• 동의생리병리학회지
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• 제20권4호
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• pp.957-965
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• 2006

This study was peformed to evaluate antithrombotic activities and anti-inflammatory effects of Kami-BoyangHwanoh-Tang(KBHT). The major findings were summarized as follows. In experiment of anti-thrombotic effect; KBHT 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 50 %). KBHT increased platelet number significantly and also KBHT shortened PT and APTT significantly as compared with the control group in thrombus model induced by dextran. In experiment of anti-inflammatory effect; KBHT 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 group in RAW 264.7 cell line. KBHT 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-6 and $TNF-{\alpha}$ production in liver tissue, but increased $IL-1{\beta}$ production in liver tissue of acute inflammation-induced mice. KBHT increased survival rate at 3rd day in mice with lethal endotoxemia induced by LPS. These results suggest that KBHT can be useful in treating diverse female diseases caused by thrombosis and inflammation such as endometrosis, myoma, pelvic congestion, chronic cervicitis, chronic pelvic inflammatory disease and so on.

• Biomolecules & Therapeutics
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• 제26권6호
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• pp.599-607
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• 2018

Fasiglifam (TAK-875) a G-protein coupled receptor 40 (GPR40) agonist, significantly improves hyperglycemia without hypoglycemia and weight gain, the major side effects of conventional anti-diabetics. Unfortunately, during multi-center Phase 3 clinical trials, unexpected liver toxicity resulted in premature termination of its development. Here, we investigated whether TAK-875 directly inflicts toxicity on hepatocytes and explored its underlying mechanism of toxicity. TAK-875 decreased viability of 2D and 3D cultures of HepG2, a human hepatocarcinoma cell line, in concentration-(>$50{\mu}M$) and time-dependent manners, both of which corresponded with ROS generation. An antioxidant, N-acetylcysteine, attenuated TAK-875-mediated hepatotoxicity, which confirmed the role of ROS generation. Of note, knockdown of GPR40 using siRNA abolished the hepatotoxicity of TAK-875 and attenuated ROS generation. In contrast, TAK-875 induced no cytotoxicity in fibroblasts up to $500{\mu}M$. Supporting the hepatotoxic potential of TAK-875, exposure to TAK-875 resulted in increased mortality of zebrafish larvae at$25{\mu}M$. Histopathological examination of zebrafish exposed to TAK-875 revealed severe hepatotoxicity as manifested by degenerated hypertrophic hepatocytes with cytoplasmic vacuolation and acentric nuclei, confirming that TAK-875 may induce direct hepatotoxicity and that ROS generation may be involved in a GPR40-dependent manner.

• Asian Pacific Journal of Cancer Prevention
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• 제15권18호
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• pp.7527-7532
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• 2014

Saussurea involucrata is a Mongolian medicinal plant well known for its effects in promoting blood circulation, and anti-inflammation and analgesic functions. Earlier studies reported that Saussurea involucrata has anticancer activity. The purpose of this study was to confirm the anticancer activity of an ethanol extract of Saussurea involucrata against hepatic cancer and elucidate its mechanisms of action. Hepatocellular carcinoma cells were tested in vitro for cytotoxicity, AO/EB staining for apoptotic cells, apoptotic DNA fragmentation and cell cycle distribution in response to Saussurea involucrata extract (SIE). The mRNA expression of caspase-3,-9 and Cdk2 and protein expression of caspase-3,-9, PARP, XIAP, Cdk2 and p21 were analyzed through real time PCR and Western blotting. Treatment with SIE inhibited HepG2 cell proliferation dose- and time-dependently, but SIE only exerted a modest cytotoxic effect on a viability of Chang human liver cells. Cells exposed to SIE showed typical hallmarks of apoptotic cell death. Cell cycle analysis revealed that SIE caused G1-phase arrest in HepG2 cells. In conclusion, Saussurea involucrata ethanol extract has potential cytotoxic and apoptotic effects on human hepatocellular carcinoma cells. Its mechanism of action might be associated with the inhibition of DNA synthesis, cell cycle (G1) arrest and apoptosis induction through up-regulation of the protein expressions of caspase-3,-9 a nd p21, degradation of PARP and down-regulation of the protein expression of Cdk2 and XIAP.

• BMB Reports
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• 제34권5호
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• pp.434-439
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• 2001

The effects of regular endurance exercise, or acute-exercise and rest on the levels of lipids, carnitines and carnitine palmitoyltransferase-I (CPT-I) were investigated in male Sprague-Dawley rats. The rats were exercise trained on a treadmill for 60 min per day for 60 days (long-term trained, LT), or non-trained for 59 days (NT) and exercised for 60 min on the 60th day. In NT rats, the levels of serum nonesterified carnitine (NEC), acidsoluble acylcarnitine (ASAC), and total carnitine (TONE) increased significantly during the post-exercise recovery period (PERP). In LT rats, ASAC, and TCNE, which increased right after the 60 min running session decreased to the levels of pre-exercise during the PERP. The levels of skeletal muscle ASAC in NT rats, which increased significantly by the acute-exercise, decreased to the pre-exercise levels during the PERP. However, the ASAC level in LT rats reached its peak at 4 h after running for 60 min. Liver triglyceride (TG) and total lipids (TL), which increased by the acute-exercise, decreased to the pre-exercise levels during the PERP in both NT and LT rats. CPT-I activity in NT rats increased significantly after 1 h of a 60-min exercise and slowly decreased to pre-exercise levels during the PERP. However, the CPT-I activity in LT rats, which increased significantly by the 60 min exercise, decreased slowly and reached its pre-exercise level within 8 h of the PERP. Northern blot analysis showed that the changes of CPT-I activities during the PERP coincided with changes in CPT-I mRNA levels. This study shows that both regular endurance exercise, and acute-exercise and rest, can influence differently the levels of carnitines, lipids and CPT-I in rats. The results suggest that regular endurance exercise, rather than the acute-exercise, can change effectively the distributions of carnitines, lipids and CPT-I in rats during exercise and rest.

• BMB Reports
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• 제35권6호
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• pp.623-628
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• 2002

The Th1 vs. Th2 balance is critical for the maintenance of immune homeostasis. Therefore, the genes that are selectively-regulated by the Th1 and Th2 cytokines are likely to play an important role in the Th1 and Th2 immune responses. In order to search for and identify the novel target genes that are differentially regulated by the Th1/Th2 cytokines, the human PBMC mRNAs differentially expressed upon the stimulation with IL-4 or IL-12, were screened by employing the differential display-polymerase chain reaction. Among a number of clones selected, DC21 was identified as a novel target gene that is regulated by IL-4 and IL-12. The DC21 gene expression was up-regulated either by IL-4 or IL-12, yet counter-regulated by co-treatment with IL-4 and IL-12. DC21 is a dendritic cell protein with an unknown function. The sequence analysis and conserved-domain search revealed that it has two AU-rich motifs in the 3'UTR, which is a target site for the regulation of mRNA stability by cytokines, and that it belongs to the N-acetyltransferase family. The induction of DC21 by IL-12 peaked around 8-12 h, and lasted until 24 h. LY294002 and SB203580 significantly suppressed the IL-12-induced DC21 gene expression, which implies that PI3K and p38/JNK are involved in the IL-12 signal transduction pathway that leads to the DC21 expression. Furthermore, tissue blot data indicated that DC21 is highly expressed in tissues with specialized-resident macrophages, such as the lung, liver, kidney, and placenta. Together, these data suggest a possible role for DC21 in the differentiation and maturation of dendritic cells regulated by IL-4 and IL-12.

• Asian Pacific Journal of Cancer Prevention
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• 제13권5호
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• pp.2009-2014
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• 2012

Objective: Taspine, isolated from Radix et Rhizoma Leonticis has demosntrated potential proctiective effects against cancer. Tas13D, a novel taspine derivative synthetized by structure-based drug design, have been shown to possess interesting biological and pharmacological activities. The current study was designed to evaluate its antiproliferative activity and underlying mechanisms. Methods: Antiproliferative activity of tas13D was evaluated by xenograft in athymic mice in vivo, and by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and cell migration assays with human liver cancer (SMMC-7721) cell lines in vitro. Docking between tas13D and VEGFR and EGFR was studied by with a Sybyl/Surflex module. VEGF and EGF and their receptor expression was determined by ELISA and real-time PCR methods, respectively. Results: Our present study showed that tas13D inhibited SMMC-7721 xenograft tumor growth, bound tightly with the active site of kinase domains of EGFR and VEGFR, and reduced SMMC-7721 cell proliferation (IC=34.7 ${\mu}mol/L$) and migration compared to negative controls. VEGF and EGF mRNAs were significantly reduced by tas13D treatment in a dose-dependent manner, along with VEGF and EGF production. Conclusion: The obtained results suggest that tas13D inhibits tumor growth and cell proliferation by inhibiting cell migration, downregulating mRNA expression of VEGF and EGF, and decreasing angiogenic factor production. Tas13D deserves further consideration as a chemotherapeutic agent.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.5397-5402
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• 2013

The rates of morbidity and mortality of hepatocellular carcinoma (HCC) have not lessened because of difficulty in treating tumor metastasis. Mongolian Saussurea involucrata (SIE) possesses various anticancer activities, including apoptosis and cell cycle arrest. However, detailed effects and molecular mechanisms of SIE on metastasis are unclear. Thus, the present study was undertaken to investigate antimetastatic effects on HCC cells as well as possible mechanisms. Effects of SIE on the growth, adhesion, migration, aggregation and invasion of the SK-Hep1 human HCC cell line were investigated. SIE inhibited cell growth of metastatic cells in dose- and time-dependent manners. Incubation of SK-Hep1 cells with $200-400{\mu}g/mL$ of SIE significantly inhibited cell adhesion to gelatin-coated substrate. In the migration (wound healing) and aggregation assays, SIE treated cells showed lower levels than untreated cells. Invasion assays revealed that SIE treatment inhibited cell invasion capacity of HCC cells substantially. Quantitative real time PCR showed inhibitory effects of SIE on MMP-2/-9 and MT1-MMP mRNA levels, and stimulatory effects on TIMP-1, an inhibitor of MMPs. The present study not only demonstrated that invasion and motility of cancer cells were inhibited by SIE, but also indicated that such effects were likely associated with the decrease in MMP-2/-9 expression of SK-Hep1 cells. From these results, it was suggested that SIE could be used as potential anti-tumor agent.

• 대한한방내과학회지
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• 제31권3호
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• pp.650-666
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• 2010

Objectives : In this study, the author tried to investigate whether wood vinegar produced from Morus alba (MA) significantly affects the increase in airway epithelial mucosubstances and hyperplasia of tracheal goblet cells of rats, and in vitro airway mucin secretion and PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production / gene expression from human airway epithelial cells. Materials and Methods : For the in vivo experiment, the author induced hypersecretion of airway mucus and goblet cell hyperplasia by exposure of rats to SO2 over 3 weeks. Effect of orally-administered MA over 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats and hyperplasia of goblet cells were assessed using histopathological analysis after staining the epithelial tissue with alcian blue. For the in vitro experiment, confluent RTSE cells were chased for 30 min in the presence of MA to assess the effect of MA on mucin secretion by enzyme-linked immunosorbent assay (ELISA). Also, effects of MA on PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of MA and treated with PMA (10 ng/ml), EGF (25 ng/ml) or TNF-alpha (0.2 nm) for 24 hrs, to assess both effects of MA on PMA- or EGF- or TNF-alpha-induced MUC5AC mucin production by enzyme-linked immunosorbent assay (ELISA) and gene expression by reverse transcription-polymerase chain reaction (RT-PCR). Possible cytotoxicities of MA in vitro were assessed by examining LDH release from RTSE cells and the rate of survival and proliferation of NCI-H292 cells. In vivo liver and kidney toxicities of MA were evaluated by measuring serum GOT/GPT activities and serum BUN/creatinine concentrations of rats after administering MA orally. Results : 1. MA decreased the amount of intraepithelial mucosubstances of rats exposed to sulfur dioxide inhalationally. 2. MA decreased in vitro mucin secretion from cultured RTSE cells. 3. MA significantly inhibited PMA-, EGF-, and TNF-alpha-induced MUC5AC mucin productions and the expression levels of MUC5AC mRNA from NCI-H292 cells. 4. MA did not show either in vitro or in vivo hepatic or renal toxicities. Conclusion : The results from this study suggests that MA can regulate the secretion, production and gene expression of airway mucin observed in diverse respiratory diseases accompanied by mucus hypersecretion and does not show in vivo toxicity to liver and kidney functions after oral administration. Effects of MA should be further studied using animal experimental models that simulate the diverse pathophysiology of respiratory diseases via future research.

• 한국응용과학기술학회지
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• 제38권6호
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• pp.1383-1392
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• 2021

호장근, 적하수오, 대황, 알로에 등과 같은 생약재의 주요 약리 활성 성분인 emodin은 항산화, 항균, 항염, 항암, 간보호 등에 효능이 있는 것으로 보고되었다. 본 연구에서는 emodin의 피부 질환 및 기능성 소재로서의 활용 가능성을 알아보기 위해 염증 개선과 피부장벽기능 개선 관련 활성을 확인하였다. human keratinocyte인 HaCaT 세포에 대하여 항염효과를 관찰하기 위해 cytokine억제능은 ELISA kit로, 단백질 발현은 western blot으로 확인하였다. TNF-α (10 ng/mL)/IFN-γ (10 ng/mL)로 활성화된 HaCaT 세포에서 emodin을 농도별(5, 10, 20, 40) µM로 처리한 결과 TNF-α, IL-1β 및 IL-6의 생성량은 emodin의 농도가 증가함에 따라 감소됨을 확인하였다. 염증관련 단백질인 iNOS, COX-2 발현량에 대한 실험결과에서도 emodin 20 µM 농도에서 control 대비 iNOS는 48%, COX-2는 29%가 저해됨을 확인하였다. 피부장벽기능 개선의 지표로써 filaggrin, involucrin, loricirn의 mRNA 발현 정도와 filaggrin, involucrin, loricirn의 생성량을 확인한 결과 에모딘 농도에 의존적으로 증가하는 우수한 결과가 얻어졌다. 특히 20 µM 농도에서 대조군 대비 2배의 생성량 증가를 보인 filaggrin은 천연보습인자인 NMF의 형성에 관계되는 단백질로 각질층의 보습에 중요한 역할을 하는 것으로 알려져 있다. 결론적으로 emodin의 피부 질환 및 기능성 소재로서의 활용 가능성 중 염증 개선 및 피부장벽기능 개선 소재로 활용될 수 있음을 확인하였으며 향후 추가적인 연구가 수행되면 그 활용 범위가 더 넓어질 수 있을 것으로 사료된다.

• 한국미생물·생명공학회지
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• 제32권4호
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• pp.297-306
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• 2004

열 충격단백질(Heat shock protein : HSP)은 온도 스트레스에 대하여 세포 내에서 발현되는 단백질이다. HSP의 중요 분류군의 하나가 HSP90 family 이다. 여러 종류의 포유동물과 조류에서 HSP 유전자 특성에 대한 연구가 많이 진행되었다. 본 연구에서는 넙치(Paralichthys olivaceus)로부터 제조한 넙치 뇌 cDNA 유전자 은행을 이용하여 넙치 HSP90 cDNA 유전자를 분리하여 구성 염기서열의 특성을 밝혀 내었다. 염기서열의 분석결과 넙치의 hsp90$\beta$ 유전자는 2,791 개의 뉴클레오타이드로 구성되어 있고, 726개의 아미노산 잔기가 암호화되어 있었다. 넙치 hsp90$\beta$ 유전자는 European sea bass와 96.6% zebrafish와 92.9%, Atlantic salmon와 92.0%, 그리고 사람과는 89.5%의 염기서열 상동성을 지니고 있었다. 또한 HSP90 아미노산 서열을 바탕으로 척추동물 종들과의 진화계통수를 구축하였다. 넙치 hsp90$\beta$ 유전자의 mRNA의 분포 정도를 RT-PCR를 이용하여 조사하였다. hsp90$\beta$ 유전자는 조사한 모든 조직(뇌, 간, 신장, 근육, 비장)에서 높은 수준으로 발현이 되고 있었다. 또한, 넙치 hsp90$\beta$ 단백질을 대량발현하기 위하여 대장균에서 발현을 유도하였다.