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도기헌,Min Sung Kim,김재호,임병용,이원석,김치대,배순식 생화학분자생물학회 2009 Experimental and molecular medicine Vol.41 No.8
Angiotensin II (AngII) is a crucial hormone that affectsvasoconstriction and exerts hypertrophic effects onvascular smooth muscle cells. Here, we showed thatphosphatidylinositol 3-kinase-dependent calciummobilization plays pivotal roles in AngII-induced vascularconstriction. Stimulation of rat aortic vascularsmooth muscle cell (RASMC)-embedded collagen gelwith AngII rapidly induced contraction. AngII-inducedcollagen gel contraction was blocked by pretreatmentwith a phosphatidylinositol 3-kinase (PI3K) inhibitor(LY294002) whereas ERK inhibitor (PD98059) was noteffective. AngII-induced collagen gel contraction wassignificantly blocked by extracellular calcium depletionby EGTA or by nifedipine which is an L-type calciumchannel blocker. In addition, AngII-induced calciummobilization was also blocked by nifedipine andEGTA, whereas intracellular calcium store-depletionby thapsigargin was not effective. Finally, pretreatmentof rat aortic ring with LY294002 and nifedipine significantlyreduced AngII-induced constriction. Giventhese results, we suggest that PI3K-dependent activationof L-type calcium channels might be involved inAngII-induced vascular constriction.
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Lysophosphatidic acid induces cell migration through the selective activation of Akt1
김은경,윤성지,도기헌,김민성,조몽,서동수,김치대,김재호,Morris J. Birnbaum,배순식 생화학분자생물학회 2008 Experimental and molecular medicine Vol.40 No.4
Akt plays pivotal roles in many physiological responses including growth, proliferation, survival, metabolism, and migration. In the current studies, we have evaluated the isoform-specific role of akt in lysophosphatidic acid (LPA)-induced cell migration. Ascites from ovarian cancer patients (AOCP) induced mouse embryo fibroblast (MEF) cell migration in a dose-dependent manner. On the other hand, ascites from liver cirrhosis patients (ALCP) did not induce MEF cell migration. AOCP-induced MEF cell migration was completely blocked by pre-treatment of cells with LPA receptor antagonist, Ki16425. Both LPA- and AOCP-induced MEF cell migration was completely attenuated by PI3K inhibitor, LY294002. Furthermore, cells lacking Akt1 displayed defect in LPA-induced cell migration. Re-expression of Akt1 in DKO (Akt1-/-Akt2-/-) cells restored LPA-induced cell migration, whereas re-expression of Akt2 in DKO cells could not restore the LPA-induced cell migration. Finally, Akt1 was selectively phosphorylated by LPA and AOCP stimulation. These results suggest that LPA is a major factor responsible for AOCP-induced cell migration and signaling specificity of Akt1 may dictate LPA-induced cell migration.
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중점관리기준에 기초한 국내생산 당귀의 산지 수확 후 아플라톡신의 안전성 평가 연구
최혜진,안태진,안영섭,박충범,김주일,박성환,양현,도기헌,문유석 한국약용작물학회 2011 한국약용작물학회지 Vol.19 No.1
HACCP methodology was applied in the post-harvest processing and storage of domestic medicinal produces. Particularly in terms of mold and mycotoxin contamination, candidate critical control points (CCP) in the conventional practice in Korean farms were selected and monitored by comparing with on the standard guided processing and storage. When each processing of Angelicae Gigantis Radix were assessed for their safety, the drying steps such as the sun drying or the thermal drying depending on each farm made differences in mold contamination. Moreover, the storage conditions before or after the processing were another critical determinant in the fungal contamination. In other words, storage under 4˚C rather than at room temperature was favorable for reducing mold growth in the harvested crops. Occurrence rate of Aflatoxin B1 (AFB1) in Angelicae Gigantis Radix were 12.8%, but amount of AFB1 in all the collected samples were below 10 ppb regulatory limit allowed in Korea. However, for a few samples of Angelicae Gigantis Radix, still relatively high levels of total amount of the major aflatoxins (aflatoxin B1 + B2 + G1 + G2) were observed around 0.18~49.94 ppb, which is not regulated presently in Korea. It thus can be suggested that post-harvest processing and storage of Korean medicinal crops need further investigation and monitoring to establish the Good Agricultural Practice (GAP), particularly to minimize microbial risk including mold and mycotoxin contamination under the changing climate. Additionally, it is also warranted for new enacting of regulatory limits for total aflatoxins in the medicinal crops.
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염증성 장질환 모델 및 크론병 환자에서의 점막상피 HuR 단백질의 변화 분석
최혜진(Hye Jin Choi),박재홍(Jae-Hong Park),박지연(Jiyeon Park),김주일(Juil Kim),박성환(Seong-Hwan Park),오창규(Chang Gyu Oh),도기헌(Kee Hun Do),송보경(Bo Gyoung Song),이승준(Seung Joon Lee),문유석(Yuseok Moon) 한국생명과학회 2014 생명과학회지 Vol.24 No.12
염증성 장질환은 점막의 만성적 궤양과 염증을 동반하는 면역질환으로 알려져 있으며, 특히 TNFα와 같은 염증성 사이토카인은 주요한 생물학적 치료의 표적으로 이용되고 있다. 염증성 사이토카인의 유전자발현에서 전사물의 안정화는 매우 중요한 조절과정이며, 특히 본 연구에서는 이 안정화에 핵심적인 단백질인 HuR의 발현과 조직분포에 대하여 동물모델과 환자의 조직에서 분석하였다. DSS를 처리함으로 유도되는 장염증 동물 모델에서 HuR 단백질의 발현량이 높았음을 확인했고, 점막의 상피조직 및 선조직 상피세포에서 상대적인 발현이 증대되었다. 또한 단백질의 활성측면에서 세포질로 이동된 HuR 단백질의 양도 상대적으로 증가하였다. 공간분포적으로 보면 DSS에 의한 화학적 점막자극에 의하여 초기에는 villi 하부에서의 발현정도가 상대적으로 villus 말단에 비하여 높게 유지되었다. 크론병 환자의 생검을 통하여 정상부위와 병변부위에서 HuR 단백질을 비교분석 하였다. 크론병 환자들의 병변에서는 지속적으로 HuR의 발현이 증대되어 있음을 확인했으며, 동물조직과 유사하게 병변부위의 장관상피세포 및 선 상피에서 주로 발현양이 높았다. 이러한 결과는 염증성 장질환에서의 HuR 단백질이 초기염증성 인자의 발현에 중요한 역할이 예상되며, 구체적인 분자기전의 규명도 향후 기대된다. 이를 근간으로 하여 염증성 장질환의 진단과 치료의 표적개발에서 유용하게 응용하고자 한다. Inflammatory bowel disease is an immune disorder associated with chronic mucosal inflammation and severe ulceration in the gastrointestinal tract. Antibodies against proinflammatory cytokines, including TNFα, are currently used as promising therapeutic agents against the disease. Stabilization of the transcript is a crucial post-transcriptional process in the expression of proinflammatory cytokines. In the present study, we assessed the expression and histological distribution of the HuR protein, an important transcript stabilizer, in tissues from experimental animals and patients with Crohn’s disease. The total and cytosolic levels of the HuR protein were enhanced in the intestinal epithelia from dextran sodium sulfate (DSS)-treated mice compared to those in control tissues from normal mice. Moreover, the expression of HuR was very high only in the mucosal and glandular epithelium, and the relative localization of the protein was sequestered in the lower parts of the villus during the DSS insult. The expression of HuR was significantly higher in mucosal lesions than in normal-looking areas. Consistent with the data from the animal model, the expression of HuR was confined to the mucosal and glandular epithelium. These results suggest that HuR may contribute to the post-transcriptional regulation of proinflammatory genes during early mucosal insults. More mechanistic investigations are warranted to determine the potential use of HuR as a predictive biomarker or a promising target against IBD.
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