The root of Adenophora triphylla is a highly valued medicinal resource that is used to prevent human obesity, cancer, and inflammation, whereas young leaves or sprouts of A. triphylla are used as food ingredients. In this study, we compared the antioxidant and anticancer activities of 70% ethanol extracts of A. triphylla roots and leaves. The leaf extract exhibited stronger 2,2-diphenyl-1-picrylhydrazyl (DPPH)-radical scavenging activity, reducing power, and oxygen radical absorbance capacity (ORAC) than the root extract. Furthermore, the leaf extract was observed to be a potent source of anticancer compounds that were effective against A549 (lung cancer), LNcaP (prostate cancer), SKOV3 (ovarian cancer), and Caco-2 (colorectal cancer) cells. These results indicate that not only the roots but also the leaves of A. triphylla can serve as valuable sources of functional materials in the pharmaceutical industry.
Seo, Kil Woong;Kim, Chong Sup;Park, Chang Sik;Lee, Kyu Seung
Korean Journal of Agricultural Science
/
v.17
no.1
/
pp.34-44
/
1990
The study was carried out to elucidate the effects of ovarian function on the thyroid gland, adrenal gland and uterus in female rats. One hundred and forty-four mature female rats were allotted into the three groups ; ovariectomized group, estradiol treated group and intact control group. The ovaries of 48 heads of rats were completely removed. Forty eight heads of rats were administered with $200{\mu}g$ of estradiol benzoate every 48 hours. Serum estradiol-$17{\beta}$ and progesterone levels were determined with radioimmunoassay method at 3, 6, 12, 24 hours and 5, 10, 15 days after treatment. The rats were necropsied to measure weights of thyroid gland, adrenal gland and uterus and to examine the histological changes in the organs. The results obtained were as follows ; 1. Serum estradiol-$17{\beta}$ levels were rapidly decreased below 27.20pg/ml 18 hours after ovariectomy. In estradiol treated rats the levels were rapidly increased 18 hours after treatment, but thereafter slowly decreased. The significant differences in the estradiol level were found between the group at every observation time. 2. Serum progesterone levels were significantly decreased after ovariectomy and estradiol injection. The lowest level was found in the group of ovariectomized rats. 3. The weights of thyroid glands decreased in ovariectomized rats rather than in intact rats 5 days after treatment. The weights tended to increase after estradiol injection but significant differences between the groups were seen on 10th and 15th days. 4. In the histological findings of thyroid glands, follicular epithelial cells were changed to be squamous 5 days after ovariectomy and accompanied pyknosis 10 days and karyorrhexis 15 days after ovariectomy. On the contrary follicular epithelial cells were changed to be columnar with hypertrophy 10 days after estradiol injection. 5. The significant differences in adrenal gland weights were recognized between all the groups 5 and 15 days after treatment in ovariectomized rats were lighter than intact rats and the adrenal gland weights were rather heavier in estradiol treated rats. 6. The days after ovariectomy the adrenal glands were atrophied accompanying with pyknosis in the cortical cells of zona fasciculata. The cells in zona fasciculata and zona reticularis started to hypertrophy 5 days after estradiol injection, but no changes were found in the zona glomerulosa of adrenal cortex and in the adrenal medulla. 7. The significant differences in uterus weights were recognized between the groups at each observation time. After ovariectomy the uterus weights decreased rapidly but after estradiol injection they increased rapidly. 8. Through histological examination of uterus, the atrophy and degeneration started to occur in endometrium and lamina propria 12 hours after ovariectomy, and in myometrium one day after ovariectomy, and the changes progressed rapidly after that. On the contrary, the myometrium was proliferated and hypertrophied from 12 hours after estradiol-$17{\beta}$ injection.
Outlines for plasma $estradiol-17\beta$, components, electrophoretic patterns, and ultrastructural changes were obtained in female rainbow trout (Oncorhynchus mykiss) during the seasonal reproductive cycles. Plasma $estradiol-17\beta$ under the natural conditions, exhibited distinct seasonal variation, peaking very late in vitellogenic season during September, decreasing gradually the halt of spawning in December, and ultimately falling during the early stages of seasonal ovarian recrudescence in February and March. This change in $estradiol-17\beta$ appeared to stimulate vitellogenin production as evidenced by increases in plasma calcium, phosphorus, glucose, albumin and total protein levels. The electrophoretic patterns of late maturing or spawning oocytes were stained more intensively than those of late perinucleolus oocytes (molecular weights of approximately 70,000 and 200,000). Two protein bands were found in the SDS-PAGE separation, coincident with the $estradiol-17\beta$ hormone peak. Gonadosomatic indices (GSI) significantly increased from October to January, and showed the highest peak in January, coinciding with the numerically abrupt increase of ripe ova in female. A positive correlation (r=0.701, p<0.01) was established between plasma $estradiol-17\beta$ levels and the gonadosomatic index during the prespawning. The highest level of hepatosomatic index (HSI) observed in December. During the breeding season (December), the gonadotropes were large and filled with GTH-containing inclusions such as granules and globules. The vitellogenic phase began as late perinurleolus oocytes became transformed into early maturing oocytes through the accumulation of yolk, and oocytes reached the late maturing stages as the ooplasm was completely packed with yolk. Marked ultrastructural changed in the granulosa cells during nuclear migration involve the dilation of the rough endoplasmic reticulum and the appearance of the rod-shaped mitochondria with tubular cristae. Microvilli (finger-like projections), from the zona radiata and from the oocyte grew, and made contact with each other in the pore canals of the zona radials during vitellogenesis, but were withdrawn as the zona radiata became more compact and devoid of pore canals during oocyte maturation. The zona radiata grew to a tripartite structure such as an outer thin homogeneous layer, and two inner thick helicoidal layers (zona radials interna and zona radiata externa). Under the normal conditions, the ovarian follicle influenced the histological development and periodical secretion of the hormones , sufficient for a oogenesis and gonadal steroid production.
Cho Dong-Jae;Park Cheol-Hong;Yang Hyunwon;Park Joo-Hyun;Yun Jeong-Mi;Kim Sei-Kwang;Yoon Yong-Dal
Development and Reproduction
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v.8
no.1
/
pp.27-33
/
2004
To investigate the mechanism of germ cell death in postnatal stage of mouse, the involvement of apoptotic executioners, caspase-3 and caspase-activated DNase(CAD), and apoptotic initiators, Bax Fas and Fas ligand, in the germ cell death has been studied. Immune-labels of active caspase-3 and CAD were located in TUNEL-positive, apoptotic, oocytes as well as normal oocytes of primary or secondary follicles. CAD immune-labels were also detected in the nucleus of TUNEL-positive oocytes. Most of oocytes showing positive immune-labeling of active caspase-3 or CAD had vacuoles in their cytoplasm, which is the morphological characteristic of oocyte during folliclar atresia. Bax immune-stains were detected in the atretic oocytes which showed the vacuole in their cytoplasm. Positive immune-labels for Fas ligand was localized in TUNEL-positive or atretic oocytes. Presence of immunoreactivity of active caspase-3 and CAD in TUNEL-positive germ cells implicate that active raspase-3 and CAD might play a role in germ cell apoptosis during early development of mouse ovarian follicle. Immunohistochemical localization of Bax and Fas ligand in TUNEL-positive oocytes suggests that these might be the most plausible modulator of oocyte apoptosis.
Background: Fascin is an actin-bundling protein that plays an important role in cellular motility. Fascin is normally expressed in the neuronal and mesenchymal cells and its expression is low or absent in the epithelia. However, an overexpression of fascin has been linked to the invasive behavior of some neoplasms such as breast, stomach and ovarian tumors. In this study, we evaluated the expression of fascin and its prognostic significance in stage I non-small cell lung cancer (NSCLC). Methods: Immunohistochemical staining for fascin was performed on the paraffin-embeded tissue sections of 81 cases of resected NSCLC. Staining of more than 5% of the tumor cells was recorded as positive immunoreactivity. Results: Fascin expression was seen in 73% (59/81) of the cases and this was more frequently seen in squamous cell carcinoma than in adenocarcinoma (93% vs 42%). There were no significant correlations of fascin immunoreactivity with tumor recurrence and overall survival. Conclusion: The expression rate of fascin was relatively high in NSCLC, but this was without prognostic significance. The exact clinical role of fascin should be defined through further investigations.
Despite progress in elucidating mechanisms associated with colorectal cancer and improvement of treatment methods, it remains a frequent cause of death worldwide. New and more effective therapies are therefore urgently needed. Recent studies have shown that immunogenicity of whole ovarian tumor cells and subsequent T cell response were potentiated by oxidation modification with hypochlorous acid (HOCl) in vitro and ex vivo. These results prompted us to investigate the protective antitumor response with an HOCl treated CT26 colorectal cancer cell vaccine in an in vivo mouse model. Administration of HOCl modified vaccine triggered robust antitumor immunity to autologous tumor cells in mice and prolonged survival period significantly. In addition, increased necrosis and apoptosis were found in tumor tissue from the oxidation group. Interestingly, ELISPOT assays showed that specific T cell responses were not elicited in response to the immunizing cellular antigen, in contrast to raising sera antibody titer and antibody binding activity shown by ELISA assay and flow cytometry. Further evaluation of the mechanisms underlying HOCl modified vaccine mediated humoral immunity highlighted the role of antibody-dependent cell-mediated cytotoxicity. These results combined with previous studies suggest that HOCl oxidation modified whole cell vaccine has wide applicability as a cancer vaccine because it can target both T cell- and B cell-specific responses. It may thus represent a promising approach for the immunotherapy of colorectal cancer.
In mammalian ovary, major portion(>99%) of ovarian follicles undergo atresia. Recent studies have shown that this phenomenon is mediated via GC apoptosis. Ceramide, a product of sphingomyelin hydrolysis, has been proposed as a novel lipid second messenger with specific roles in mediating antiproliferative responses including apoptosis and cell cycle arrest. In the present study, we have examined the effect of ceramide on apoptotic cell death of GC in vitro. GCs were harvested by squeezing the antral follicles from the immature mice (3-4 weeks) and cultured in MEM medium with 10% fetal bovine serum. The cells were treated with various concentrations of ceramide (0 to 50 \mu M)and cultured up to 24 h.Cell death was determined by MTT cell viability assay and apoptosis was examined by acridine orange staining, in situ 3'-end labeling(TUNEL), and flow cytometry. Ceramid treatment induced apoptotic cell death of GC in a time- and a dose-dependent manner. Results of flow cytometric analysis showed that creamide-induced cell death was mostly confined to the $G_{0}$/$G_{1}$ cells. these results provide an evidence for ceramide as a lipid second messenger of apoptosis in mouse GC.
When mammalian oocytes undergo maturation, cumulus cells surrounding the oocyte exhibit remodeling of their structure known as cumulus expansion. Many molecules including hyaluronic acid participate in this remodeling. The present study aimed to investigate a possible existence of matrix metalloproteinases(MMPs) in the extracellular matrix(ECM) of human oocyte-cumulus complex. ECM was extracted from the human oocyte-cumulus complex. Gelatin gel zymogram of ECM exhibited 7 gelatinases having molecular weight of 300kDa, 240kDa, 200kDa, 180kDa, 116kDa, 97kDa, and 84kDa. This gelatinase profile was very different from that of ovarian mural granulosa cell extract or white blood cell extract, indicating that the oocyte-cumulus complex donating ECM was free from other than cumulus cells. When ethylenediaminetetraacetic acid or 1', 10'-phenanthroline was added to the reaction buffer during zymographic development, almost gelatinase activities were abolished, suggesting that they were MMPs. Following incubation of ECM in the presence of aminophenylmercuric acetate, an activator of proMMPs, 4 gelatinases of 240kDa, 180kDa, 97kDa, and 84kDa disappeared with the concomitant appearance of 80kDa, 65kDa, and 60kDa gelatinases. Based upon these observation, it is suggested that ECM of the human oocyte-cumulus complex consists of gelatinases, presumed to be MMP-2 and MMP-9 isoforms.
Zhang, Yong-Chun;Jiang, Gang;Gao, Han;Liu, Hua-Min;Liang, Jun
Asian Pacific Journal of Cancer Prevention
/
v.15
no.5
/
pp.2353-2358
/
2014
Purpose: We aimed to detect the expression of HIF-1${\alpha}$, VEGF, HPSE-1 and CD31 in SKOV3 xenografts in nude mice treated with different doses of ionizing radiation, trying to explore the possible mechanism of hypoxia and radioresistance. Methods: Nude mice bearing SKOV3 xenografts were randomly divided into 4 groups: Group A (control group, no ionizing radiation), Group B (treated with low dose of ionizing radiation: 50cGy), Group C (treated with high dose of ionizing radiation: 300cGy), Group D ( combined ionizing radiation, treated with ionizing radiation from low dose to high dose : 50cGy first and 300cGy after 6h interval). The mRNA levels of HIF-1 and VEGF in each group were detected by real time polymerase chain reaction, while HPSE-1 expression was measured by ELISA. The microvessel density (MVD) and hypoxic cells were determined through immunohistochemical (IHC) staining of CD31 and HIF-1a. Results: Significant differences of HIF-1${\alpha}$ mRNA level could be found among the 4 groups (F=74.164, P<0.001): Group C>Group A>Group D> Group B. The mRNA level of VEGF in Group C was significantly higher than in the other three groups (t=-5.267, P=0.000), while no significant difference was observed among Group A, B and D (t=1.528, 1.588; P=0.205, 0.222). In addition, the MVD was shown to be the highest in Group C (t=6.253, P=0.000), whereas the HPSE-1 level in Group A was lower than in Group B (t=14.066, P=0.000) and higher than in Group C (t=-21.919, P=0.000), and similar with Group D (t=-2.066, P=0.058). Through IHC staining of HIF-1a, the expression of hypoxic cells in Group A was (++), Group B was (+), Group C was (+++) and Group D was (+). Conclusion: Ionizing radiation with lowerdoses might improve tumor hypoxia through inhibiting the expression of HIF-1 and HPSE-1, whereas higherdoses worsen tumor hypoxic conditions by up-regulating HIF-1${\alpha}$, HPSE-1, VEGF and CD31 levels. A protocol of low-dose ionizing radiation followed by a high-dose irradiation might at least partly improve tumor hypoxia and enhance radiosensitivity.
Purpose : This study was designed to investigate the effects of Jeongkyeong-Tang(JKT) on the polycystic ovary(PCO) induced by estradiol valerate(EV) in rats. Methods : PCO was induced by single intramuscular injection with EV(4mg) in female rats. Normal group(n=8) were injected with sesame oil and orally administrated distilled water for sixty days. PCO control group(n=8) were injected with EV and orally administrated distilled water for sixty days. JKT treated group(n=8) were injected with EV and orally administrated JKT for sixty days. Then we measured weights of body, ovaries and adrenal glands, and measured content of serum estrogen. The histomorphometrical changes of ovaries were also evaluated. The expressions of nerve growth factor(NGF) were analyzed in the central nervous system, adrenal glands and ovaries by immunohistochemistry. Results : - The weights(mg) of ovaries in JKT treated group (69.7${\pm}$6.7) were significantly increased( p<0.001) compared with PCO control group(46.7${\pm}$12.2). - The numbers of secondary follicles in JKT treated group(4.00${\pm}$l.31) were significantly increased(p <0.05) compared with PCO control group(2.25${\pm}$1.39). - The numbers of mature follicles in JKT treated group(5.50${\pm}$1.51) were significantly increased(p<0.01) compared with PCO control group(2.88${\pm}$1.13). - The numbers of atretic follicles in JKT treated group(2.75${\pm}$l.16) were significantly decreased(p<0.001) compared with PCO control group(6.88${\pm}$2.03). - The numbers of corpora lutea in JKT treated group(4.13${\pm}$1.46) were significantly increased(p<0.01) compared with PCO control group(2.13${\pm}$1.46). - The contents(pg/ml) of serum estrogen in JKT treated group(115.18${\pm}$18.29) were significantly decreased(P<0.01) compared with PCO control group(153.06${\pm}$29.47). - The expressions of NGF-immunoreactive cells in the ovarian granulosa cells in JKT treated group were lesser observed than PCO control group. Conclusion : From the above results, we concluded that Jeongkyeong-Tang has inhibitory effect on the development of EV-induced polycystic ovary. And it's effect may be related with decreased NGF activities in the ovary.
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