• 생약학회지
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• 제47권3호
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• pp.264-272
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• 2016

This study was carried out to evaluate the preventive effect of the Corni Fructus (SSU) 50 % EtOH extract (SSU-E50) against bisphenol A (BPA) toxicity in Leydig cells and improving testosterone deficiency syndrome in orchidectomized Sprague-Dawly (SD) rats. Antioxidant properties were measured by radical scavenging activity of SSU-E50 in ABTS assay and DPPH assay. Also, real-time polymerase chain reaction(real-time PCR) was performed to quantify the mRNA expression levels of antioxidant enzyme. SD rats were divided into eight group: normal, sham operation (Sham), orchidectomized (ORX), ORX treated with testosterone 1 mg/kg (Tes. 1), ORX treated with SSU water extract 100 mg/kg (SSU-A 100) and 300 mg/kg (SSU-A 300), ORX treated with SSU 50 % EtOH extract 100 mg/kg (SSU-E 100) and 300 mg/kg (SSU-E 300). On a comparative basis, the SSU showed better activity quenching ABTS with an IC50 value of 0.29 mg/ml and DPPH with an IC50 value of 0.33 mg/ml. Cell viability was evaluated by MTS assay as described not cytotoxic at the highest concentration of $500{\mu}g/ml$. Cytotoxicity of BPA showed in $200{\mu}M$, but definitely survived by treatment with SSU in Leydig cells. In addition, SSU increased the mRNA expression levels of antioxidant enzyme in BPA induced Leydig cells. Superoxide dismutase (SOD) level was slightly increased and malondialdehyde (MDA) level was decreased with SSU-A 100 in in-vivo. These results suggest that Corni Fructus extracts have the greatest property as a natural anti-oxidative and improves testosterone deficiency syndrome source.

• 대한한의학방제학회지
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• 제19권1호
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• pp.207-217
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• 2011

Objectives : In a previous study, we have shown that Gagam-Gongjin-Dan(GGD) has an inhibitory effect on the ovalbumin-induced immune responses and a hepatoprotective effect on actaminophen-induced liver injury in Balb/c Mice. However, the possible anti-inflammatory effect of GGD extract for inflammatory mediators was not reported. Therefore, the purpose of this study was to investigate an inhibitory effects of GGD extract against lipopolysaccharides(LPS) induced inflammatory mediators in mouse peritoneal macrophages. Methods : GGD extract was prepared by extracting with methanol for 7 days. The extract was freeze-dried following filtration through vacuum distillation system. Accumulated nitrite, an oxidative product of nitric oxide(NO), was measured in the culture medium by the Griess reaction. The levels of prostaglandin $E_2(PGE_2)$, interleukin-$1{\beta}$(IL-$1{\beta}$), tumor necrosis factor-${\alpha}$(TNF-${\alpha}$) were measured by enzyme-linked immunosorbent assay. The expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2(COX-2) were measured by Western blot analysis. Results : GGD extract (50-$400\;{\mu}g$/ml) per se had no cytotoxic effect in LPS-stimulated peritoneal macrophages. GGD extract dose-dependently reduced NO, $PGE_2$, IL-$1{\beta}$ and TNF-${\alpha}$ production and COX-2 activity caused by stimulation of LPS. The levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with GGD extract in a dose dependent manner. Conclusions : These results suggest that GGD extract has an anti-inflammatory effect against LPS-induced inflammatory mediators in peritoneal macrophages, these properties may contribute to inflammation disease care.

• Clinical and Experimental Reproductive Medicine
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• 제29권4호
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• pp.323-335
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• 2002

Object: The present study was accomplished to obtain a gene expression profile of the luminal epithelium during embryo apposition in comparison of implantation (1M) and interimplantation (INTER) sites. Material and Method: The mouse uterine luminal epithelium from IM and INTER sites were sampled on day 4.5 (Day of vaginal plug = day 0.5) by Laser Captured Microdissection (LCM). RNA was extracted from LCM captured epithelium, amplified, labeled and hybridized to microarrays. Results from microarray hybridization were analyzed by Significance Analysis of Microarrays (SAM) method. Differential expression of some genes was confirmed by LCM followed by RT-PCR. Results: Comparison of IM and INTER sites by SAM identified 73 genes most highly ranked at IM, while 13 genes at the INTER sites, within the estimated false discovery rate (FDR) of 0.163. Among 73 genes at IM, 20 were EST/unknown function, and the remain 53 were categorized to the structural, cell cycle, gene/protein expression, immune reaction, invasion, metabolism, oxidative stress, and signal transduction. Of the 24 structural genes, 14 were related especially to extracellular matrix and tissue remodeling. Meanwhile, among 13 genes up-regulated at INTER, 8 genes were EST/unknown function, and the rest 5 were related to metabolism, signal transduction, and gene/protein expression. Among these 58 (53+5) genes with known functions, 13 genes (22.4%) were related with $Ca^{2+}$ for their function. Conclusions: Results of the present study suggest that 1) active tissue remodeling is occurring at the IM sites during embryo apposition, 2) the INTER sites are relatively quiescent than IM sites, and 3) the $Ca^{2+}$ may be a crucial for apposition. Search for human homologue of those genes expressed in the mouse luminal epithelium during apposition will help to understand the implantation process and/or implantation failure in humans.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2013년도 제44회 동계 정기학술대회 초록집
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• pp.108-109
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• 2013

Mitochondria play key roles in the production of cell's energy. Their dominant function is the synthesis of adenosine 5'-triphosphate (ATP) from adenosine diphosphate (ADP) and phosphate (Pi) through the oxidative phosphorylation. Evaluation of drug-induced mitochondrial toxicity has become increasingly important since mitochondrial dysfunction has recently been implicated in numerous diseases including cancer and diabetes mellitus. Mitochondrial functions have been monitored via oxygen consumption, mitochondrial membrane potential, and more importantly via ATP synthesis since ATP synthesis is the most essential function of mitochondria. Various analytical methods have been employed to investigate ATP synthesis in mitochondria, including high performance liquid chromatography (HPLC), bioluminescence technique, and pH measurement. However, most of these methods are based on destructive analysis or indirect monitoring through the enzymatic reaction. Infrared absorption spectroscopy (IRAS) is one of the useful techniques for real-time, label-free, and direct monitoring of biological reactions [1,2]. However, the strong water absorption requires very short path length in the order of several micrometers. Transmission measurements with thin path length are not suitable for mitochondrial assays because solution handlings necessary for evaluating mitochondrial toxicity, such as rapid mixing of drugs and oxygen supply, are difficult in such a narrow space. On the other hand, IRAS in the multiple internal reflection (MIR) geometry provides an ideal optical configuration to combine solution handling and aqueous-phase measurement. We have recently reportedon a real-time monitoring of drug-induced necrotic and apoptotic cell death using MIR-IRAS [3,4]. Clear discrimination between viable and damaged cells has been demonstrated, showing a promise as a label-free and real-time detection for cell-based assays. In the present study, we have applied our MIR-IRAS system to mitochondria-based assays by monitoring ATP synthesis in isolated mitochondria from rat livers. Mitochondrial ATP synthesis and hydrolysis were in situ monitored with MIR-IRAS, while dissolved oxygen level and solution pH were simultaneously monitored with O2 and pH electrodes, respectively. It is demonstrated that ATP synthesis and hydrolysis can be monitored by the IR spectral changes in phosphate groups in adenine nucleotides and MIR-IRAS is useful for evaluating time-dependent drug effects of mitochondrial toxicants.

• 한국식품영양과학회지
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• 제21권6호
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• pp.693-700
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• 1992

도토리의 천연 항산화제로서의 이용성을 검토하기 위하여 도토리 분말로부터 수종의 유기용매를 가하여 항산화성 획분을 추출 분리한 후 대두유, 팜유, 돈지 및 우지에 첨가하고 $60^{\circ}C$에서 저장하면서 POV의 변화를 측정하였으며, 항산화 성분을 TLC 및 HPLC로 분리 동정하였다. 도토리의 일반성분은 수분 11.8~12.0%, 단백질 7.1~7.4%, 전분 65.5~69.4%, 조지방 2.1~2.6%, 조섬유 2.1~3.6% 및 조회분 2.4~2.6%이었으며 total tannin 함량은 4.6~6.8%이었다. 도토리류 분말로부터 acetone : water 및 ethyl acetate를 차례로 사용하여 항산화성 획분을 추출하여 얻어진 추출물의 수율은 2.8~3.1%이었다. 최종단계의 도토리 추출물 중에는 gallic acid, digallic acid 및 gallotannin이 함유되어 있었다. 도토리 추출물의 항산화성분은 주로 gallic acid이었다. 도토리 추출물은 유탁액상태의 기질에서는 강한 항산화력을 나타내었고, 유지에 직접 첨가하였을 때는 항산화력이 인정되지 않았다. 우지 및 대두유탁액에 대산 도토리 추출물의 항산화 효과는 강하였으나 돈지 및 팜유유탁액에 대해서는 약하였다. 도토리 추출물은 0.02%의 농도에서 실용적인 항산화 효과가 인정되었다.

• Animal Bioscience
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• 제34권10호
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• pp.1590-1599
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• 2021

Objective: This study investigates the expression patterns of toll-like receptors (TLRs) and intracellular mediators in horse muscle cells after exercise, and the relationship between TLRS expression in stressed horse muscle cells and immune cell migration toward them. Methods: The expression patterns of the TLRs (TLR2, TLR4, and TLR8) and downstream signaling pathway-related genes (myeloid differentiation primary response 88 [MYD88]; activating transcription factor 3 [ATF3]) are examined in horse tissues, and horse peripheral blood mononuclear cells (PBMCs), polymorphonuclear cells (PMNs) and muscles in response to exercise, using the quantitative reverse transcription-polymerase chain reaction (qPCR). Expressions of chemokine receptor genes, i.e., C-X-C motif chemokine receptor 2 (CXCR2) and C-C motif chemokine receptor 5 (CCR5), are studied in PBMCs and PMNs. A horse muscle cell line is developed by transfecting SV-T antigen into fetal muscle cells, followed by examination of muscle-specific genes. Horse muscle cells are treated with stressors, i.e., cortisol, hydrogen peroxide (H₂O₂), and heat, to mimic stress conditions in vitro, and the expression of TLR4 and TLR8 are examined in stressed muscle cells, in addition to migration activity of PBMCs toward stressed muscle cells. Results: The qPCR revealed that TLR4 message was expressed in cerebrum, cerebellum, thymus, lung, liver, kidney, and muscle, whereas TLR8 expressed in thymus, lung, and kidney, while TLR2 expressed in thymus, lung, and kidney. Expressions of TLRs, i.e., TLR4 and TLR8, and mediators, i.e., MYD88 and ATF3, were upregulated in muscle, PBMCs and PMNs in response to exercise. Expressions of CXCR2 and CCR5 were also upregulated in PBMCs and PMNs after exercise. In the muscle cell line, TLR4 and TLR8 expressions were upregulated when cells were treated with stressors such as cortisol, H₂O₂, and heat. Migration of PBMCs toward stressed muscle cells was increased by exercise and oxidative stresses, and combinations of these. Treatment with methylsulfonylmethane (MSM), an antioxidant on stressed muscle cells, reduced migration of PBMCs toward stressed muscle cells. Conclusion: In this study, we have successfully cultured horse skeletal muscle cells, isolated horse PBMCs, and established an in vitro system for studying stress-related gene expressions and function. Expression of TLR4, TLR8, CXCR2, and CCR5 in horse muscle cells was higher in response to stressors such as cortisol, H₂O₂, and heat, or combinations of these. In addition, migration of PBMCs toward muscle cells was increased when muscle cells were under stress, but inhibition of reactive oxygen species by MSM modulated migratory activity of PBMCs to stressed muscle cells. Further study is necessary to investigate the biological function(s) of the TLR gene family in horse muscle cells.

• 대한통합의학회지
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• 제9권2호
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• pp.83-92
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• 2021

Purpose : Keratinocytes are the main cellular components involved in wound healing during re-epithelization and inflammation. Dysfunction of tight junction (TJ) adhesions is a major feature in the pathogenesis of various diseases. The purpose of this study was to identify the various effects of a Sparassis crispa water extract (SC) on HaCaT cells and to investigate whether these effects might be applicable to human skin. Methods : We investigated the effectiveness of SC on cell HaCaT viability using MTS. The antioxidant effect of SC was analyzed by comparing the effectiveness of ABTS to that of the well-known antioxidant resveratrol. Reverse-transcription quantitative polymerase chain reaction (qRT-PCR) is the most widely applied method Quantitative RT-PCR analysis has shown that SC in HaCaT cells affects mRNA expression of tight-junction genes associated with skin moisturization. In addition, Wound healing is one of the most complex processes in the human body. It involves the spatial and temporal synchronization of a variety of cell types with distinct roles in the phases of hemostasis, inflammation, growth, re-epithelialization, and remodeling. wound healing analysis demonstrated altered cell migration in SC-treated HaCaT cells. Results : MTS analysis in HaCaT cells was found to be more cytotoxic in SC at a concentration of 0.5 mg/㎖. Compared to 100 µM resveratrol, 4 mg/㎖ SC exhibited similar or superior antioxidant effects. SC treatment in HaCaT cells reduced levels of claudin 1, claudin 3, claudin 4, claudin 6, claudin 7, claudin 8, ZO-1, ZO-2, JAM-A, occludin, and Tricellulin mRNA expression by about 1.13 times. Wound healing analysis demonstrated altered cell migration in SC-treated HaCaT cells and HaCaT cell migration was also reduced to 73.2 % by SC treatment. Conclusion : SC, which acts as an antioxidant, reduces oxidative stress and prevents aging of the skin. Further research is needed to address the effects of SC on human skin given the observed alteration of mRNA expression of tight-junction genes and the decreased the cell migration of HaCaT cells.

• Korean Journal of Acupuncture
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• 제36권1호
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• pp.63-73
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• 2019

Objectives : This study was designed to investigate the effects of Alismatis Rhizoma (AR) pharmacopuncture at BL23 on acute renal failure induced by cisplatin. Methods : The Sprague-Dawley rats were divided into Normal group (no injection of cisplatin and no treatment), Control group (cisplatin injection without treatment), Acu group (needling at BL23 after cisplatin injection), AR-PA1 group (treated with $0.3571mg/kg/20{\mu}l$ of AR pharmacopuncture at BL23 after cisplatin injection), and AR-PA2 group (treated with $1.7855mg/kg/20{\mu}l$ of AR pharmacopuncture at BL23 after cisplatin injection). Each treatment was given once daily for 8 days. Changes in body weight, kidney weight, tumor necrosis factor-alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), Cu-Zn superoxide dismutase (Cu-Zn SOD), glutathione peroxidase (GPx), serum blood urea nitrogen (serum BUN), and serum creatinine were observed. Results : Body weight was significantly increased in AR-PA1 on $4^{th}$ and $6^{th}$ days and AR-PA2 on $2^{nd}$ day. $TNF-{\alpha}$ was significantly decreased in Acu, AR-PA1 and AR-PA2 groups. Cu-Zn SOD was significantly increased in AR-PA2 group. GPx was significantly increased in AR-PA1 and AR-PA2 groups. But kidney weight, IL-6, serum BUN and serum creatinine were not significantly changed in any groups compared to control group. Conclusions : In acute renal failure induced by cisplatin, AR pharmacopuncture has a mitigating effect on the inflammatory reaction related to the increase of $TNF-{\alpha}$ in the kidney tissue and a protective effect on the oxidative stress of the kidney tissue. However it is unlikely to restore the glomerular function or inhibit the renal swelling.

• 공업화학
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• 제32권4호
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• pp.461-466
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• 2021

본 논문에서는 이동성이 좋고 경제적이며, 간편하게 일회용 진단칩으로 제작 가능한 스크린 프린팅 한 탄소칩 전극[screen printed carbon electrode (SPCE)] 기반의 전압전류법 나노물질 융합형 바이오센서를 제작하여 폐암 조기진단에 활용 가능한 단백질 표지 인자 중에 하나인 heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) 단백질의 농도를 정량 분석하고자 하였다. 먼저 SPCE 표면에 금 나노입자를 전기적으로 증착한 후 크로스링커를 이용하여 hnRNP A1에 특이적으로 결합할 수 있는 바이오리셉터인 DNA 압타머를 고정하였다. Ethanolamine을 블로킹 시약으로 사용하여 압타머와 함께 센서 표면에 고정하여 그 표면을 처리함으로써 비특이적인 생물질의 흡착에 의한 방해 신호를 최소화하고자 하였다. DNA칩과 hnRNP A1 용액을 접촉하여 DNA와 hnRNP A1을 결합시킨 후 alkaline phosphatase (ALP) 효소로 접합한 hnRNP A1 항체(anti-hnRNP A1)을 센서칩 표면으로 주입하여 샌드위치 복합체를 형성하고, 이를 기질인 4-aminophenyl phosphate (APP)와 효소-기질 특이적 산화 반응에 의한 전류 변화를 순환 전압전류법과 시차 펄스전압전류법으로 측정하여 단백질의 농도를 정량적으로 분석하였다. 상기 산화 반응에 의한 피크 전류 변화는 순환전압전류법과 시차 펄스 전압전류법을 사용할 때 -0.05와 -0.17 V (vs. Ag/AgCl) 전위 값에서 각각 일어났다. 개발한 나노바이오센서를 실제 정상인 혈청 시료 분석에 적용 가능함을 보여줌으로써 혈청 한 방울로 폐암의 조기진단 가능성을 제시하고자 하였다.

• 생명과학회지
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• 제32권1호
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• pp.63-69
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• 2022

Thioredoxin reductase (TrxR) 시스템은 세포 내 산화 환원 반응의 항상성 유지와 신호 전달 경로를 조절하는데 중추적인 역할을 함으로써 세포의 생존과 기능 유지에 필수적이다. TrxR 시스템은 thioredoxin (Trx), TrxR 및 nicotinamide adenine dinucleotide phosphate의 구성요소를 포함하며, TrxR 효소의 촉매 반응에 의해 환원된 Trx는 하위 표적 단백질을 환원시켜 결과적으로 산화적 스트레스에 대한 방어와 세포 분화, 성장 및 사멸을 조절한다. 암세포는 무한한 세포 증식과 높은 대사율로 인해 과도하게 생성된 활성산소종을 소거하기 위해 세포 내 항산화능을 향상시켜 세포의 생존을 유지하는 반면, 항산화 시스템에 대한 의존도 및 민감도가 높아 이를 표적으로 한 항암 활성 연구에서 잠재적인 가능성이 있음을 제시한다. 여러 연구 결과에서 TrxR이 다양한 유형의 암에서 높은 수준으로 발현되고 있음이 밝혀졌고, 또한 TrxR 시스템을 표적으로 한 항암 활성에 대한 연구가 증가하고 있다. 따라서 본 총설에서는 세포 내 TrxR 시스템의 기능과 암의 발달 및 진행에서의 역할을 다루고, TrxR 억제제의 항암 활성 및 기전을 검토함으로써 항암 활성 연구에 대한 전략으로 TrxR 시스템의 타당성과 가치를 논하였다.