• The Korean Journal of Physiology and Pharmacology
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• 제17권3호
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• pp.203-208
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• 2013

As the abnormal proliferation of vascular smooth muscle cells (VSMCs) plays a critical role in the development of atherosclerosis and vascular restenosis, a candidate drug with antiproliferative properties is needed. We investigated the antiproliferative action and underlying mechanism of a newly synthesized naphthoquinone derivative, 5,8-dimethoxy-2-nonylamino-naphthalene-1,4-dione (2-nonylamino-DMNQ), using VSMCs treated with platelet-derived growth factor (PDGF). 2-Nonylamino-DMNQ inhibited proliferation and cell number of VSMCs induced by PDGF, but not epidermal growth factor (EGF), in a concentration-dependent manner without any cytotoxicity. This derivative suppressed PDGF-induced $[^3H]$-thymidine incorporation, cell cycle progression from $G_0/G_1$ to S phase, and the phosphorylation of phosphor-retinoblastoma protein (pRb) as well as the expression of cyclin E/D, cyclin-dependent kinase (CDK) 2/4, and proliferating cell nuclear antigen (PCNA). Importantly, 2-nonylamino-DMNQ inhibited the phosphorylation of PDGF receptor${\beta}$(PDGF-$R{\beta}$) enhanced by PDGF at $Tyr^{579}$, $Tyr^{716}$, $Tyr^{751}$, and $Tyr^{1021}$ residues. Subsequently, 2-nonylamino-DMNQ inhibited PDGF-induced phosphorylation of STAT3, ERK1/2, Akt, and $PLC{\gamma}1$. Therefore, our results indicate that 2-nonylamino-DMNQ inhibits PDGF-induced VSMC proliferation by blocking PDGF-$R{\beta}$ autophosphorylation, and subsequently PDGF-$R{\beta}$-mediated downstream signaling pathways.

• 한국수정란이식학회지
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• 제30권2호
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• pp.91-97
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• 2015

Amphiregulin (AREG), a glycoprotein that is a member of the epidermal growth factor (EGF) family, is expressed by the porcine conceptus and endometrium. AREG genotypes were determined based on an SNP in the intron 3 of the gene. Contradictory effects of AREG genotypes on reproductive traits in different pig breeds were reported previously. G allele had undesirable effect on reproductive trait in Meishan breed, while it had favorable effects in Polish Landrace and Large White. We determined AREG genotypes of 179 pigs including the Duroc, Landrace, Yorkshire, Korean native pig (KNP), and Meishan breeds. Two new SNPs were identified near the previously reported SNP in the intron 3 of AREG. Frequencies of AREG alleles among the Duroc, Landrace, Yorkshire, and KNP sows were significantly different (p<0.001), indicating association between AREG genotypes and pig breeds. The first parity litter size was significantly affected by the breeds (p=0.014), but not by AREG genotypes (p=0.148). However, there were breed and AREG genotype associated trends in the first parity litter size. The first parity litter size appeared to be higher in Duroc and KNP sows with G allele, while it appeared to be lower in Landrace sows with G allele. Significant variability of AREG alleles among pig breeds, for the first time in Duroc and KNP sows, was identified. AREG genotypes may influence reproductive traits differentially for each breed and thus, AREG genotypes may need to be considered when sows are bred to increase litter size.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.131-136
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• 1994

In the present study, in order to make an in vitro model of gastric mucosa for physiological and pharmacological studies, we established two immortalized gastric surface mucous cell lines (GSM06 and GSM10), which produce periodic acid-Schiff (PAS)-and concanavalin A (Con A)-positive glycoproteins, from a primary culture of gastric fundic mucosal cells of adult transgenic mice harboring a temperature-sensitive simian virus 40 large T-antigen gene 〔1]. Gastric fundic mucosal cells were isolated as a modification of a previously described method for rats by Schepp et al. (2). The isolated gastric fundic mucosal cells were cultured in DME/F12 medium supplemented with 2% fetal bovine serum (FBS), 1% ITES (consisting of 2 mg/1 insulin, 2 mgg/1 transferrin, 0.122 mg/1 ethanolamine and 0.00914 mg/1 sodium selenite) and 10 ng/ml recombinant epidermal growth factor (EGF) in a collagen-coated culture dish. To remove fibroblastic cells from the culture, gastric mucosal cells were incubated in the culture medium containing dispase (25 U/ml) for 24 h. The cells, uncontaminated with fibroblastic cells, were then cloned by colony formation. In our series of three attempts, two cell lines (GSM06 and GSM10) have been established at last. The cells proliferated, attached to the dish ana grew until confluent monolayers were formed, and maintained tight contact with neighboring cells. Both GSM06 and GSM10 cells have now been in culture for more than 9 months with regular passaging. The either cell produced

• 대한의생명과학회지
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• 제12권1호
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• pp.9-16
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• 2006

We investigated the effects of 835-MHz electromagnetic field (EMF), one of the most popular communication frequency band in Korean code-division multiple-access (CDMA) mobile phone system, on cellular signal transduction. For this, we examined the change of intracellular calcium $([Ca^{2+}]_i)$, reactive oxygen species (ROS) and F-actin polymerization after exposure to 835-MHz EMF followed by the treatment of agonists in Rat-2 fibroblast cells. Culture cells were pretreated with serum-tree medium and concomitantly exposed to 835-MHz at specific absorption rate (SAR) of 4.0 W/kg for 24 hr in a specialized designed apparatus based on Transverse Electro Magnetics (TEM) wave theory. Intracellular $Ca^{2+}$ responses to lysophosphatidic acid (LPA) and epidermal growth factor (EGF) in Rat-2 fibroblast after exposure to 835-MHz EMF were shown to be similar pattern as observed in normal cultured cells. However, the LPA-induced calcium spiking was slightly delayed to 7 sec and sustained thereafter to a little higher ground level under 835-MHz EMF radiation compared to unexposed cells. ROS production level by LPA in the exposed cells was not different from that in control. Furthermore, LPA induced the production of stress fibers with no significant difference in the exposed and unexposed cells. These results suggest that mobile phone radiation (835-MHz, SAR 4.0 W/kg) may not be directly related to signal transduction in Rat-2 fibroblasts except the slight effect of calcium spiking in LPA-induced cells but remain to be further elucidated for possible indirect intervention.

• 한국수정란이식학회지
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• 제15권3호
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• pp.271-277
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• 2000

This study was conducted to establish the optimal culture conditions for in vitro production of bovine embryos derived from slaughter house ovaries. Cumulus-oocyte- complexes (COCs) collected by aspiration from follicles of 2~7 mm in diameter were matured in Ham's F-10 medium supplemented with 0.01 $\mu\textrm{g}$/m1 epidermal growth factor (EGF) at 39$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. After 24 hrs of culture, the oocytes were co-cultured with epididymal sperm selected off by Percoll-density gradient in TALP medium for 24 hrs. The presumptive zygotes were cultured in HECM-6 medium for 3 d post-insemination, and followed by cultured in TCM199 medium until 7 to 10d post-insemination. The cultures were compared of their cleavage and development into later stage in culture medium by additions of different protein sources (PVA, BSA and BCS) and by different embryo density. The rates of cleavage and development rates into blastocyst were not significantly (P<0.05) different among the culture media containing with BSA (75.0% and 40.5%), BCS (76.7% and 38.0%) and PVA (72.5% and 42.2%), respectively. Significantly (P<0.05) higher blastocysts rates were obtained in culturing of 30 and 40 embryos in each 50$\mu$l droplets of culture medium than in 5, 10 and 20 embryos. These results indicate that the optimal density of embryos is 30~40 embryos in a 50$\mu$l droplet of culture medium. Furthermore there is no effect of different protein sources on early embryonic development.

• Asian-Australasian Journal of Animal Sciences
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• 제30권1호
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• pp.20-33
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• 2017

Objective: Superstimulatory treatment of one-month-old lambs can achieve synchronous development of numerous growing follicles. However, these growing follicles cannot complete maturation and ovulation. Oocyte maturation and competence are acquired during follicular development, in which granulosa cells play an essential role. Methods: In this study, we applied RNA sequencing to analyze and compare gene expression between prepubertal and adult superstimulated follicle granulosa cells in sheep. Results: There were more than 300 genes that significantly differed in expression. Among these differently expressed genes, many extracellular matrix genes (EGF containing Fibulin Like Extracellular Matrix Protein 1, pentraxin 3, adrenomedullin, and osteopontin) were significantly down-regulated in the superstimulated follicles. Ingenuity pathway and gene ontology analyses revealed that processes of axonal guidance, cell proliferation and DNA replication were expressed at higher levels in the prepubertal follicles. Epidermal growth factor, T-Box protein 2 and beta-estradiol upstream regulator were predicted to be active in prepubertal follicles. By comparison, tumor protein P53 and let-7 were most active in adult follicles. Conclusion: These results may contribute to a better understanding of the mechanisms governing the development of granulosa cells in the growing follicle in prepubertal sheep.

• Asian Pacific Journal of Cancer Prevention
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• 제15권14호
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• pp.5889-5893
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• 2014

Over-expression of epidermal growth factor receptor (EGFR) has been identified as a common feature associated with clinical outcome in many types of cancer, including squamous cell carcinoma of the oesophagus (SCCO). However, the clinical importance of EGFR over-expression in SCCO remains unsettled as conflicting results exist. Therefore we carried out the present meta-analysis of published studies for clarification. A total of 13 studies including 1, 150 patients were enrolled. EGFR over-expression was positive in 722 of these cases. With EGFR over-expression, patients had higher depth of invasion, vascular invasion, and poor prognosis. However, expression had no relation with degree of differentiation, histological grade, lymph node metastasis, clinical stage or lymphatic invasion. EGFR over-expression is probably a valuable predictor for the T stage, vascular invasion and OS, and it could be used as a poor prognosis indicator for the esophageal SCC patients. Targeting therapy to EFGR should be considered to the combined treatment in SCCO.

• 문화기술의 융합
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• 제4권3호
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• pp.197-207
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• 2018

무의 껍질과 무청은 식용으로 사용하기도 하지만, 질기고 거친 식감으로 인하여 제거 후 부산물로서 폐기되거나 가축의 사료로 이용되는 실정이다. 본 연구는 초임계 열처리가공 무추출물이 UVB로 유도된 Hos:HRM-2 주름생쥐동물모델에서 항-광노화 주름개선 효능을 검증하고자 하였다. 초임계 열처리된 무 추출물을 7주령의 HRM-2 생쥐에 UVB 램프로 등피부에 조사하여 피부 노화를 유발하여 피부두께, 탄력, 그리고 주름생성 변화에 대하여 알아보고자 하였다. 초임계 열처리된 무추출물의 UVB 조사에 의한 HRM생쥐의 주름 측정 결과는 정상군에 비해 음성대조군의 주름의 깊이가 증가되는 것을 관측할 수 있었다. 반면 무 추출물의 처리군에서는 음성대조군에 비해 주름의 깊이가 감소되었다. 초임계 열처리된 무추출물의 UVB 조사에 의한 의한 주름관련 유전자 및 단백질의 발현에 미치는 영향에서는 음성대조군은 정상군에 비해 MMP-2 및 MMP-9 유전자의 발현이 현저하게 증가하였으나, 무 추출물의 처리에 의해 상기 유전자의 발현이 유의적으로 감소하는 것을 확인할 수 있었다. 또한 유전자의 발현 정도는 무 추출물의 처리량에 의존적으로 감소하였다. 실험결과를 종합적으로 해석해 볼 때, 초임계 열처리된 무 추출물은 피부에 도포 시 뿐만 아니라 경구로 섭취시에도 피부 주름 개선에 효과가 있어, 건강기능식품 조성물로도 유용하게 사용될 수 있다. 특히 무는 오랫동안 복용 시에도 전혀 부작용을 나타내지 않는 안전한 식품으로 장기간 꾸준히 복용하는 것에 의해 피부 주름 및 탄력 개선 효과를 얻을 수 있다는 결론을 얻을수 있었다.

• Applied Microscopy
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• 제29권2호
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• pp.163-175
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• 1999

본 실험은 화상을 입은 생쥐에서 SQ의 치유 효과를 알아보기 위함에 있다. 실험군은 7개군으로 구분하였다; 대조군, 화상군, 그리고 화상을 입헌 후 SQ를 처치한 군. 대조군을 제외한 모든 군은 등 부위에 2도 화상을 입혔다. SQ를 처치한 모든 군은 하루에 한 번씩 10초 간격으로 순수한 SQ 3방울을 도포하였다. 화상 후 10일 동안 조직학적 및 미세구조적 변화를 광학 현미경과 전자현미경으로 관찰하였다. 광학현미경으로 관찰한 결과, 화상군의 표피층의 모든 부분은 SQ를 처치한 군보다 더 심한 상처를 입었다. SQ를 처치한 6째와 10일째 군에서, 기저층은 분화가 활발하였고, 가시층의 각화세포는 수적으로 매우 증가하였다. 전자현미경으로 관찰한 결과, SQ를 처치한 모든 군의 기저층에서 세포분열이 화상군에서보다 더 활발하게 나타났다. 특히, SQ를 처치한 10일군에서는 이상적인 결과를 얻을 수 있었다; 대조군보다 더 두꺼운 가시층과 많은 각화세포, 뚜렷한 세포간교, 그리고 멜라닌세포를 포함하는 튼튼한 기저층. 이와 같은 결과로 보아 SQ은 표피성장인자(EGF)를 활성화시키고, 유해산소의 제거 역할을 하며, 막계에 에너지원을 제공할 가능성이 있다. 이 실험의 결과는 SQ가 화상 치료에 특수한 효과가 있다고 사료된다.

• 한국수정란이식학회지
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• 제11권2호
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• pp.125-135
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• 1996

The present study was carried out to investigate the effects of cryoprotectants, equilibration step, freezing rate, culture condition following in vitro fertilization, and age and development stage of embryo by freezing with conventional slow freezing and vitrification on survival of frozen-thawed Korean native cattle(KNC) blastocysts produced in vitro. The KNC blastocysts produced in vitro were equilibrated in 1.8M ethylene glycol or 1.4M glycerol and cooled from -6$^{\circ}C$ to -35$^{\circ}C$ at -0.3$^{\circ}C$ or -O.6$^{\circ}C$ /minute. When equilibrated in 1.8M ethylene glycol, survival rate of fiozen4hawed blastocysts was sarne in both -0. 3$^{\circ}C$ /min and -0.6$^{\circ}C$ /min cooling rate(71.4%). With the equilibration in 1.4M glycerol, survival rate was higher in -0.3$^{\circ}C$ /min(63.6%) than in -0.6$^{\circ}C$ /min cooling rate(53.8%). For vitrification of the KNC blastocysts produced in vitro, they were equilibrated in 2-step or 3-step exposure to vitrification solution(25% ethylene glycol + 25% glycerol). Survival rate was sirilar in both 2-step(45.0%) and 3-step exposure(47.4%). According to culture condition following in vitro fertilization, higher survival rate was obtained for blastocysts co-cultured with bovine oviductal epithelial cell(BOEC, 77.3%) than for those cultured with epidermal growth factor(EGF, 65.7%) or for those co-cultured with BOEG + EGF (54.8%). According to embryo age and development stage, higher survival rate was obtained for 7-day ernbryos(70.0%) than 8-day(56.8%) or 9-day(20.0%) for blastocyst stage and obtained for 8-day embryos(74.3%) than 7-day(62.5%) or 9-day(42.9%) for exponded blastocyst. In surnmary, higher survival rate of frozen4hawed KNC blastocysts produced in vitro were obtained by using ethylene glycol for cryoprotectant and -0.3$^{\circ}C$ /min for cooling rate. And higher survival rate were obtained with co-culture with BOEC for culture condition following in vitro fertilization and with 7-day blastocyst or 8-day expanded blasto cyst for embryo age and development stage.