• Archives of Plastic Surgery
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• 제39권6호
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• pp.593-599
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• 2012

Background This study aimed to investigate the possibility of isolating mesenchymal stem cells (MSCs) from human thigh adipose tissue and the ability of human thigh adipose stem cells (HTASCs) to differentiate into hepatocytes. Methods The adipose-derived stem cells (ADSCs) were isolated from thigh adipose tissue. Growth factors, cytokines, and hormones were added to the collagen coated dishes to induce the undifferentiated HTASCs to differentiate into hepatocyte-like cells. To confirm the experimental results, the expression of hepatocyte-specific markers on undifferentiated and differentiated HTASCs was analyzed using reverse transcription polymerase chain reaction and immunocytochemical staining. Differentiation efficiency was evaluated using functional tests such as periodic acid schiff (PAS) staining and detection of the albumin secretion level using enzyme-linked immunosorbent assay (ELISA). Results The majority of the undifferentiated HTASCs were changed into a more polygonal shape showing tight interactions between the cells. The differentiated HTASCs up-regulated mRNA of hepatocyte markers. Immunocytochemical analysis showed that they were intensely stained with anti-albumin antibody compared with undifferentiated HTASCs. PAS staining showed that HTASCs submitted to the hepatocyte differentiation protocol were able to more specifically store glycogen than undifferentiated HTASCs, displaying a purple color in the cytoplasm of the differentiated HTASCs. ELISA analyses showed that differentiated HTASCs could secrete albumin, which is one of the hepatocyte markers. Conclusions MSCs were islolated from human thigh adipose tissue differentiate to heapatocytes. The source of ADSCs is not only abundant abdominal adipose tissue, but also thigh adipose tissue for cell therapy in liver regeneration and tissue regeneration.

• 한국식품영양과학회지
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• 제37권6호
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• pp.721-728
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• 2008

본 연구에서는 소장세포(Caco-2)와 대식세포(J774)를 이용하여 FPN과 DMT1의 유전자 발현에 hepcidin 펩타이드 호르몬이 미치는 영향을 알아보기 위하여 수행되었으며 그 결과를 요약하면 다음과 같다. Caco-2 세포에서 FPN과 DMT1의 mRNA 및 단백질 수준은 분화 진행에 따라 비례하여 증가하였으며, 특히 DMT1 단백질은 분화 초기에는 거의 발현되지 않다가 분화 7일째에 비로소 발현되기 시작한 후 급격히 증가하여 분화 17일째에는 7일째에 비해 단백질 수준이 10배 이상 크게 증가되었다. 분화된 Caco-2 세포에서 소변 hepcidin과 합성 hepcidin을 100 nM 농도로 24시간 동안 처리하였을 때, FPN 단백질 수준이 대조군에 비해 각각 60%와 70% 수준으로 유의하게 감소하였다. DMT1 단백질의 경우, 소변 hepcidin 100 nM 농도에서만 대조군의 55% 수준으로 유의하게 감소되었다. J774 세포에 소변 hepcidin 혹은 합성 hepcidin을 24시간 처리한 결과, 10 nM과 100 nM 농도에서 모두 대조군에 비해 FPN 단백질 수준이 유의적으로 감소하는 것으로 나타났으며, DMT1 단백질 수준도 소변 hepcidin 10 nM과 100 nM 처리에 의해 각각 대조군의 40%와 37% 수준으로 유의하게 감소하였다. 분화된 Caco-2 세포와 J774 세포에서 10 nM 혹은 100 nM 농도의 hepcidin 처리 시 DMT1 mRNA와 FPN mRNA 수준에는 영향을 미치지 않는 것으로 나타났으며, 이로 볼 때 hepcidin은 전사과정의 조절보다는 DMT1과 FPN 단백질로의 번역과정을 억제하거나 분해 속도를 촉진함으로써 이들 단백질의 수준을 낮추는 것으로 보인다. 이상의 결과는, hepcidin 펩타이드 호르몬이 DMT1 단백질과 FPN 단백질의 수준을 억제함으로써 체내 철분 대사 조절에 중요하게 관여함을 나타낸다. 특히 소장세포와 대식세포에 동시에 작용함으로써, 소장에서의 철분 흡수와 대식세포에서의 철분 방출을 효율적으로 억제하는 조절 인자로 작용할 수 있음을 제시한다. 앞으로 hepcidin의 생성 및 분비를 조절하는 요인에 대한 연구와 hepcidin이 실제 세포 내외로의 철분의 수송이 미치는 영향에 대한 기능적 연구가 계속적으로 이루어져야 할 것으로 사료된다.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제8권2호
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• pp.177-193
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• 2005

목 적: 담즙정체를 보이는 환자의 간조직에서 cDNA microarray를 이용한 유전자 발현 분석을 통 하여 담도 폐쇄증의 발생과정에 관여하는 유전자를 규명하고자 연구를 시행하였다. 방 법: 환자의 간조직은 담즙정체를 보이는 11명의 환자로부터 진단적 간생검시 얻은 간조직의 일부를 이용하였다. 이들의 원인 질환은 담도 폐쇄증이 7명, 신생아 간염 증후군이 4명이었다. 정상대조군의 간조직은 생체부분 간이식시 성인 공여자로부터 얻은 간조직의 일부를 이용하였다. 간조직에서 분리된 mRNA를 4.7K 인간유전자 cDNA microarray를 이용하여 유전자 발현의 양상을 분석하였다. 결 과: 분석한 4,700종의 유전자 중 담즙정체 환자 11명 모두의 간조직에서 발현이 증가된 유전자는 17종이 발견되었고, 발현이 감소된 유전자는 20종이 발견되었다. 담도 폐쇄증 환자와 신생아 간염 증후군 환자 사이에 49종의 유전자에서 발현의 차이가 관찰되었고, 특히 담도 폐쇄증에서는 발현이 증가하였으나 신생아 간염 증후군에서는 발현이 감소된 유전자는 24종이었으며 이들 유전자 발현의 차이에 의해서 두 군간의 감별이 가능하였다. 결 론: 신생아 간질환에서 담도 폐쇄증 환자들은 모두 일정한 양상을 보여주어 이에 대한 분석을 통하여 담도 폐쇄증의 조기 감별 진단을 할 수 있을것을 시사하였다. 또한 담도 폐쇄증에서 특이 발현을 보이는 유전자의 추가적인 기능 연구를 시행함으로써 담도 폐쇄증의 원인을 규명하는데 도움을 줄 수 있을 것으로 생각한다.

• Journal of Nutrition and Health
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• 제50권6호
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• pp.543-551
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• 2017

본 연구는 발효시킨 블루베리액이 고지방식이로 비만이 유도된 C57BL/6J mice에 미치는 항비만 효과 및 기전에 대해 평가하였다. 실험동물은 각 7마리씩 3군으로 나누어 고지방식이와 증류수를 섭취하는 HD군, 고지방식이와 블루베리액을 섭취하는 BHD군, 고지방식이와 블루베리발효액을 섭취하는 FBHD군으로 사육하였다. 경구투여량은 10 mg/kg BW/day로 설정하였고, 이는 12%의 블루베리액(2.5 g 블루베리 함유; 블루베리 함유 안토시아닌 함량은 약 3.75 mg, 총 폴리페놀은 약 10.3 mg 추정)을 나타낸다. 본 연구 결과, 블루베리액과 블루베리발효액은 체중 감소, 체지방량 감소, 간의 중성지방과 총 콜레스테롤 함량을 감소시켰다. 그리고 혈중 LDL-콜레스테롤을 낮추고, HDL-콜레스테롤을 증가시켰으며 AST 및 ALT 농도를 감소시켰다. 또한 렙틴 농도가 낮아졌으며 지방 합성 유전자 발현에서는 SREBP-1c, ACC 발현 수준이 유의적으로 낮아졌으며, 지방 산화 유전자 발현에서는 블루베리발효액의 ACOX 발현 수준이 유의적으로 증가하였다. 결론적으로 BHD군과 FBHD군이 항비만 효과를 나타냈었으며, 특히 FBHD군이 BHD군보다 간에서 총 콜레스테롤 20%, AST 9%, ALT 52%, 지방 합성 유전자 SREBP-1c 발현을 감소시켰고, 혈중 HDL-콜레스테롤 16.4%와 지방 산화 유전자 ACOX 발현을 유의적으로 증가시켰다. 따라서 본 실험의 결과, 블루베리는 고지방 섭취로 인한 콜레스테롤 및 지질변화에 감소 효과를 나타내며, 특히 발효를 통해 기능성이 더욱 증진되는 것으로 나타났다. 발효과정은 블루베리의 항비만 효과를 증진시키며 블루베리발효액은 기능성 식품으로서 이용가치가 높을 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제15권2호
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• pp.321-329
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• 2005

Ornithine decarboxylase (ODC) antizyme is a key regulatory protein in the control of cellular polyamines. We have isolated two distinct ODC antizyme cDNA clones (AZS and AZL) from a flounder (Paralichthys olivaceus) brain cDNA library. Their sequences revealed that both clones required translational frameshifting for expression. Taking + 1 frameshifting into account, AZS and AZL products were 221 and 218 amino acid residues long, respectively, and shared $83.3\%$ amino acid sequence identity. Comparison of the structure and nucleotide sequence of the antizyme genes showed that the genes were highly conserved in flounder, zebrafish, mouse, and human. A phylogenetic tree was constructed, based on the antizyme amino acid sequences from various species. The presence of the two types of antizyme mRNA species in brain, kidney, liver, and embryo was confirmed by using the reverse transcription­polymerase chain reaction (RT-PCR) and Northern blot analysis. Recombinant proteins of flounder ODC antizymes, containing His-Nus-S tag at the amino-terminus, were overexpressed as His-AZL and His-AZS fusion proteins in Escherichia coli BL21 (DE3) pLys by using the pET­44a(+) expression vector.

• 인간식물환경학회지
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• 제23권2호
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• pp.191-199
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• 2020

Noni has been used for medicinal purposes for more than 2,000 years in South Pacific Polynesia, China and India, and has been heavily ingested as an extract for its excellent antioxidant and anti-inflammatory effects. However, a recent study found that the noni extract causes digestive disorders, kidney problems, and liver diseases, which made it necessary to use it for other purposes than as an extract. In this study, we want to evaluate the potential of noni as an anti-oxidant, anti-inflammatory and anti-wrinkling agent. Methods: Noni was freeze-dried, extracted in water, and concentrated. Skin cells were treated with the noni extract for 24 hrs and then were exposed to UVB (55 mJ/cm²). After 48 hrs of incubation, pro-inflammatory cytokine, elastase, MMP-1 and type-1 procollagen levels were measured by ELISA. Results: To find out the antioxidant effect of the noni extract, the DPPH and ABTS radical scavenging activity experiments were conducted and the noni extract showed 97.0 % and 92.0 % antioxidant efficacy at 200 ㎍/mL respectively. The noni extract (50 and 100 ㎍/mL) decreased IL-6 and TNF-α in RAW 264.7 cells induced by LPS in a concentration-dependent manner. In the RT-PCR experiment involving NO production, the noni extract (50 and 100 ㎍/mL) inhibited NO production by strongly inhibiting iNOS mRNA expression, and also inhibited the elevation of MMP-1 and elastases caused by UVB irradiation by 25.0 % and 7.0 % respectively. In addition, type-1 procollagen was elevated by 20.0 % by the noni extract treatment in HaCaT cells. Conclusion: The noni extract has photoprotective ability by reducing proinflammatory mediators, elastase and MMP-1 production, and elevation of collagen synthesis. Our findings suggest that the noni extract might be a good natural substance to protect against UVB-induced premature skin aging.

• 한국자원식물학회지
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• 제31권3호
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• pp.218-227
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• 2018

In this study, we investigated the effect of the extracts from Vaccinium oldhamii on cell proliferation and the regulatory mechanisms of cyclin D1 protein level in human cancer cells. The branch extracts from Vaccinium oldhamii (VOB) showed higher inhibitor effect against the cell growth than leave extracts (VOL) and fruit extracts (VOF) in human colorectal cancer, breast cancer, prostate cancer, non-small lung cancer, pancreatic cancer and liver cancer cells. In addition, VOB decreased cyclin D1 level at both protein and mRNA level. MG132 treatment attenuated VOB-mediated cyclin D1 downregulation. A point mutation of threonine-286 to alanine attenuated cyclin D1 degradation by VOB. In addition, the inhibition of nuclear export by leptomycin B (LMB) attenuated cyclin D1 degradation by VOB. But, the treatment of PD98059 (ERK1/2 inhibitor), SB203580 (p38 inhibitor), SP600125 (JNK inhibitor), LiCl ($GSK3{\beta}$ inhibitor), LY294002 (PI3K inhibitor) or BAY 11-7082 ($I{\kappa}K$ inhibitor) did not affect VOB-induced cyclin D1 degradation. In conclusion, VOB induced cyclin D1 degradation through redistribution of cyclin D1 from the nucleus to cytoplasm via T286 phosphorylation of cyclin D1, which resulted in the inhibition of cancer cell proliferation.

• Asian Pacific Journal of Cancer Prevention
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• 제15권12호
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• pp.4951-4956
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• 2014

Purpose: To investigate IQGAP1 and IQGAP2 expression in hepatocellular carcinoma (HCC) and itsassociation with HCC clinicopathological characteristics and survival outcomes. Methods: IQGAP1 and IQGAP2 mRNA and protein were measured in HCC tissues, para-tumor tissues and normal tissues by RT-PCR and Western blotting. We further examined 150 HCC samples with adjacent para-tumor tissues and 11 normal specimens by immunohistochemistry to evaluate the correlation of IQGAP1 and IQGAP2 with clinicopathological features and prognosis. Results: IQGAP1 mRNA and protein were up-regulated while IQGAP2 mRNA and protein were down-regulated in human HCC tissues compared with para-tumor and normal liver tissues (p<0.05). IQGAP1 expression was higher in primary HCC (122/150, 81.3%) than matched adjacent tissues (30/150, 20%, p<0.001), whereas IQGAP2 was lower (31/150, 20.7% as compared to 112/150, 74.7%, P<0.001). Positive IQGAP1 expression correlated with larger tumor size (p=0.002), advanced TNM stage (p=0.002) and tumor differentiation (III and IV, p=0.034). Negative IQGAP2 expression was significantly associated with larger tumor size (p=0.009), multicentric tumor occurrence (p=0.01), advanced TNM stage (0.009) and tumor differentiation (III and IV, p=0.020). Survival analysis revealed that patients with either IQGAP1+ or IQGAP2-tumors had significantly reduced disease-free survival (p<0.001 and 0.006 respectively) and overall survival (p<0.001 for both). Multivariate analysis showed that IQGAP1/2 switch was an independent prognosis factor for disease-free survival (HR=2.824) and overall survival (HR=2.189). Conclusion: Positive IQGAP1 and negative IQGAP2 expression were closely correlated with tumor progression and could be used as adjunctive biomarkers to improve prognostication for HCC patients.

• Reproductive and Developmental Biology
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• 제33권2호
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• pp.93-98
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• 2009

In this study, bovine Dnmt1 cDNA was sequenced and detected Dnmt1 mRNA level in bovine tissues by northern blot, methylation pattern of genome by southern blot, specific localization of Dnmt1 in mouse and bovine preimplantation embryos by immunocytostaining and Dnmt1 protein level in ovary and testis by western blot. Bovine Dnmt1 cDNA sequence showed more homology with that of human than mouse and rat. The RNA level of Dnmt1 was 10 times higher expression in placenta than other tissues. This indicates that placenta was hypermethylated compared to others organs. The genomic DNA could not be cut by a specific restriction enzyme (HpaII) in placenta, lung and liver of bovine. It suggests that Dnmt1 in some somatic cells was already methylated. Dnmt1, which has the antibody epitope 1316~1616, was distributed in nucleus and cytoplasm including the stage of pronuclear stage and maturation of oocyte and gradually weaken to blastocyst stage compare to negative. In addition, Dnmt1 was strongly expressed in tetraploid embryo and cloned 8-cell than IVF 8-cell. An aberrant pattern of DNA methylation in cloned embryo may be abnormal development of fetus, embryonic lethality and placenta dysfunction. The somatic specific band (190kDa) was appeared in ovary and testis, but oocyte specific band (175kDa) was not. Further investigations are necessary to understand the complex links between the methyltransferases and the transcriptional activity of genes in the cloned bovine tissues.

• 한국한의학연구원논문집
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• 제11권2호
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• pp.97-111
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• 2005

This study was performed to evaluate anti-thrombotic activities and anti-inflammatory effects of ChungyeolHaedogHwaeoTang water extract(CHHT). The results were summarized as follows. In experiment of anti-thrombotic effect; 1. CHHT inhibited human platelet aggregation induced by ADP and epinephrine as compared with the control group, and inhibited pulmonary embolism induced by collagen and epinephrine (inhibitory rate is 37.5%). 2. CHHT increased platelet number significantly, and also CHHT shortened PT and APTT significantly as compared with the control group in thrombus model induced by dextran. In experiment of anti-inflammatory effect; 3. CHHT inhibited $IL-1{\beta}$, IL-6, TNF-${\alpha}$, COX-2 and NOS-II mRNA expression as compared with the control group in a concentration-dependent degree, and inhibited NO production significantly at 50, 100 ${\mu}g/ml$, and also inhibited ROS production in a concentration-dependent degree as compared with the control of group in RAW 264.7 cell line. 4. CHHT inhibited $IL-1{\beta}$, IL-6 and TNF-${\alpha}$ production significantly in serum of acute inflammation-induced mice, and decreased $IL-1{\beta}$, IL-6 and TNF-${\alpha}$ production in spleen tissue, and also decreased $IL-1{\beta}$, and IL-6 production in liver tissue, but increased TNF-${\alpha}$ production in liver tissue of acute inflammation-induced Balb/c mice. 5. CHHT increased survival rate from the 3rd day in ICR mice with lethal endotoxemia induced by LPS. These results suggest that CHHT can be useful in treating diverse female diseases caused by thrombosis and inflammation such as menstrual pain, menstrual disorder, leukorrhea, vaginitis, cervicitis, pelvic inflammatory disease and so on.