• Clinical and Experimental Reproductive Medicine
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• 제39권2호
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• pp.73-80
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• 2012

Objective: This study was undertaken to determine the effect of hypoxia inducible factor (HIF)-$1{\alpha}$ on the cell death, autophagy, and invasion of trophoblasts. Methods: To understand the effect of HIF-$1{\alpha}$, we inhibited HIF-$1{\alpha}$ using siRNA under normoxia and hypoxia conditions. Invasion assay and zymography were performed to determine changes in the invasion ability of HIF-$1{\alpha}$. Western blotting and immunofluorescence were performed to determine some of the signal events involved in apoptosis and autophagy. Results: There was no difference in cell death through the inhibition of HIF-$1{\alpha}$ expression by siRNA; however, the expression of LC3 and autophagosome formation increased. On the other hand, autophagy was increased, and the invasive ability of trophoblast cells decreased according to the inhibition of HIF-$1{\alpha}$ expression by siRNA. These experimental results mean that HIF-$1{\alpha}$ genes regulate the invasive ability of trophoblasts by increasing autophagy. Conclusion: This study contributes important data for understanding the mechanism of early pregnancy implantation and the invasive ability of trophoblasts by defining the relationship between the roles of HIF-$1{\alpha}$ and autophagy.

• Clinical and Experimental Reproductive Medicine
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• 제40권1호
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• pp.12-22
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• 2013

Objective: We investigated the norepinephrine transporter (NET) expression in normal and pre-eclamptic placentas and analyzed the invasion activity of trophoblastic cells based on norepinephrine (NE)-NET regulation. Methods: NET and NE expression levels were examined by western blot and enzyme-linked immunosorbent assay, respectively. Trophoblast invasion activity, depending on NE-NET regulation, was determined by NET-small interfering RNA (siRNA) and NET transfection into the human extravillous trophoblast cells with or without NE treatment and invasion rates were analyzed by zymography and an invasion assay. Results: NET mRNA was expressed at a low level in pre-eclamptic placentas compared with normal placentas and NE concentration in maternal plasma increased significantly in pre-eclamptic women compared to normal pregnant women (p<0.05). NET gene upregulation and NE treatment stimulated trophoblast cell invasion up to 2.5-fold (p<0.05) by stimulating matrix metalloproteinase-9 activity via the phosphoinositol-3-kinase/AKT signaling pathway, whereas NET-siRNA with NE treatment reduced invasion rates. Conclusion: NET expression is reduced by inadequate regulation of NE levels during placental development. This suggests that a complementary balance between NET and NE regulates trophoblast cell invasion activities during placental development.

• IMMUNE NETWORK
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• 제13권6호
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• pp.283-288
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• 2013

The pro-inflammatory cytokines tumor necrosis factor-${\alpha}$ (TNF${\alpha}$) and interleukin (IL)-$1{\beta}$ are crucial mediators involved in chronic inflammatory diseases. Inflammatory signal pathways regulate inflammatory cytokine expression-mediated by p38 mitogen activated protein kinase (p38MAPK). Therefore, considerable attention has been given to p38MAPK as a target molecule for the development of a novel anti-inflammatory therapeutics. BIRB 796, one of p38MAPK inhibitor, is a candidate of therapeutic drug for chronic inflammatory diseases. In this study, we investigated the effect of BIRB 796 on inflammatory cytokine productions by lipopolysaccharide (LPS) in different immune cell types. BIRB 796 reduced LPS-mediated IL-8 production in THP-1 cells but not in Raw 264.7 cells. Further analysis of signal molecules by western blot revealed that BIRB 796 sufficiently suppressed LPS-mediated phosphorylation of p38MAPK in both cell types whereas it failed to block inhibitor of kappa B (I-${\kappa}B$) degradation in Raw 264.7 cells. Taken together, these results suggest that the anti-inflammatory function of BIRB 796 depends on cell types.

• Clinical and Experimental Reproductive Medicine
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• 제41권1호
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• pp.1-8
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• 2014

Objective: Estrogen related receptor ${\beta}$ (Esrrb) is a member of the orphan nuclear receptors and may regulate the expression of pluripotencyrelated genes, such as Oct4 and Nanog. Therefore, in the present study, we have developed a method for delivering exogenous ESRRB recombinant protein into embryos by using cell-penetrating peptide (CPP) conjugation and have analyzed their effect on embryonic development. Methods: Mouse oocytes and embryos were obtained from superovulated mice. The expression of Oct4 mRNA and the cell number of inner cell mass (ICM) in the in vitro-derived and in vivo-derived blastocysts were first analyzed by real time-reverse transcription-polymerase chain reaction and differential staining. Then 8-cell embryos were cultured in KSOM media with or without $2{\mu}g/mL$ CPP-ESRRB protein for 24 to 48 hours, followed by checking their integration into embryos during in vitro culture by Western blot and immunocytochemistry. Results: Expression of Oct4 and the cell number of ICM were lower in the in vitro-derived blastocysts than in the in vivo-derived ones (p<0.05). In the blastocysts derived from the CPP-ESRRB-treated group, expression of Oct4 was greater than in the non-treated groups (p<0.05). Although no difference in embryonic development was observed between the treated and non-treated groups, the cell number of ICM was greater in the CPP-ESRRB-treated group. Conclusion: Treatment of CPP-ESRRB during cultivation could increase embryos' expression of Oct4 and the formation rate of the ICM in the blastocyst. Additionally, an exogenous delivery system of CPP-conjugated protein would be a useful tool for improving embryo culture systems.

• 대한예방한의학회지
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• 제12권3호
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• pp.141-155
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• 2008

Traditional, complementary, and alternative medicine(CAM) constitutes a major source of health care in developing countries. Its therapies, products and practices are increasing used in industrialized countries, typically by around 50% of the population. However, controls on the provider of CAM services and supply and promotion of complementary medicines are weak in these countries. These trends evoke important public health questions relating to benefits and detriments, safety and quality from medical, financial, and social perspectives. In the western counties, investment in research is increasing, and some guideline is now available for ensuring their quality, efficacy, and safety according to the criteria of evidence-based medicine as legally demanded. National policies and legislation are being developed in many countries and often include partnerships between biomedical and traditional health practitioners. The purposes of the study were to review the policies on CAM, to examine the responses of the government to cope with use of CAM in industrialized countries. Lastly, some implications and recommendation are suggested.

• BMB Reports
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• 제41권2호
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• pp.170-175
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• 2008

In protein therapy, it is important for exogenous protein to be delivered into the target subcellular localization. To transduce a therapeutic protein into its specific subcellular localization, we synthesized nuclear localization signal (NLS) and membrane translocation sequence signal (MTS) peptides and produced a genetic in-frame SOD fusion protein. The purified SOD fusion proteins were efficiently transduced into mammalian cells with enzymatic activities. Immunofluorescence and Western blot analysis revealed that the SOD fusion proteins successfully transduced into the nucleus and the cytosol in the cells. The viability of cells treated with paraquat was markedly increased by the transduced fusion proteins. Thus, our results suggest that these peptides should be useful for targeting the specific localization of therapeutic proteins in various human diseases.

• Clinical and Experimental Reproductive Medicine
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• 제37권3호
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• pp.217-229
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• 2010

목 적: 영양막세포의 과도한 세포자멸사는 태반의 발달뿐 아니라 산과 질환을 유발하는 요인으로 알려져 있다. X-linked inhibitor of apoptosis (XIAP)은 임신 기간 동안 영양막세포의 세포자멸사와 관련되어 있다고 알려져 있으나, 자간전증을 유발하는 인자인 저산소성과의 관계에 관한 연구는 미흡한 실정이다. 본 연구의 목적은 XIAP가 정상 태반과 자간전증 산모의 태반에서 발현 양상의 차이를 분석하고, 저산소 상태에 노출된 HTR-8/SVneo 영양막세포주에서의 XIAP 기능을 분석하고자 하였다. 연구방법: XIAP 발현을 분석하고자, 정상 태반 (n=15), 중기 자간전증 태반 (n=11), 그리고 말기 자간전증 태반 (n=15) 조직을 수집하여 RT-PCR, 면역조직화학법, 그리고 Western blot 등을 실시하였다. 저산소성 상태에서 XIAP의 기능을 확인하고자 HTR-8/SVneo 영양막세포주에 1% 산소가 공급되는 hypoxia 상태에 노출시킨 뒤 12시간, 24시간 후에 각 세포자멸사 관련 유전자들의 발현을 fluorescence-activated cell sorting (FACS)와 Western blot 분석 등을 실시하였다. 결 과: XIAP는 태반의 합포영양막세포와 포합체결절에서 발현이 관찰되었으며, 정상 태반보다 자간전증 태반에서의 발현이 현저히 감소됨이 관찰되었다 (p<0.05). 또한, 저산소 상태에 노출된 HTR-8/SVneo 영양막세포주에서 감소된 XIAP 발현은 세포질에서 핵으로의 이동에 따라 세포자멸사를 유발하는 단백질들의 발현이 증가됨이 관찰되었다. 결 론: XIAP의 발현은 태반 발달 및 자간전증 태반에서 XIAP 유전자의 발현은 감소되었으며, XIAP의 저하로 인한 caspase-9의 증가가 자간전증 태반에서의 세포자멸사는 더 많이 유도되었음을 확인할 수 있었다. 또한, 저산소 상태에 의해 XIAP의 발현이 감소되었으며, XIAP 단백질의 세포질에서 핵으로의 위치 변화는 영양막세포의 세포자멸사에 중요한 역할을 하는 것이 관찰되었고, 이는 자간전증의 진단에 유용한 마커로써의 활용되기 위한 기본적인 자료로 활용될 수 있을 것으로 판단된다.

• 대한의생명과학회지
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• 제19권3호
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• pp.261-265
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• 2013

The corpus luteum is a transient endocrine gland essential for regulation of the ovarian cycle as well as for establishing and maintaining pregnancy. Prostaglandin $F_{2{\alpha}}$ (PGF) initiates functional and structural regression of the corpus luteum and therefore is an important regulator of the estrous cycle. It is a matter of debate whether the endothelial cells of the bovine corpus luteum express PGFR, the cognate receptor for PGF. Therefore, the aim of this study was to assess the expression of PGFR in bovine endothelial cells. Endothelial cells were isolated from the bovine corpus luteum of the mid-luteal stage using magnetic beads and cultured in vitro. We demonstrate that this isolation procedure generates a pure culture of endothelial cells as confirmed by synthesis of Factor VIII and lack of expression of $3{\beta}$-hydroxysteroid dehydrogenase. By RT-PCR, Western blot and immunofluorescence analyses, we further show that the cultured endothelial cells produced PGFR. This model system can be utilized to provide an experimental system to investigate the role of PGF on endothelial cells during the reproductive cycle.

• Advances in Traditional Medicine
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• 제10권4호
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• pp.304-309
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• 2010

Scutellaria pinnatifida A. Hamilt. (Lamiaceae) is an endemic Turkish herb. This plant is also endemic to Iran, and grows abundantly in other central and western Asian countries. Several species of the Scutellaria are known for their traditional uses in the treatment of hypertension, arteriosclerosis, inflammatory diseases, hepatitis, allergy, cancer and diarrhoea. Free-radical-scavenging property, antibacterial activity and brine shrimp toxicity of the n-hexane, dichloromethane (DCM) and methanol (MeOH) extracts of S. pinnatifida were assessed using the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assay, the resazurin microtitre plate based assay, and the brine shrimp lethality assay, respectively. The DCM and MeOH extracts exhibited free-radical-scavenging property, with the $RC_{50}$ values of 0.362 and 0.127 mg/ml, respectively. Among the solid-phase extraction fractions of the MeOH extract, the 50% aqueous-MeOH fraction showed the highest level of free-radicalscavenging activity ($RC_{50}$ = 0.039 mg/ml). While the DCM extract showed low level of antibacterial activity against Bacillus subtilis and ampicillin-resistant Escherichia coli, the MeOH extract was active against B. cereus, B. subtilis, E. coli and ampicillin-resistant E. coli. However, the minimum inhibitory concentrations (MIC) of the MeOH extract against these bacterial strains were >10 mg/ml. None of the extracts showed any significant toxicity towards brine shrimps ($LD_{50}$ = > 1.00 mg/ml).

• 한국자기학회지
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• 제16권4호
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• pp.216-220
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• 2006

10개의 다중 홀(Hall)소자를 이용하여 요골동맥에 놓여진 영구자석의 위치 변화에 따른 전압의 변화를 전기적 신호로 얻었다.전기적 신호는 맥의 파형 신호를 의미하며 이들을 다시 회로의 하드웨어를 통해 신호들을 미분하여 자장의 변화 신호만 얻었다.소프트웨어를 통해 위치 변화에 따른 자장 변화의 크기에 따라 그 형태를 3차원 영상을 얻어 착용형 맥진기로서의 개발 가능성을 보여주었다.