• 생명과학회지
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• 제26권12호
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• pp.1431-1437
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• 2016

본 연구에서는 수수 에탄올 추출물(ethanol extract of Sorghum bicolor L., SE)을 이용하여 RC-58T/h/SA#4 primary 인체 전립선 암세포에 대한 apoptosis 유도효과 및 RAW 264.7 마우스 대식세포에 대한 면역활성 증진효능에 대해 알아보고자 하였다. SE의 처리는 RC-58T/h/SA#4 세포의 증식을 억제하였으며 세포수축을 통한 형태학적 변화를 유발하였다. 또한 apoptosis 관련 지표현상인 apoptotic body 및 핵의 형태변화를 관찰할 수 있었다. RC-58T/h/SA#4 세포에 SE를 처리하였을 때 caspase-8, -9, -3 단백질 활성을 증가시켰으며 pro-apoptotic 단백질인 Bax, p53, 분절된 PARP 및 세포질의 cytochrome c 단백질은 증가시킨 반면, anti-apoptotic 단백질인 Bcl-2 단백질은 감소시켰다. 이는 z-VAD-fmk로 caspase 활성을 억제시켰을 때 SE에 의한 세포증식 억제효과가 감소되었으며 이를 통해 SE에 의한 세포증식 억제효과는 caspase 의존적임을 확인할 수 있었다. 면역반응에서 nitric oxide(NO)의 증가는 면역활성 지표로 여겨지며, 본 연구에서 SE를 처리하였을 때 RAW 264.7 세포에서 NO 생성능이 증가하는 것으로 나타났다. 본 연구의 결과들은 SE가 primary 전립선 암세포에서 caspase 의존형 apoptosis를 유도하며 대식세포의 면역활성을 증가시킨다는 것을 나타낸다. 따라서 수수를 전립선 암 예방 및 면역활성을 증진효능이 있는 기능성 식품 소재로 활용할 수 있을 것으로 생각된다.

• 한국자원식물학회지
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• 제25권2호
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• pp.232-239
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• 2012

서양민들레의 부위별 성분 및 생리활성 차이를 검토하고자 폴리페놀 함량, 플라보노이드 함량, 항산화성 및 세포독성을 분석하였다. Folin-Denis방법에 따른 총 페놀 함량은 꽃 추출물에서 72.0 mg $kg^{-1}$으로 가장 높게 나타났으며, 그 다음이 잎, 뿌리, 꽃대 추출물 순으로 나타났다(p<0.05). 한편, 총 플라보노이드 함량은 총 페놀 함량과 같은 경향을 보였으나 더 낮은 범위의 함량이 검출되었다. DPPH 라디컬 소거능은 추출물 농도가 증가할수록 높은 활성을 보였으며 그 중 꽃 추출물 ($IC_{50}$ 값 = 624.3 mg $kg^{-1}$)에서 가장 높은 활성을 보였고 그 다음으로 잎, 뿌리, 꽃대 추출물 순으로 나타나($p$ <0.05), 이는 페놀 및 플라보노이드 함량이 DPPH 라디컬 소거능과 관련이 있음을 보여 주었다. MTT법에 의한 세포독성 시험에서 폐암세포주(Calu-6)와 위암세포주(SNU-601)의 세포 생존율은 꽃 추출물에서 가장 낮았고($IC_{50}$값 = 85.7과 311.4 mg $kg^{-1}$), 그 다음으로 잎, 꽃대, 뿌리 추출물 순으로 뿌리 추출물이 가장 높은 생존율을 보였다. 이는 세포독성이 꽃 추출출물에서 가장 높고, 뿌리에서 가장 낮은 것을 보여 준 것이다. 결론적으로 총 페놀 함량과 총 플라보노이드 함량이 항산화성($r^2$=0.7280 to 0.9971)과 세포독성($r^2$=0.5795 to 0.9515)에 높은 상관관계가 있고, 그 함량과 활성은 식물체 부위별로 다르게 나타남을 확인하였다.

• 대한화장품학회지
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• 제49권1호
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• pp.87-96
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• 2023

본 연구는 인간 각질형성 세포주를 이용하여 피부 표면의 산성화를 조절하는 물질을 발굴하고, 이를 통해 각질층의 보습 능력 및 피부 장벽 기능에 미치는 영향을 조사하기 위해 수행되었다. 꿀풀은 아프리카 북서부 및 북미에 널리 분포하는 허브 중 하나로 세포사멸, 항산화 및 항염에 대한 효능이 연구되어 왔다. 하지만 NHE1의 발현 조절 및 피부 장벽기능 회복에 대한 연구는 진행되지 않았다. 꿀풀의 피부 pH 조절 효과를 확인하기 위하여 꿀풀의 활성 성분 분석 결과 로즈마린산과 카페인산이 검출되었다. 꿀풀과 이것으로부터 검출된 성분인 카페인산은 인간 각질형성 세포주인 HaCaT 세포에 고농도(100 ㎍/mL 또는 100 µM)를 처리한 결과 독성을 보이지 않았다. 노화된 피부에서는 각질층의 산성 pH를 유지하기 위한 나트륨-수소 이온 교환 펌프(NHE1)의 발현감소가 나타나는 것으로 알려져 있으며, 이러한 이유로 노화 피부에서 나타나는 피부 장벽 기능 회복의 저하에 중요한 원인으로 작용할 것으로 추측된다. 꿀풀 추출물 및 카페인산은 각질형성 세포에서 NHE1의 발현을 증가시켰으며, 자연 보습 인자 전구체인 필라그린과 세라마이드 합성 효소인 serine plmitoyl transferase (SPT)의 발현을 증가시켰다. 또한, 직접적인 pH 조절 효과를 확인하기 위해 각질형성 세포 내/외 pH 수치를 측정한 결과 꿀풀과 카페인산은 세포 외부 pH를 감소시켰다. 위 결과들을 종합해 볼 때, 꿀풀과 카페인산은 NHE1 조절을 통해 피부 pH를 조절할 수 있고, 자연 보습 인자(NMF)와 세라마이드 합성을 증가시켜 피부 장벽 기능 회복을 도울 수 있을 것으로 추측된다. 이러한 결과는 꿀풀과 카페인산이 피부 건강에 긍정적인 영향을 미칠 수 있다는 가능성을 보여주며, 이를 활용한 새로운 피부 보호 제품 개발에 대한 토대가 될 수 있다.

• Tuberculosis and Respiratory Diseases
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• 제47권4호
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• pp.471-477
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• 1999

연구배경 : 발암물질의 체내 대사효소계인 GSTM1, T1과 NAT1의 유전적다형성의 발현 양상이 폐암발생과 연관성이 있는가를 밝히기 위해 환자-대조군 연구를 수행하였다. 방 법 : 서울대병원에서 병리화적 폐암환자군으로, 비암성요료계질환으로 입원한 환자를 대조군으로 설정하여, 각 환자에 대한 성, 연령, 흡연력 등을 병력조사나 병록지검토로 얻었으며, 말초혈액을 채취하여 DNA를 분리한 뒤, 다중중합효소연쇄반응을 이용하여 GSTM1, T1의 유전자형을, nested PCR법을 이용하여 NAT1의 유전자형을 결정하였다. 결 과 : 모집된 환자, 대조군은 각각 118 명, 150명이었고, 환자군에서 흡연력은 통계적으로 의미있게 높았다(p<0.05). GSTM1의 소실형은 비소실형에 비해 편평상피암의 위험을 높였으나(OR=2.25 ; 95% CI=1.12-4.51), GSTT1의 경우는 폐암의 위험인자로 작용하지 않았다. NAT1의 fast acetylator형은 폐암환자전체를 분석했을 때 통계적으로 의미있는 위험도 상승을 보였다(OR=2.13; 95% CI=1.04-4.40). 결 론 : GSTM1의 소실형과 NAT1의 fast형은 폐암발생과 관련성이 있다.

• 동의생리병리학회지
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• 제27권4호
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• pp.400-408
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• 2013

This study aimed to investigate the relaxation effects and its underlying mechanisms of Rubus coreanus(RC) extract in contracted rabbit corpus cavernous tissues by phenylephrine(PE) $1{\mu}M$. In order to define the relaxation effects of RC, rabbit corpus cavernous tissues were prepared in $2{\times}2{\times}6mm$ sized strip. The dose-dependent relaxation responses of RC at 0.01-3.0 $mg/m{\ell}$ in contracted strips induced by PE were measured and also observed after endothelial denudation. To analyze the underlying mechanisms of RC-induced relaxation, indomethacin(IM), tetraethylammonium chloride(TEA), $N{\omega}$-nitro-L-arginine (L-NNA), methylene blue(MB) were treated before RC extract infused into precontracted strips induced by PE. To study the effect of RC extract on influx of extracellular $Ca^{2+}$ in corpus cavernous strips, calcium chloride(Ca) 1 mM infused into precontracted strips induced by PE after pretreatment of RC extract in $Ca^{2+}$-free krebs-ringer solution. To investigate cytotoxic activity and nitric oxide(NO) concentration of RC extract on human umbilical vein endothelial cell(HUVEC), cell viability on HUVEC was measured by MTT assay, and NO concentration was measured by Griess reagent system. The cavernous strips were significantly relaxed by RC extract at 1.0 $mg/m{\ell}$, 3.0 $mg/m{\ell}$ and the relaxation responses to RC were inhibited significantly by endothelial disruption. The pretreatment of IM, TEA didn't affect RC extract-induced endothelium-dependent relaxation, but the pretreatment of L-NNA, MB reduced RC extract-induced endothelium-dependent relaxation. When $Ca^{2+}$ was supplied the cavernous strips which were precontracted by PE in a $Ca^{2+}$-free krebs-ringer solution, contraction of strips was increased, but pretreatment of RC inhibited contractile response to $Ca^{2+}$. When RC extract was applicated on HUVEC, NO concentration was increased. Our findings show that RC extract exerts a relaxing effect on corpus cavernosum in part by suppressing influx of extracellular $Ca^{2+}$ through activating the NO-cGMP system.

• Clinical and Experimental Pediatrics
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• 제55권11호
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• pp.438-444
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• 2012

Mucolipidosis II (ML II) or inclusion cell disease (I-cell disease) is a rarely occurring autosomal recessive lysosomal enzyme-targeting disease. This disease is usually found to occur in individuals aged between 6 and 12 months, with a clinical phenotype resembling that of Hurler syndrome and radiological findings resembling those of dysostosis multiplex. However, we encountered a rare case of an infant with ML II who presented with prenatal skeletal dysplasia and typical clinical features of severe secondary hyperparathyroidism at birth. A female infant was born at $37^{+1}$ weeks of gestation with a birth weight of 1,690 g (<3rd percentile). Prenatal ultrasonographic findings revealed intrauterine growth retardation and skeletal dysplasia. At birth, the patient had characteristic features of ML II, and skeletal radiographs revealed dysostosis multiplex, similar to rickets. In addition, the patient had high levels of alkaline phosphatase and parathyroid hormone, consistent with severe secondary neonatal hyperparathyroidism. The activities of ${\beta}$-D-hexosaminidase and ${\alpha}$-N-acetylglucosaminidase were moderately decreased in the leukocytes but were 5- to 10-fold higher in the plasma. Examination of a placental biopsy specimen showed foamy vacuolar changes in trophoblasts and syncytiotrophoblasts. The diagnosis of ML II was confirmed via GNPTAB genetic testing, which revealed compound heterozygosity of c.3091C>T (p.Arg1031X) and c.3456_3459dupCAAC (p.Ile1154GlnfsX3), the latter being a novel mutation. The infant was treated with vitamin D supplements but expired because of asphyxia at the age of 2 months.

• Asian Pacific Journal of Cancer Prevention
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• 제14권1호
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• pp.201-208
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• 2013

Objective: To investigate changes in the invasive capacity of gastric cancer cells in vitro after expression inhibition of T lymphoma invasion and metastasis inducing factor 1 (Tiam 1) and underlying mechanisms. Methods: Using adhesion selection, two subpopulations with high ($M_H$) or low ($M_L$) invasive capacity were separated from the human gastric cancer cell line MKN-45 ($M_0$). Tiam 1 antisense oligodeoxynucleotide (ASODN) was transfected into $M_H$ cells with liposomes, and expression of Tiam 1 mRNA and protein was determined by RT-PCR and quantitative cellular-ELISA. Changes in the cytoskeleton, invasive capacity in vitro and expression of ras-related $C_3$ botulinum toxin substrate 1 (Rac 1), integrin ${\beta}1$ and matrix metalloproteinase 2 (MMP 2) between Tiam 1 ASODN transfected $M_H$ cells and non-transfected cells were observed by HE staining, cytoskeletal protein staining, scanning electron microscopy, Boyden chamber tests and cyto-immunohistochemistry. Results: A positive correlation existed between the expression level of Tiam l mRNA or protein and the invasion capacity of gastric cancer cells. After ASODN treatment ($0.43{\mu}M$ for 48 h), Tiam 1 mRNA transcription and protein expression in $M_H$ cells were decreased by 80% and 24% respectively (P < 0.05), compared with untreated controls, while invasive capacity in vitro was suppressed by 60% (P < 0.05). Morphologic and ultrastructural observation also showed that ASODN-treated $M_H$ cells exhibited smooth surfaces with obviously reduced filopodia and microspikes, which resembled $M_0$ and $M_L$ cells. Additionally, cytoskeletal distribution dramatically altered from disorder to regularity with reduced long filament-like structure, projections, pseudopodia on cell surface, and with decreased acitn-bodies in cytoplasm. After Tiam 1 ASODN treatment, the expression of Rac 1 and Integrin ${\beta}1$ in $M_H$ cells was not affected (P > 0.05), but that of MMP 2 in $M_H$ cells was significantly inhibited compared with untreated cells (P < 0.05). Conclusion: Over-expression of Tiam-1 contributes to the invasive phenotype of gastric cancer cells. Inhibition of Tiam 1 expression could impair the invasive capacity of gastric cancer cells through modulating reconstruction of the cytoskeleton and regulating expression of MMP 2.

• KSBB Journal
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• 제20권4호
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• pp.278-284
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• 2005

식물세포의 느린 생장과 낮은 생산량의 이유로 지금까지 는 주로 미생물이나 동물세포에서 유전자 재조합 단백질을 생산하여 왔다. 그러나 저렴한 배지 가격, 동물 유래 바이러스 감염 위험성으로부터의 안정성, glycosylation 등의 post-translational modification이 가능하다는 장점들로 인하여 최근 들어 식물세포배양은 생물학적 활성을 가진 고부가가치의 단백질을 생산하는데 많이 이용되고 있다. 본 연구에서는 생장배지에 첨가했던 sucrose의 소비와 induction 배지로의 교환에서 오는 배지내의 삼투압을 조절하여 hCTLA4-Ig의 생산성을 높이고자 하였다. 다양한 삼투압 조절제 첨가 실험을 통해 sorbitol을 선별하고, 40 mM의 sorbitol 첨가에서 상대적으로 높은 생존도와 induction 후 7일째 대조구보다 1.7배 높은 생산성을 확인하였다. 또한, 저농도의 glucose 첨가를 통한 생산성 증대에 있어서는 8 mM glucose에서 induction 이후에도 높은 세포농도를 유지하면서 최대 37.3 mg/L까지 hCTLA4-Ig 생산량을 증가시켰다. 5-L bioreactor에서 회분식 배양과 induction시의 hCTLA4-Ig 생산량을 비교한 결과 induction시 배양 18일째 최고 45.3 mg/L까지 높일 수 있었으며, 회분식 배양에 비해 2.1배 증가됨을 확인하였다.

• Clinical and Experimental Reproductive Medicine
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• 제29권1호
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• pp.21-28
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• 2002

Objective: To evaluate whether co-culture of oocytes on vero cell monolayers from Day 0 (Day 0 group) after egg retrieval results in an increase in developmental capacity such as fertilization rate, embryo quality, blastulation and clinical pregnancy rate compared with co-culture of oocytes from Day 1 (Day 1 group). Methods: Sperms were treated with Hams F-10 supplemented with 10% human follicular fluid (hFF). Vero cells for co-culture were prepared in TCM-199 with 10% FBS. Oocytes were co-cultured from Day 0 and fertilized oocytes were co-cultured from Day 1 on vero cell monolayers in DMEM with 10% and 20% hFF, respectively after egg retrieval. On day 1, 2 and 5, fertilization rate and grade of embryos and blastocysts were evaluated. Results (fertilization rate, cleavage rate, grade of embryos and blastocysts and pregnancy rate) were considered statistically significant when p value was less than 0.05 using t-test and $x^2$. Results: In sibling oocytes of same cycles, no differences were found in fertilization rate (94.6 vs. 91.4%), cleavage rates (94.6 vs. 91.4%), embryo grade (on day 2 and 3) and blastulation (65.6 vs. 57.0%) and their grade. In different oocytes of different cycles (patients), no differences were found in fertilization (79.8 vs. 78.3%), cleavage rates (77.7 vs. 76.4%) and blastulation (56.0 vs. 45.3%), but pregnancy rate was higher in the Day 0 group than in the Day 1 group (60.0 vs. 42.9%). Conclusions: This study revealed that the embryonic development capacities were not affected by the different co-culture time in the sibling oocytes of same cycles. Although no statistical significance, because of small size of study, there was a trend for higher pregnancy rates in Day 0 group compared to Day 1 group in different oocytes of different cycles.

• Asian-Australasian Journal of Animal Sciences
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• 제31권4호
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• pp.595-606
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• 2018

Objective: The Fusarium mycotoxins of deoxynivalenol (DON) and zerolenone (ZEN) cause health hazards for both humans and farm animals. Therefore, the main intention of this study was to reveal DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the liver of piglets. Methods: In the present study, 15 six-week-old piglets were randomly assigned to the following three different dietary treatments for 4 weeks: control diet, diet containing 8 mg DON/kg feed, and diet containing 0.8 mg ZEN/kg feed. After 4 weeks, liver samples were collected and sequenced using RNA-Seq to investigate the effects of the mycotoxins on genes and gene networks associated with the immune systems of the piglets. Results: Our analysis identified a total of 249 differentially expressed genes (DEGs), which included 99 upregulated and 150 downregulated genes in both the DON and ZEN dietary treatment groups. After biological pathway analysis, the DEGs were determined to be significantly enriched in gene ontology terms associated with many biological pathways, including immune response and cellular and metabolic processes. Consistent with inflammatory stimulation due to the mycotoxin-contaminated diet, the following Kyoto encyclopedia of genes and genomes pathways, which were related to disease and immune responses, were found to be enriched in the DEGs: allograft rejection pathway, cell adhesion molecules, graft-versus-host disease, autoimmune thyroid disease (AITD), type I diabetes mellitus, human T-cell leukemia lymphoma virus infection, and viral carcinogenesis. Genome-wide expression analysis revealed that DON and ZEN treatments downregulated the expression of the majority of the DEGs that were associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9), proliferation (insulin-like growth factor 1, major facilitator superfamily domain containing 2A, insulin-like growth factor binding protein 2, lipase G, and salt inducible kinase 1), and other immune response networks (paired immunoglobulin-like type 2 receptor beta, Src-like-adaptor-1 [SLA1], SLA3, SLA5, SLA7, claudin 4, nicotinamide N-methyltransferase, thyrotropin-releasing hormone degrading enzyme, ubiquitin D, histone $H_2B$ type 1, and serum amyloid A). Conclusion: In summary, our results demonstrated that high concentrations DON and ZEN disrupt immune-related processes in the liver.