (GLUT4) and insulin receptor (IR) in the skeletal muscle and liver. Taken together, these results suggest that WG may play a vital role in the antiobesity effect in ob/ob mice; this has importance in insulin sensitivity. This may prove to be of clinical importance in improving the management of obesity and related metabolic syndromes.
Subject(s)
Anti-Obesity Agents/pharmacology , Glucose Transporter Type 4/metabolism , Lipoprotein Lipase/metabolism , PPAR gamma/metabolism , Panax/chemistry , Adipose Tissue/drug effects , Animals , Blood Glucose/drug effects , Body Weight/drug effects , Leptin/deficiency , Liver/metabolism , Male , Mice , Muscle, Skeletal/metabolism , Obesity/drug therapy , Receptor, Insulin/metabolismABSTRACT
Bulnesia sarmienti (BS), a traditional South American herbal medicine native to Gran Chaco, has been used to treat various human ailments. The effects of BS aqueous extract (100, 200, and 400 microg/ml) on H460 cell lines were investigated. High-performance liquid chromatography (HPLC) confirmed that BS contains catechins as major compound. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell cycle analysis, DNA fragmentation, apoptosis, and immunoblot analysis on cells were carried out. BS has strong cytotoxic activity on the H460 cell lines (IC50; less than 100 microg/ml) in MTT assay. Flow cytometry indicated that BS arrested the cell cycle in the sub-G1 phase. When BS was treated on H460 cells, DNA fragmentation was increased, and early apoptotic cells were shown to be positive by annexin V staining. Also, the expressions of the p53 and Bax were increased and Bcl-2 protein was downregulated with BS treatment. These results indicated that the BS has anticancer activity on H460 cells and BS may be useful in future therapeutic applications for developing anticancer agents.
