• Korean Journal of Animal Reproduction
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• v.19 no.2
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• pp.95-104
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• 1995

Mouse mammary epithelial cells(NMuMG) were maintained onto 6-well plates (3$\times$105 cells/well) or chambered slide (1$\times$104 cells/well), in DMEM supplemented with 10% fetal calf serum. After serum starvation for 24 hours, DMNB (1$\mu$M) was added and exposed to UV light (300nm, 3 second pulse) after 2 hours from DMNB addition in order to activate DMNB which induces a rapid transient increase in intracellular cAMP upon UV irradiation. EGF (100ng/ml) and/or IGF-I (10ng/ml) were treated at the time of UV irradiation. Nuclear labeling index was estimated as percent of nuclear labeled cells(percent of S phase of cells) by incorporation of 3H-thymidine into DNA(1 hour pulse with 1$\mu$Ci/ml). DMNB(1$\mu$M), EGF (100ng/ml) and/or IGF-I (10ng/ml) signifciantly increased nuclear labeling index than those of control (P<0.05). Addition of DMNB＋EGF or DMNB＋EGF＋IGF-I showed the interaction effect in nuclear labeling index (P<0.05). Protein kinase A activities by addition of EGF, IGF-I or EGF＋IGF-I were 10.5, 9.8 or 9.4 unit/mg protein, respectively, and no statistical difference was found in comparison with control (P>0.05). Additon of DMNB＋EGF showed the moderate interaction effect on tyrosyl kinase activity (P<0.1). In the fluorography analysis, there were no specific protein phosphorylation patterns were found at 1 or 15 minute by addition of DMNB. EGF and/or IGF-I. These results suggest that the interaction effect in nuclear labeling index by addition DMNB and EGF could be mediated through the modulation of tyrosyl kinase activity by cAMP.

• The Korean Journal of Zoology
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• v.39 no.2
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• pp.215-222
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• 1996

In the previous studies, we have demonstrated that prorenin converting enzyme (PRECE) was identical to the epidermal grouch factor-binding protein (EGF-BP) type B, which was a member of the mouse glandular kallikrein family, To examine whether or not EGF-BP type A and C are involved in the processing of prorenin, we have cloned the CDNAS of the EGF-BP type h and C from a library of male ICR mouse submandibular gland (SMGI. And then CHO cells were transfected with the EGF-BP expression plasmids. and stable cell lines expressing a high level of the EGF-BPS precursor were obtained. The conditioned medium was then treated with trypsin, which has been knotvn to effectively convert the EGF-BP type A and C precursor to the active forms. 수ubsequentlv, the prorenin converting activity of the trypsin-treated or untreated medium was examined. PRECE converted exactly prorenin to renin, but the prorenin converting activities of EGF-BP type A and C were not detected. From these results, it seems that only type B of these EGF-BPs is involved in processing Ren 2 prorenin in mouse SMG.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.4
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• pp.493-501
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• 1998

The most significant direct role of estrogen in vivo is its ability to elicit receptor-mediated cellular proliferation in mammalian target tissues. However, the mechanism by which exogenously added estrogen causes the neoplastic transformation of renal cortical cells is yet to be uncovered. The present study was designed to evaluate interaction of $17{\beta}-estradiol\;(E_2)$ with epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) on proliferation and $P_i$ uptake in primary cultured rabbit renal proximal tubular cells in phenol red-free, hormonally defined-medium. $[^3H]-thymidine$ incorporation increased markedly by about 133% and 141% more in the presence of $10^{-9}\;and\;10^{-6}\;M\;E_2$, respectively, than that of control. Cell count was 162% and 143% greater in the presence of $10^{-9}\;and\;10^{-6}\;M\;E_2$ , respectively, compared with control. Among all time points examined, there was an increase in $[^3H]-thymidine$ incorporation in the presence of $10^{-9}\;M\;E_2$ at day 9 or 13, respectively. However, $E_2$ ($10^{-9}\;M$) significantly drove up cell count to 160% of that of control at day 13, while it had a slight but statistically insignificant effect at day 9. $E_2-induced$ stimulation of $[^3H]-thymidine$ incorporation was completely reversed by $E_2$ antagonists (progesterone or tamoxifen). $E_2$ ($10^{-9}\;M$) or EGF ($10^{-8}\;M$) significantly stimulated $[^3H]-thymidine$ incorporation by 144% and 154% of control. $E_2$ plus EGF was synergistic on $[^3H]-thymidine$ incorporation (204% of control), while $E_2$ plus IGF-I showed a slight but no significant synergistic effect. Cell number also displayed similar pattern. $E_2$ ($10^{-9}\;M$) significantly stimulated $P_i$ uptake to 134% of control. $E_2$-induced stimulation of $P_i$ uptake was partially reversed by $E_2$ antagonists. EGF or IGF-I ($10^{-8}\;M$) significantly also increased $P_i$ uptake to 132% or 129% of control. $E_2$ plus EGF had synergistic effect on $P_i$ uptake, while $E_2$ plus IGF-I did not. In conclusion, $E_2$ may act not only directly interaction with its receptors but also indirectly as a modulator of EGF in proliferation and $P_i$ uptake of primary cultured rabbit renal proximal tubular cells.

• Korean Journal of Animal Reproduction
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• v.27 no.2
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• pp.169-177
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• 2003

This study was conducted to determine effects of polyvinyl alcohol (PVA), fetal bovine serum (FBS) bovine serum albumin (BSA) and epidermal growth factor (EGF) on blastocoel formation, total cell number, apoptosis and apoptosis-related gene expression of porcine diploid parthenotes developing in vitro. The addition of 0.4% BSA to the culture medium enhanced the development of 2-cell stage parthenotes to the blastocysts stage (P<0.01). FBS reduced cell numbers of blastocysts (P<0.01) and increased percentage of apoptosis in the blastocysts (P<0.001). However, while BSA increased cell numbers, it did so only when EGF was present. Either agent on its own had no effect. Similarly, apoptosis in the blastocysts was not influenced by either agent on its own but was reduced when both BSA and EGF were present. Furthermore, semi-quantitative reverse-transcriptase polymerase chain reaction revealed that EGF enhanced the mRNA expression of Bcl-xL in the presence of 0.4% BSA but BSA and EGF alone had no effect, and EGF and/or BSA did not influence Bak gene expression in the blastocyst stage parthenotes. However FBS reduced Bcl-xL mRNA expression (P <0.05) and enhanced Bak expression. This result suggests that apoptosis related genes expression is significantly affected by supplements, which may result in alteration of apoptosis and embryo viability of porcine embryos developing in vitro.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.8
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• pp.1117-1123
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• 2009

This study investigated the effects of IGF-I and EGF on the development of blastocysts or hatched blastocysts during the in vitro culture of embryos from immature porcine oocytes. After the in vitro maturation and fertilization of cumulus-oocyte complexes (COCs) and their culture in vitro in PZM3 medium, we examined the embryo development rate for 168 h. When different concentrations of IGF-I (0, 1, 10, 20 ng/ml) were supplemented to fertilized porcine embryos in vitro, there were no significant differences in cleavage rate, blastocyst development rate or blastocyst hatching rate among the treated groups. On the other hand, when different concentrations of EGF (0, 1, 10, 20 ng/ml) were supplemented to the in vitro culture medium, blastocyst development rate was highest in the group in which EGF was not supplemented and, specifically, it was higher than in the 20 ng/ml treatment group (p<0.05). When 10 ng/ml IGF-I and 1 ng/ml EGF were supplemented separately or simultaneously, there were no significant differences among the treated groups in blastocyst hatching rate and the number of cells in each condition. This study demonstrated that the addition of IGF-I and EGF into PZM3 medium did not enhance development of the blastocyst stage and total cell number in blastocysts.

• Natural Product Sciences
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• v.23 no.3
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• pp.201-207
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• 2017

Angelica decursiva has been utilised as remedy for controlling the airway inflammatory diseases in folk medicine. We investigated whether nodakenin, columbianadin, and umbelliferone isolated from the roots of Angelica decursiva inhibit the gene expression and production of MUC5AC mucin from human airway epithelial cells. Confluent NCI-H292 cells were pretreated with nodakenin, columbianadin or umbelliferone for 30 min and then stimulated with epidermal growth factor (EGF), phorbol 12-myristate 13-acetate (PMA) or tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) for 24 h. The MUC5AC mucin gene expression was measured by reverse transcription - polymerase chain reaction (RT-PCR). Production of MUC5AC mucin protein was measured by enzyme-linked immunosorbent assay (ELISA). The results were as follows: (1) Nodakenin did not affect the expression of MUC5AC mucin gene induced by EGF, PMA or $TNF-{\alpha}$. Columbianadin inhibited the expression of MUC5AC mucin gene induced by EGF or PMA. However, umbelliferone inhibited the expression of MUC5AC mucin gene induced by EGF, PMA or $TNF-{\alpha}$; (2) Nodakenin also did not affect the production of MUC5AC mucin protein induced by EGF, PMA or $TNF-{\alpha}$. Columbianadin inhibited the production of MUC5AC mucin protein induced by PMA. However, umbelliferone inhibited the production of MUC5AC mucin protein induced by EGF, PMA or $TNF-{\alpha}$. These results suggest that, among the three compounds investigated, umbelliferone only inhibits the gene expression and production of MUC5AC mucin stimulated by various inducers, by directly acting on airway epithelial cells, and the results might explain the traditional use of Angelica decursiva as remedy for diverse inflammatory pulmonary diseases.

• Development and Reproduction
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• v.14 no.1
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• pp.27-34
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• 2010

A fertilized oocyte can get the competence for implantation through cleavage and stage-specific gene expression. These are under the control of autonomous and exogenous regulators including physiological culture condition. Endogenous and exogenous growth factors are considered as critical regulators of cleaving embryos during travel the oviduct and uterus. In this study, an effort was made to evaluate comprehensively the quality of embryos for implantation, grown in media enriched with EGF and PAF. The study evaluated developmental rates on given time, blastulation and hatching rates, and adhesion rates. Developmental rates of blastocyst to the hatching stage were significantly high in PAF treated group compared to the control in a dose-dependent manner but not in EGF group. Implantation rates were significantly high both PAF and EGF in a dose-dependent manner. H7, a PKC inhibitor, blocked the process of hatching of the blastocysts but combined treatment of EGF and PAF enhanced the hatching and implantation of blastocsyts. Based on these results it is suggested that EGF and PAF support acquirement of implantation competence at blastocyst stage through a PKC pathway.

• Reproductive and Developmental Biology
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• v.29 no.2
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• pp.127-131
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• 2005

The objective of this study was to examine the effect of EGF on meiotic maturation and pronuclear (PN) formation of porcine oocytes. Prepubertal gilt cumulus-oocyte-complexes (COCs) aspirated from $2\~6mm$ follicles of abbatoir ovaries were matured in TCM199 containing 0.1mg/ml cysteine, $0.5{\mu}/ml$ FSH and LH, and EGF (0, 5, 10, 20, 40 ng/ml) for 22 hr at $39^{\circ}C$ in a humidified atmosphere of $5\%$ $CO_2$ in air. They were then cultured for an additional 22hr without hormones. In Experiment 1, to examine the nuclear maturation at 44hr of culture, the expanded cumulus cells were removed by vortexing for 1 min in 3 mg/ml hyaluronidase. The oocytes were fixed in acetic acid: methanol (1:3, v/v) at least for 48 hr and stained with $1\%$ orcein solution for 5 min. Nuclear status was classified as germinal vesicle (GV), germinal vesicle breakdown (GVBD), prophase-metaphase I (PI-MI), and PII-MII under microscope. In Experiment 2, to investigate PN formation, oocytes were fertilized with Percoll-treated freshly ejaculated sperm $(1\times10^5\; cells/ml)$ in mTBM with $0.3\%$ BSA and 2mM caffeine for 5hr, and cultured in NCSU-23 medium with $0.4\%$ BSA. At 6hr of culture, the embryos were fixed in $3.7\%$ formaldehyde for 48hr and stained with 10ug/ml propidium iodide for 30 min. PN status was classified as no or one PN (unfertilized), 2 PN (normal fertilized) and $\geq3$ PN (polyspermy). Differences between groups were analyzed using one-way ANOVA after arc-sine transformation of the proportional data. The rate of oocytes that had reached to PII-MII were significantly (P<0.05) higher in all groups added EGF than that of non-treated group $(67\%)$, but it did not differ among the all added groups $(86\%,\;85\%,\;79\%\;and\;81\%$, in 5, 10, 20 and 40 ng/ml EGF, respectively). No differences on the incidence of 2PN were observed in all treated groups $(25\%,\;30\%,\;33\%,\;29\%\;and\;29\%$, in 0, 5, 10, 20 and 40 ng/ml EGF, respectively), however, in non-treated group, polyspermy tended to be increased ($66\%\;vs\;. 58\%,\;54\%,\;52\%\;and\;55\%$, 0 vs. 5, 10, 20, 40 ng/ml EGF, respectively). These results suggest that EGF can be effectively used as an additive for enhancing oocyte maturation and reducing the incidence of polyspermy in pig.

• Archives of Pharmacal Research
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• v.28 no.11
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• pp.1311-1316
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• 2005

To better define the relationship between dermal regeneration and wound contraction and scar formation, the effects of epidermal growth factor (EGF) loaded in collagen sponge matrix on the fibroblast cell proliferation rate and the dermal mechanical strength were investigated. Collagen sponges with acid-soluble fraction of pig skin were prepared and incorporated with EGF at 0, 4, and 8 $\mu$g/1.7 $cm^{2}$. Dermal fibroblasts were cultured to 80$\%$ confluence using DMEM, treated with the samples submerged, and the cell viability was estimated using MTT assay. A deep, $2^{nd}$ degree- burn of diameter 1 cm was prepared on the rabbit ear and the tested dressings were applied twice during the 15-day, post burn period. The processes of re-epithelialization and dermal regeneration were investigated until the complete wound closure day and histological analysis was performed with H-E staining. EGF increased the fibroblast cell proliferation rate. The histology showed well developed, weave-like collagen bundles and fibroblasts in EGF-treated wounds while open wounds showed irregular collagen bundles and impaired fibroblast growth. The breaking strength (944.1 $\pm$ 35.6 vs. 411.5 $\pm$ 57.0 Fmax, $gmm^{-2}$) and skin resilience (11.3 $\pm$ 1.4 vs. 6.5 $\pm$ 0.6 mJ/$mm^{2}$) were significantly increased with EGF­treated wounds as compared with open wounds, suggesting that EGF enhanced the dermal matrix formation and improved the wound mechanical strength. In conclusion, EGF-improved dermal matrix formation is related with a lower wound contraction rate. The impaired dermal regeneration observed in the open wounds could contribute to the formation of wound contraction and scar tissue development. An extraneous supply of EGF in the collagen dressing on deep, $2^{nd}$ degree-burns enhanced the dermal matrix formation.

• YAKHAK HOEJI
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• v.40 no.4
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• pp.456-461
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• 1996

The effects of human epidermal growth factor(EGF) which was produced by recombinant DNA technique was investigated on gastric secretion, gastric lesion and ulcer models in rats. The EGF showed significant inhibition of secretion of gastric juice and total acid output, at 0.4mg/kg, id and also inhibited Shay ulceration at 0.4mg/kg, id in rats. The lesion induced by absolute ethanol was significantly reduced by oral administration of EGF at 0.4mg/kg. Likewise, EGF caused significant inhibition of indomethacin induced gastric ulcer at oral doses of 0.2 and 0.4mg/kg. The EGF produced dose-dependent inhibition of gastric ulcer induced by acidified aspirin, but showed no significant inhibition at oral doses of 0.1, 0.2 and 0.4mg/kg. The chronic gastric ulcer induced by injection of 20% acetic acid solution was significantly reduced by oral doses of 0.1 and 0.4mg/kg of EGF. Duodenal ulcer induced by mepirizole was dose-dependently inhibited by oral doses of 0.1, 0.2 and 0.4mg/kg of EGF. These data suggest that EGF possesses pronounced inhibitory action in gastric ulcer and duodenal ulcer of rats.