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Chemical inhibitors destabilize HuR binding to the AU-rich element of TNF-α mRNA
Min-Ju Chae,Hye Youn Sung,Eun-Hye Kim,Mira Lee,Hojoong Kwak,Chong Hak Chae,Sunwoo Kim,Woong-Yang Park 생화학분자생물학회 2009 Experimental and molecular medicine Vol.41 No.11
Hu protein R (HuR) binds to the AU-rich element (ARE) in the 3'UTR to stabilize TNF-α mRNA. Here, we identified chemical inhibitors of the interaction between HuR and the ARE of TNF-α mRNA using RNA electrophoretic mobility gel shift assay (EMSA) and filter binding assay. Of 179 chemicals screened, we identified three with a half-maximal inhibitory concentration (IC50) below 10 μM. The IC50 of quercetin, b-40, and b-41 were 1.4, 0.38, and 6.21 μM, respectively, for binding of HuR protein to TNF-α mRNA. Quercetin and b-40 did not inhibit binding of tristetraprolin to the ARE of TNF-α mRNA. When LPS-treated RAW264.7 cells were treated with quercetin and b-40, we observed decreased stability of TNF-α mRNA and decreased levels of secreted TNF-α. From these results, we could find inhibitors for the TNF-α mRNA stability, which might be used advantageously for both the study for post-transcriptional regulation and the discovery of new anti-inflammation drugs. Hu protein R (HuR) binds to the AU-rich element (ARE) in the 3'UTR to stabilize TNF-α mRNA. Here, we identified chemical inhibitors of the interaction between HuR and the ARE of TNF-α mRNA using RNA electrophoretic mobility gel shift assay (EMSA) and filter binding assay. Of 179 chemicals screened, we identified three with a half-maximal inhibitory concentration (IC50) below 10 μM. The IC50 of quercetin, b-40, and b-41 were 1.4, 0.38, and 6.21 μM, respectively, for binding of HuR protein to TNF-α mRNA. Quercetin and b-40 did not inhibit binding of tristetraprolin to the ARE of TNF-α mRNA. When LPS-treated RAW264.7 cells were treated with quercetin and b-40, we observed decreased stability of TNF-α mRNA and decreased levels of secreted TNF-α. From these results, we could find inhibitors for the TNF-α mRNA stability, which might be used advantageously for both the study for post-transcriptional regulation and the discovery of new anti-inflammation drugs.
저산소성 허혈성 뇌손상을 유발시킨 어린 백서에서 Myeloperoxidase 측정 검사
양혜정(Hae Joung Yang),피대훈(Dae Hun Pee),임지혜(Ji Hye Lim),최병민(Byung Min Choi),이기형(Kee Hyoung Lee),은백린(Baik-Lin Eun) 대한소아신경학회 2002 대한소아신경학회지 Vol.10 No.2
목 적 : 예전에는 뇌경색 부위에서 발견되는 백혈구를 단지 뇌손상에 뒤따르는 생리적인 반응으로만 생각하였으나 최근 10년 사이 재관류 손상에서도 백혈구에 의한 염증 작용이 중요한 역할을 한다고 알려지면서 이들의 역할에 대한 연구가 새롭게 이루어지고 있다. PML의 존재는 세포질내 과립에서 분비되는 MPO를 생화학적으로 측정함으로서 증명할 수 있는데 저자들은 미성숙 뇌에서 저산소성 허혈증에 의한 뇌손상이 발생할 때 백혈구가 침윤되는 과정을 MPO 측정 검사를 통하여 관찰하였고, P와 L-selectin 억제제인 Fucoidin을 투여한 후 MPO의 변화를 확인하였다. 방 법 : 생후 7일된 어린 흰쥐를 사용하여 우측총 경동맥을 전기 응고시켜 자르고 8% 산소에 노출시켰다. 저산소 노출 후 회복 시간에 따라 동물을 희생시키고 뇌를 추출하여 뇌조직 1 g당 10mL의 20 mM potassium Phosphate buffer(pH 7.4)를 첨가하여 50초간 분쇄하였다. 각각의 균질회된 조직 샘플들은 4℃에서 20분 동안 원심 분리하여 상층액을 제거한 후, 침전물은 처음 조직량에 따라 조직 1 g당 10 mL의 0.5% cetylditrimethylethyl ammonium bromide(wt/vol)가 첨가된 50 mM potassium phosphate buffer(pH 6.0)를 첨가하였다. Vortex를 이용하여 buffer와 침전물을 잘 섞고 60℃수조에서 120분 동안 방치하였다. 그 후 각 조직 샘플들을 4 watts에서 음파처리하고 4℃에서 15분 동안 원심 분리를 시행하여 상층액을 새로운 tube에 옮겼다. MPO 측정검사를 위한 MPO reaction buffer는 50 mM potassium phosphate buffer(pH 6.0) 100 mL에 0.53 mM odianisidine edihydrochlorde와 0.0005% HO를 섞어 만들었다. MPO reaction solution 2.9 mL에 각각의 MPO sample 0.1 mL을 첨가하여 460 nm의 파장에서 5분 동안 MPO에 다른 흡광도 차이를 관찰하였다. 결 과 : 조직내 백혈구 침윤의 지표로 측정한 MPO 활성도는 대조군에서는 미미하였으나 총 경동백을 절단한 우측 대뇌군에서는 허혈 및 저산소증 유발 후 8시간째부터 현저히 증가되기 시작하여 24시간 경과 후 발현이 가장 높았으며, Fucoidin 50 mg/kg으로 전처치한 약물 실험군에서는 MPO 활성도가 현저히 감소하였다. 결 론 : 백혈구는 미성숙 뇌의 저산소성 허혈증에 의한 뇌손상에서 중요한 매개체 역할을 하며, 백혈구의 생화학적 활성도를 나타내는 MPO 활성도는 저산소성 허혈성 뇌손상의 정도를 측정하는 지표로 이용될 수 있다고 생각된다. Purpose : Neutrophils found around an infarcted area in the brain was once considered as only the physiologic response following the brain injury, but recent studies have shown that inflammatory responses by neutrophils play an important role in the reperfusion injury. The presence of ploymorphonuclear leukocytes(PML) is proven by biochemical assay of myeloperoxidase(MPO) Secreted in the cytoplasmic granules. We observed the process of PML infiltration on hypoxic-ischemic brain injury of immature rats by the assay of MPO activity and changes of the MPO activity after the administration of fucoidin, inhibitor of P- and L-selectin. Methods : We used a well characterized model of the brains of 7 day-old-rats, which had unilateral hypoxic and ischemic injuries(HI). Those injuries were induced by unilateral carotid artery ligation followed by timed exposure to hypoxic inspiratory gas mixture(8% O). MPO activity was measured in the brain tissue homogenates of HI rats(n=18) at 0, 2, 8, 24 and 48 hrs and in rats that received fucoidin immediately before and again after hypoxia(50 mg/kg, n=6) at 8 and 24 hrs. Controls(n=2) were rats with neither hypoxia nor ischemia. The brain samples were homogenized in 20 mM potassium phosphate buffer(pH 7.4) for 50 secs. The homogenate was centrifuged at 14,000 g at 4℃ for 15 mins and the supernatant was discarded. The tissue was pulverized, weighed, and suspended in 1 mL of 50 mM potassium phosphate buffer solution(pH 6.0) contatining 0.5% cetylditrimethylammonium bromide(wt/vol). The tissue was sonicated and centrifuged at 10,000 g for 15 mins. 200 µL of the supernatant was mixed with 1 mL of 50 mM potassium phosphate buffer solution(pH 6.0) containing 10 μL of 1.325 mM o-dianisidine hydrochloride and 170 µL of 3% hydrogen peroxide(vol/vol). Changes in absorbance at 460 nm were measured for 5 mins by using microplate reader. One unit of MPO activity was defined as that degrading 1 µmol peroxide/min at 25℃, and the result was expressed as units of MPO/100 gm tissue. Results : In HI rats, MPO activity increased at 2 hrs after HI and peaked at 24 hrs in the right hemisphere. In rats with fucoidin treatment immediately before and again after hypoxia, the MPO activity significantly decreased in both hemispheres compared with HI rats(P<0.05). MPO activity in the tissue of control rats was insignificant. Conclusion : The dynamic changes of the MPO activity suggest the important role of PMN on hypoxic-ischemic brain injuries in immature rats. MPO activity could be used as an index of the severity of injuries of hypoxic-ischemic brains.
NAM, SANG HAE,CHOI, SANG YOUK,CHOI, DONG MYEONG,YANG, MIN SUK 경상대학교 유전공학연구소 1990 遺傳工學硏究所報 Vol.9 No.-
In order to improve genetic traits of tomato(Lycopersicon esculentum) by introduction of foreign genes into existing genomes of the cell, estabilishment of reproducible cultural methods from protoplasts to regenerated plants is a preconditional procedure. In early stage of culture, the regeneration of plant from protoplasts was found to be influenced by BA(benzyl adenine) concentration and cultural period. The most abundant regenerative structure was obtained when 0.2㎎/l IAA and 2㎎/l BA were added into MS medium. We investigated changes of protein and amino acid contents and patterns throughout regengration periods. A prominent polypeptides(24 KD) and amino acids were significantly increased as the cultural period extended without producing new proteins in regeneration producedure. The ratio of aspartic acid and methionine contents of regenerative structure was rapidly increased, while that of leucine and histidine contents was slightly decreased.
다문화가족 청소년의 학교생활적응을 위한 다문화가족지원조례 개정 방안: 경기도와 충청북도 및 안산시와 충주시 비교를 중심으로
민기채 ( Min Kichae ),김혜원 ( Kim Hyewon ),반소영 ( Ban Soyoung ),사유림 ( Sa Yurim ),양유라 ( Yang Yura ),이호진 ( Lee Hojin ),조요셉 ( Jo Yosep ) 한국보건사회연구원 2021 保健社會硏究 Vol.41 No.1
이 연구의 목적은 경기도와 충청북도 및 안산시와 충주시의 다문화가족지원조례 비교를 통해 충주시 다문화가족지원조례 개정 방안을 도출함으로써 다문화가족 청소년의 학교생활적응을 위한 법적 기초자료를 생산하는 것이다. 이를 위해 먼저 규범적 체계인조례의 목적, 자치단체장의 책무, 정책 대상을 비교하였다. 다음으로 실효성 체계인 지원범위(학교생활 적응을 위한 교육지원, 한국어 교육, 심리·정서 지원, 차별 개선, 학교밖 청소년의 사회적응 지원, 사회·문화 적응 지원)와 서비스 공급체계 기반 구축 관련 조례를 비교하였다. 분석결과, 지방자치단체들의 조례는 표준조례에 기초하고 있어 대동소이하나, 충청북도는 경기도에 비해, 충주시는 안산시에 비해 조례의 세부조항이 마련되어 있지 않아 다문화가족 청소년을 위한 조례 내용이 구체적이지 않았다. 향후 충주시 다문화가족지원조례 개정 방향으로 정책대상 및 지원범위 구체화, 자치단체장의 책무 확대, 서비스 공급체계 기반 구축을 제시하였다. 다문화가족지원조례가 구체적이고 명확하게 규정되어 있을수록 서비스 제공에 있어서 질적인 차이가 있다. 본 연구를 통해 다문화가족 청소년의 학교적응을 지원하기 위한 법적 토대가 마련되고, 지역 간조례 비교 연구가 확산되기를 기대한다. The purpose of this study is to produce legal basic data for school life adaptation of multicultural family adolescents to by deriving a plan for revising the multicultural family support ordinances in Chungju city through comparison of the ordinances in Gyeonggi-do and Chungcheongbuk-do, Ansan-si and Chungju-si. To this end, the purpose of the ordinance, the responsibilities of the local government heads, and the targets of policies, which is the normative system, were compared. Next, the scope of support (educational support for school life adaptation, Korean language education, psychological and emotional support, improvement of discrimination, support for social adaptation of out-of-school youth, support for social and cultural adaptation) and ordinances related to the establishment of a service supply system base, which is the effective system, were compared. As a result of the analysis, there are no big differences because the ordinances of local governments are based on standard ordinance. However, the ordinances for multicultural family adolescents are less detailed in Chungcheongbuk-do compared to Gyeonggi-do, and Chungju-si compared to Ansan-si. In the direction of the future revision of the Ordinance for Multicultural Family Support in Chungju, it suggested the specification of policy targets and scope of support, expansion of responsibilities of local government heads, and establishment of the service supply system. The more concretely and clearly stipulated in the Multicultural Family Support Ordinance, there is a qualitative difference in service provision. It is hoped that this study will be instrumental in providing a legal basis for school life adaptation of multicultural family adolescents and in spreading comparative studies of local ordinances.
나노에멀젼 젤로 만든 고기능성치약을 이용한 구강 연조직 창상 및 병소에 대한 치험례
채창훈(Chang-Hoon Chae),최동주(Dong-Ju Choi),심혜영(Hae-Young Shim),변은선(Eun-Sun Byun),홍순민(Soon-Min Hong),박양호(Yang-Ho Park),박준우(Jun-Woo Park) 대한구강악안면외과학회 2007 대한구강악안면외과학회지 Vol.33 No.6
It is a gel type high functional toothpaste containing vitamin C, E, propolis extract and the rest of herb with a nanoemulsion state. Vitamin C, E is known as the material with an eminent anti-oxidation effect. Propolis is known as the material with an antimicrobial and anti-inflammatory effect. We have been succeeding in making nanoemulsion of vitamin C, E and propolis through the high pressure homogenizer using stable oil and lecithin and the gel type high functional tooth paste were made from nanoemulsion of vitamin C, E and propolis. We observed the process of wound protecting effect and cure effect for a wound of soft tissue, gingival tissue and mucous membrane showing ulcer and inflammation in oral cavity after applying a gel type high functional toothpaste to patient. As a result, the wound were healed very fast and any side effects were not shown. We confirmed that a gel type high functional toothpaste with nanoemulsion of vitamin C, E and propolis extract has good effect not only for wound healing but also for treatment of ulcer-like lesion in oral cavity. So we report our cases with review of literatures.