• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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• 2003.04a
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• pp.22-39
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• 2003

Biological activity of phenolic compounds in seeds and leaves of safflower (Carthamu tinctorius L.) were evaluated using several in vitro and in vivo assays. Six phenolic constituents were isolated from the seeds and identified as N-feruloylserotonia, N- (p-coumaroyl)serotonin, matairesinol, 8′-hydroxyarctigenin, acacetin 7-O-$\beta$-D-glucoside (tilianine) and acacetin. Six phenolic compounds exhibited considerable antioxidative activity, and especially two serotonins showed potent DPPH radical scavenging activity and antiperoxidative activity against rat liver microsomal lipid peroxidation induced by the hydroxyl radical generated via a Fenton-type reaction. Additionally, six phenolic compounds possessed comparable cytotoxicity against three cancer cells, Hela cell, MCF-7 and HepG2 cell, and particularly acacetin and its glycosides had the most potent cytotoxicity. Moreover, we found that feeding safflower seeds attenuated bone loss, and lowered levels of plasma and liver lipids in ovariectomized rats. Serotonins, lignans and flavones stimulated proliferation of the osteoblast-like cells in a dose-dependent manner (10$^{-15}$ ~10$^{-6}$ M), as potently as E$_2$ (17$\beta$-estradiol). Particularly, serotonins were mainly responsible for bone-protecting and lipid lowering effects in ovariectomized rats. Meanwhile, eight flavonoids, including a novel quercetin-7-O-(6"-O-acetyl)-$\beta$-D-glucopyranoside and seven kown flavonoids, luteolin quercetin, luteolin 7-O-$\beta$-D-glucopyranoside, luteolin-7-O-(6"-O-acetyl)-$\beta$-D-gluco-pyranoside, quercetin 7-O- -glucopyranoside, acacetin 7-O-$\beta$-D-glucuronide and apigenin-6-C-$\beta$-D-glucopyranosyl-8-C-$\beta$-D-glucopyranoside were first isolated and identified from safflower leaf. Among these flavonoids, luteolin-acetyl-glucoside and $\beta$quercetin- acetyl-glucoside showed potent antioxidative activities against 2-deoxyribose degradation and lipid peroxidation in rat liver microsomes. Luteolin, quercetin and their corresponding glycosides also exhibited strong antioxidative activity, while acacetin glucuronide and apigenin-6, 8-di-C-glucoside were relatively less active. Finally, changes in phenolic compositions were also determined by HPLC in the safflower seed and leaf during growth stages and roasting process to produce standardized supplement powerds. These results suggest that phenolic compounds in the roasted safflower seed and leaf may be useful as potential sources of therapeutic agents against several pathological disorders such as carcinogenesis, atherosclerosis and osteoporosis.

• Proceedings of the KSAR Conference
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• 2005.06a
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• pp.47-54
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• 2005

In boars, unlike the cases in males of other species, gonadal hormones suppress voluntary feed intake. for this reason, barrows, compared with gilts or boars, eat too much feed resulting in excessive fat deposition. Two experiments were performed in the present study to investigate the effects of implantation of Revalor H[Experiment(Exp.) I: 140mg trenbolone acetate(a synthetic androgen) + 14mg estradiol-$17\beta(E_{2}\beta)$] and Compudose(Exp. II; 24mg $E_{2}\beta$) on growth efficiency, carcass characteristics and circulating concentrations of IGF-I and IGF-binding protein-3(IGFBP-3). In Exp. I, sixty-four cross-bred finishing barrows weighing approximately 60kg were randomly divided into eight pens under a 2[control vs Revalor implant] $\times$ 2(ad libitum vs $80\%$ ad libitum feeding) $\times$2[control($103\%$ NRC-recommended level) vs low-energy($87\%$ NRC recommendation) diet] arrangement of treatments. In Exp. II, effects of Compudose were studied using 80 finishing barrows(10 animals/pen). In both Exps., all the animals were slaughtered at 100- to 110-kg body weight. Both Revalor and Compudose implants caused a decrease in feed intake and backfat thickness without affecting major physicochemical characteristics of the carcass and an increase in circulating IGF-I concentration. Moreover, Revalor implant exhibited greater effects than restricted feeding, low-energy diet, or Compudose in these variables. In addition, Revalor implantation suppressed weight gain, but enhanced the feed efficiency without exhibiting any interaction with the diet or feeding. In summary, results suggest that 1) both androgen and estrogen suppress voluntary feed intake and backfat deposition and enhance IGF-I secretion and 2) these effects of the gonadal steroid hormones in growth are likely to be mediated, in part, by IGF-I in finishing barrows.

• Journal of Fisheries and Marine Sciences Education
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• v.26 no.5
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• pp.935-940
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• 2014

To investigate effect of on sexual maturation in spotted seabass, Lateolabrax maculatus the sexual maturation, photoperiods were established to natural photoperiod, 9L:15D, 11L:13D, 13L:11D and 15L:9D. After rearing for 90 days from August 11 to November 10, gonadosomatic index (GSI) and hormones were analyzed. In the female, GSI was $1.07{\pm}0.09%$ in the natural photoperiod, $1.93{\pm}0.06%$ in 9L:15D, while the GSI of male was $0.75{\pm}0.05%$ in the natural photoperiod, $1.89{\pm}0.12%$ in 9L:15D and $0.22{\pm}0.05%$ in 15L:9D and the lowest in 15L:9D, respectively. The blood estradiol (E2) of female was $226.0{\pm}13.6pg/ml$ in the natural photoperiod, and the highest value of $296.3{\pm}15.0pg/ml$ in 9L:15D. The male $17{\alpha}20{\beta}OHP$ was similar to the female with $196.3{\pm}17.0pg/ml$ in the natural photo period and $133.0{\pm}15.5pg/ml$ in 9L:15D. The testosterone (T) of male was $259.3{\pm}33.1pg/ml$ in the natural photoperiod, but the hightest value of $370.5{\pm}27.5pg/ml$ in 9L:15D. These results provide basic information from sexual maturation of spotted seabass.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.2
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• pp.282-290
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• 1997

Hepatocytes of Anguilla japonica have been prepared using a collagenase perfusion technique. The isolated cells attached efficiently to fibronectin-coated culture dishes and subsequently formed monolayers in serum-free medium. These cultures maintained in appropriate medium at least for 10 days with minimal cell loss. The effects of estradiol and pituitary hormones on vitellogenin (Vg) synthesis were examined in primary hepatocyte culture of the immature eels. In fish, as in other oviparous vertebrates, estrogen is a major inducer of Vg synthesis. However, $estradiol-17\beta(E_2)$ alone was insufficient to induce Vg synthesis in cultures of eel hepatocytes. Combination of $E_2$ with growth hormone (GH) and/or prolactin (PRL) markedly stimulated Vg synthesis. Even in cultures exposed to $E_2$ or precultured without hormones for 8 days, $E_2$ alone could not fully induce Vg synthesis. The synthesis of Vg was dramatically increased when hepatocytes were cultured in medium supplemented with $E_{2}+GH+PRL$ for 6 days. At this point, even though GH and/or PRL were eliminated from the medium, Vg synthesis was not influenced by these factors during culture of further 3 days. These results indicate that pituitary hormones, in particular GH and PRL, play important roles in the regulation of Vg synthesis in primary cultures of eel hepatocytes.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.149-156
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• 1999

Steroid hormone is known to cause the dynamic changes of mammalian uterus during reproductive cycle, which are modulated via hypothalamus-pituitary -gonad reproductive endocrine axis. Although there were so many studies about estrogenic regulation of uterine growth and differentiation. There is little information about the effect of estrogen on the expression of various transcription factors involved in gene expression. Thus the present study was designed to demonstrate E induced expression of c-fos, c-jun, hsp25 mRNA in rat uterus. Employing Northern blot analysis, we studied the temporal expressions of c-fos, c-jun, and hsp25 messenger RNAs (mRNAs) elicited by a single 17beta-estradiol (E) treatment in the uteri of bilaterally ovariectomized adult rats. c-fos, c-jun, and hsp25 mRNA levels were increased and peaked at 3h after E administration, and then c-fos and c-jun mRNA levels were rapidly decreased to basal control level while, increased hsp25 mRNA levels were sustained till 12h post E treatment. To test the estrogenic effect on the increase of c-fos, c-jun, and hsp25 mRNA levels, we also examined the effects of antiestrogen (tamoxifen). Pretreatment with tamoxifen effectively blocked the E-induced increase of c-fos, c-jun, and hsp25 mRNA levels at 3h post E treatment. Present results suggest that transient increase of c-fos and c-jun protooncogene mRNA at the early time and simultaneous expression of hsp25 mRNA contribute to the response of uterine tissues to E in adult female rats.

• Korean Journal of Animal Reproduction
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• v.19 no.3
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• pp.217-226
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• 1995

The objective of this experiment was to test the effect of EG F on nuclear maturation of pig immature oocytes in vitro. Basic medium used TCM-199 supplemented with 0.2 mM pyruvate, 1 ${\mu}\textrm{g}$/ml estradiol-I7$\beta$ and 25 ${\mu}\textrm{g}$/ml gentamycin, this medium treated with EGF, FSH and FBS. Experiment 1 examined to the effect according to the addition of FSH or EGF (0, 1. 10 and 100 ng EGF/ml) in oocytes maturation. Nuclear maturation rates (M ll %) of 1, 10 and 100 ng EGF/ml (83.0. 8fi.7 and H7.5%) treatments were significantly higher than those of non- and FSH-treated groups (27.3 and 60.3%, p < 0. 001). Experiment 2 examined to the interactive effects of EGF. FSH or FBS during oocytes maturation. Nuclear maturation rates (M ll %) of EGF alone, EGF plus FSH, EGF plus FBS, FSH plus FBS, and EGF plus FSH added FBS treatments (86.7, 90.2, 87.1. 89.6% and 92.6%) were significantly higher than those of non, FSH, and FBS alone treatments (22.3, 52.2 and 42.3%, p < 0.001). Also, cumulus cells expansion of oocytes maturation was examined to total treatments. Normal cumulus cells expansion was shown by FSH plus FBS, EGF or EGF with FBS combination treatments, but cumulus cells of oocyte complexes were still clumped together in EGF-treated groups although they had separated from oocytes. However, EGF showed a positive on nuclear maturation. These results conclude that EGF alone can stimulate nuclear maturation in pig immature oocytes.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.6
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• pp.614-620
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• 2002

The sea bass, Lateoiabrax japonicus is a highly valued fish in aquaculture in South Korea. For establishment of seedling production of sea bass,1 japonicus, we examined change of gonadal development and plasma steroid levels of sea bass reared in indoor tank. Male matured unsimultaneously faster than females and spawning of females took place between the end of January and March. After the spawning period, and until the following January, all the females were in preyitello genesis and in some males, spermatogenetic activity restarted gradually. In October, under reducing photoperiod, cortical alveoli appeared in growing oocyte and the development of spermatogenesis greatly increased. Between October and february, vitellogenesis and spermatogenesis occurred respectively in female and male and gonadosomatic index increased from 4.31 to $24.07\%$ in female and upper 6o/o in male. Also, two sex hormones were analyzed during the course of a reproductive cycle in the sea bass: plasma levels of the gonadal steroid testosterone (T) and estradiol-l7$\beta$ (E_{2}). Variation of the plasma concentrations of T and E, appeared to depend on gonad stages. Plasma T and E, levels were high from November to January, suggesting that an sufficient gonadal stimulation by both hormones may undergoing a processes for the formation of sperm and oocyte.

• Fisheries and Aquatic Sciences
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• v.4 no.4
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• pp.180-185
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• 2001

Al and Cd-induced inhibition of vitellogenin (VTG) production was examined at the estrogen receptor (ER) level in rainbow trout Oncorhynchus mykiss hepatocytes. The binding of $[^3H]$ $estradiol-17\beta\;(E_2)$ to hepatocytes reached a plateau 3 days after addition of $E_2\;(2\times\;10^{-6} M)$to the medium. The binding activity was linearly reduced with the increased concentrations $(-10^{-5}\;M)$ of 4-hydroxy-tamoxifen (4-OHT) and specific binding linearly increased with the increased doses of $[^3H]\;E_2$, indicating that the radioligand bound to ER. Al $(-10^{-4}\;M)$and Cd $(10^{-6}\;M)$ as well as 4-OHT $(10^{-6}\;M)$ significantly reduced the $[^3H]\;E_2$-binding activity by $30­40\%$, while they completely inhibited VTG production. Al and Cd had no effect on $E_2-human$ $ER\alpha$ binding activity at any concentrations used $(-10^5\;nM\;each)$. These results suggested that Al and Cd inhibited VTG production in part by interfereing with the ER level. Inhibitory effects of these metals on the $E_z-dependent$ upregulation of ER activity are also discussed.

• Fisheries and Aquatic Sciences
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• v.9 no.2
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• pp.64-69
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• 2006

Cytochrome P450 (CYP1A) gene expression in the liver and sex steroid levels in plasma were investigated in olive flounder (Paralichthys olivaceus) exposed to tributyltin (TBT) and benzo[a]pyrene (BaP). We constructed a cDNA library and cloned a 230-base sequence encoding partial CYP1A DNA. The CYP1A gene expression level was estimated using northern blotting. Hepatic CYP1A mRNA levels in fish injected with BaP at 10 mg/kg body weight (b.w.) increased for 48 h after injection. However, fish injected with both BaP and TBT at 10 mg/kg b.w. showed no significant changes in CYP1A mRNA level after 48 h. Plasma concentrations of testosterone and $17{\beta}$-estradiol were not significantly different in males and females injected with BaP and TBT. We suggest that TBT-induced suppression of BaP bioactivity should be interpreted with caution in biomonitoring field studies.

• Toxicological Research
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• v.16 no.2
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• pp.141-146
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• 2000

Phthalate esters are used extensively as a plasticizer in the manufacture of plastic products such as PVC bags and medical devices. Recently, phthalate esters have been shown to induce endocrine system mediated responses. However. only a Jew studies have been conducted for estrogenic activity of phthalate esters. In this study estrogenic activities of seven phthalate esters. butyl benzyl phthalate (BBP), di(2-ethylhexyl) phthalate (DEHP), di-n-butylphthalate (DBP), diethylphthalate (DEP), di-n-pentylphthalate (DPP), di-n-propylphthalate (DPrP) and dicyclohexylphthalate (DCHP), were examined in vitro using E-screen assay and competitive binding assay. From the E-screen assay, BBP. DEHP. DBP and DEP showed weak estrogenic activity at the concentration of 5 $\mu\textrm{M}$. The relative proliferative effect (RPE) and the relative proliferative potency (RPP) were 50~70% and 0.01%. respectively, when compared with 500 pM of 17$\beta$-estradiol (E2). In competitive binding assay with the rat uterine estrogen receptor (ER), BBP and DEP showed weak binding potency ［(l/$10^4$~1/$10^5$ of E2］ while DEHP and DBP scarcely bound to ER. These results suggest that some phthalate esters have weak estrogenic activities in vitro.