Furosemide에 의한 소디움 운반체 발현의 변화

오윤규 ( Yoon Kyu Oh ),나기영 ( Ki Young Na ),이재욱 ( Jay Wook Lee ),장혜련 ( Hye Ryun Chang ),박영선 ( Young Sun Park ),박정환 ( Jung Hwan Park ),주권욱 ( Kwon Wook Joo ),김근호 ( Gheun Ho Kim ),이정상 ( Jung Sang Lee ),한진석 ( 대한신장학회 2003 Kidney Research and Clinical Practice Vol.22 No.2

배경 : 임상에서 흔히 사용하는 이뇨제 furosemide는 비후상행각에서 Na+-K+-2CI- cotranspoter (NKCC2)를 억제하여 NaCl 재흡수를 차단하여 이뇨작용을 나타낸다. Furosemide를 장기간 투여하면 내성과 대사성 알카리증의 부작용이 발생할 수 있는데, 이는 집합관에 도달하는 소디움 증가와 관련 있을 가능성이 있다. 방법 : 저자들은 furosemide의 내성이나 부작용이 집합관 상피 소디움 통로 (ENaC) 단백발현의 변화와 관련이 있는지를 확인하고자, Sprague-Dawley rat에서 farosemide (12 mg/day)을 7일간 지속적 피하 주입한 후 반정량적 immunoblotting과 면역조직화학법을 이용하여 NKCC2, Na +-CL- cotransporter (NCC), ENaC 단백의 발현을 관찰하였다. 실험기간 동안 수분과 전해질 용액 (0.8% NaCl & 0.1% KCl)을 자유롭게 섭취하도록 하여 체액 감소를 방지하였다. 결과 : 부형약 (vehicle)을 투여한 대조군에 비하여, furosemide를 투여한 군에서 각각 요량과 요 소디움 배설이 증가하였으나, 체중, 혈청 알도스테론치 및 크레아티닌 청소율은 차이가 없었다. Furosemid 투여 후 NKCC2 단백은 피질 (151±10 vs. 100±10%, p<0.05)과 외수질 (122±5 vs. 100±3%, p<0.01)에서 증가해 있었다. ENaC 단백은 세 가지 subunit 모두 furosemide 투여 후 대조군에 비하여 피질 (α:187±25 vs. 100±22%, p<0.05; β:155±8 vs. 100±15%, p<0.05; γ:168±16 vs. 100±9%, p<0.05)과 외수질 (α:171±27 vs. 100±17%, p<0.05; β :986±91 vs. 100±33%, p<0.01; γ :242±24 vs. 100±22%, p<0.01)에서 증 가하였다. 면역조직화학법에서도 furosemide를 투여한 군의 집합관 주세포에서 ENaC β-subunit가 더 강하게 염색되었다. 결론 : 이상에서 장기간 furosemide 투여시 집합관 ENaC 발현이 증가하며, 이러한 원위부네프론의 적응 반응이 이뇨제 내성을 유발하는데 기여할 것으로 생각한다. Background : Furosemide inhibit NaCl absorption in the thick ascending limb and produce an increase in distal delivery of Na+. We carried out semiquantitative immunoblotting and immunohistochemistry of rat kidneys to investigate whether chronic furosemide infusion is associated with compensatory increases in the abundance of Na+ transporters in distal nephron. Methods : Osmotic minipumps were implanted into Sprague-Dawley rats to deliver 12 mg/day of furosemide(n=6) with simultaneous administration of 0.8% NaCl and 0.1% KCl in drinking water for 7days. Results : Compared with vehicle infused controls, urine volume and urine sodium amount were increased. However, there were no differences in body weight, serum aldosterone, and creatinine clearance. The abundance of Na+-K+-2CI - cotransporter after furosemide infusion was increased in cortex (151±10 vs. 100±10%, p<0.05) and outer medulla (122±5 vs. 100±3%, p<0.01). In furosemide infusion group, the abundance of all three subunits of epithelial sodium channel (ENaC) was increased both in cortex (α:187±25 vs. 100±17%, p<0.05; β:155±8 vs. 100±15%, p<0.05; γ :168±16 vs. 100±9%, p<0.05) and outer medulla (α:171±27 vs. 100±17%, p<0.05; β :986±91 vs. 100±33%, p<0.01; γ :242±24 vs. 100±22%, p<0.01). Consistent with these results, ENaC β-subunit immunohistochemistry showed a remarkable increase in immunoreactivity in the principal cells of collecting ducts with furosemide treatment. Conclusion : These increases in the abundance of ENaC protein may account for the generation of diuretic tolerance.

제약업종 부산물 및 화장품 제조업 폐수처리오니 처리토양에 대한 유기화합물 및 Bioassay 분석 평가

임동규,이상범,이승환,남재작,나영은,권장식,권순익,소규호 한국환경농학회 2004 한국환경농학회지 Vol.23 No.4

본 연구는 퇴비원료로 지정된 제약업종 부산물(공정오니) 및 화장품 제조업 폐수처리오니를 퇴비로서 활용 가능성을 판단하기 위하여 기존의 일반성분이나 중금속 성분 분석 이외 유기성 화합물과 기타 여러 생물검정법 등을 활용하여 시용한 오니에 의한 토양 내 비료의 피해를 밝혀내고 이들의 평가방법을 확립하고자 포장시험을 수행하였다. 오니의 처리에 따른 HEM의 함량은 제약업종 오니2(PS2)와 화장품 오니(CS)처리가 각각 0.51, 1.10 mg/kg로 가장 높았고, PAHs의 함량은 제약업종 오니2(PS2) 처리에서 3,406.8 μg/kg로 가장 높았다. 토양에 서식하는 미소동물의 밀도변화는 생육 중기(7월 23일) 및 수확기(10월 1일) 모두 제약오니2 및 돈분 처리구에서 가장 높았으며, 기타 처리구는 차이가 없이 아주 낮았다. 오니의 처리에 따른 세균 및 사상균의 균수는 제약업종 오니2처리에서 각각 736, 909 cfu/g로 가장 많았고, 화장품 오니도 각각 440, 236 cfu/g으로 다른 처리에 비해 많은 경향을 보였다. 제약오니 및 화장품오니 처리시 우점세균은 무비, 돈분처리에 비해 일정한 경향이 없었으나 3요소 처리보다 다양한 세균이 검출되었다. 오니의 처리에 따른 우점사상균은 무처리에 비해 제약업종 오니3와 화장품 오니처리에서 형태적 특징이 다른 콜로니가 검출되었으나 다른 처리는 비슷한 경향으로 종류 간에 큰 차이는 보이지 않았다. Microcosm test를 통하여 오니 처리 후 3개월이 경과한 토양의 오염정도를 평가한 결과는 제약업종 오니3과 화장품 오니처리는 약간 영향을 받아서 80~90% 생존하였으나, 제약업종 오니1처리는 처리 2주(14일) 이후에는 급격히 생존율이 떨어져서 4주 이후에는 10%만 생존하였다. 그러나 6개월이 경과한 토양에서는 지렁이의 생존율은 제약업종 오니1처리만 약간 영향을 받은 것으로 보였으며 다른 처리들은 전혀 영향을 받지 않았던 것으로 조사되었다. 유기성 오니의 퇴비원료로 활용은 비료관리법의 비료공정 규격 중 퇴비의 비고란에 "퇴비의 원료로 사용 가능한 물질과 사용 불가능한 물질'(별표1)에서 "희비의 원료 중 사전 분석검토 후 사용 가능한 원료에 대한 지정요령'에는 유기물과 중금속(8성분) 함량(건물증)과 제조공정 등을 검토하여 지정하고 있으나, 지정된 원료가 과연 회비원료로 적합한지 잘 알 수 없으며 또한 앞으로 현재의 퇴비원료 규정을 변경할 필요가 있을 경우를 대비하여 희비원료의 적합성 여부를 판별할 수 있는 방법의 개발이 필요하다. 따라서 퇴비원료로 이미 지정('02. 12. 31)된 제약오니 및 화장품 오니를 과량으로 토양에 시용한 후 유해 유기화합물, 미소동물, 미생물 및 생물학적(지렁이) 유해성 검정방법의 도입 가능성을 평가하기 위하여 고추를 재배한 포장에서 비료의 피해시험을 실시한 결과 유해 유기화합물과 생물학적(지렁이) 유해성 검정방법은 앞으로 연구를 통해서 보완할 경우 상당히 활용 가능성이 있는 좋은 평가방법인 것으로 생각된다. This study was conducted to assessment organic compound and bioassay (density of inhabited animal, fluctuation of predominant fungi, and survival ratio of earthworm) for finding damage on red pepper by heavily amount application of sludges in soil, which was treated with 3 pharmaceutical byproducts and a cosmetic industry wastewater sludge as raw materials of compost, and for establishing estimation method. HEM contents in the soil treated with pharmaceutical byproducts sludge2 (PS2) and cosmetic sludge (CS) were 0.51, 1.10 mg/kg respectively. PAHs content of PS2 treatment in the soil was 3406.8 ug/kg on July 8. In abundance of soil faunas, the pharmaceutical byproducts sludge2 treatment was the most highest. The next was decreased in the order of pig manure (PM) and the cosmetic sludge treatment. However the other pharmaceutical sludge treatments were remarkably reduced populations of soil inhabited animals. In upland soil treated with organic sludges, the numbers of bacteria and fungi of the pharmaceutical sludge treatment were 736, 909 cfu/g and those of the cosmetic sludge treatment were 440, 236 cfu/g, respectively. The pharmaceutical sludge treatments and the cosmetic sludge treatment in identification of predominant bacteria were not any tendency to compare with non fertilizer and pig manure treatments, but they had diverse bacteria than NPK treatment. In microcosm tests, the survival of the tiger earthworm in five soil samples was hardly affected against the soil of PSI (20%) after three months treated in the upland. But after six months, survival of PSI was 80%. At present, raw material of compost was authorized by contents of organic matter, heavy metal (8 elements), and product processing according to "The specified gist on possible materials of using after analysis and investigation among raw materials of compost", however, for preparing to change regulation of raw material of compost and for considering to possibility of application, this study was conducted to investigate toxic organic compound and bioassay methods using inhabited animal, fungi, and earthworm without current regulation.

혈중 감마지티피 상승의 임상적 의의

신주연,김용규,박신구,이지나,김환철,임종한,홍윤철 大韓産業醫學會 2005 대한직업환경의학회지 Vol.17 No.1

목적: 혈중 감마지티피는 주로 알코올 섭취와 간세포 손상의 지표로서 이용되어져 왔으나 최근에는 혈중 감마지티피가 체내의 산소성 스트레스를 민감하게 반영하는 지표이며, 혈중 감미지티피를 이용하여 만성질환의 발생을 조기에 예측할 수 있다는 주장들이 있다. 이에 본 연구에서는 일반 인구 집단을 대상으로 한 건강검진 자료를 분석하여 혈중 감마지티피의 상승에 영향을 미치는 요인들을 분석하고 혈중 감마지티피와 산소성 스트레스와의 관련성 및 임상적인 이용 가능성을 알아보고자 한다. 방법: 2001년부터 2003년까지 한 대학병원 산업의학과에서 건강검진을 실시한 15,546명을 대상으로 하였고 신체계측, 혈액검사 그리고 설문지의 정보를 분석하였다. 총 15,546명 중 남성은 9,660명 이었고 여성은 5,886명 이었다. 연령, 성별, 체질량지수, 흡연, 음주, 운동량 비타민 복용 여부에 따른 혈중 감마지티피의 평균을 분석하였고, 성별에 따른 다변량 분석을 실시하였다. 다변량 분석에는 연령, 체질량지수, 음주빈도, 흡연량, 혈색소, 이완기혈압, 공복시혈당, 총콜레스테롤, ALT를 포함하였다. 결과: 연구 대상자들의 혈중 감마지티피의 평균은 남성에서 평균(±2SD)이 47.51(±111.62)이었고, 여성에서는 19.09(±42.02) 으로서, 대체적으로 정상 범위에 있다고 보여 지나 개인차가 상당히 크다는 사실을 알 수 있었다. 다변량 분석의 결과에서는 혈중 감마지티피의 상승에 영향을 미치는 요인이 성별에 따라 다소 달랐으나 남녀 모두에게 연령, 음주빈도, 흡연량, 이완기혈압, 공복시혈당, 총콜레스테롤, ALT가 유의한 영향이 있음을 확인할 수 있었다. 그리고 다변량 로지스틱회귀분석의 결과에서 '임상적 비정상'을 예측함에 있어 혈중 감마지티피가 진단적 유용성이 있음을 확인하였다. 결론: 긴존에 일반적으로 알려져 있는 것처럼 혈중 감마지티피를 단순히 알코올 섭취와 간세포 손상의 지표로서만 볼 수 없으며, 성별, 연령, 흡연, 고지혈증, 당뇨, 고혈압 등 많은 다른 요인들에 의해서 영향을 받으며, 혈중 감마지티피를 산소성 스트레스를 반영하는 생물학적 지표로 해석할 수도 있다. 따라서 혈중 감마지티피의 상승을 임상적으로 해석할 때는 (비록 정상인 범위에 있다하더라도), 알코올 섭취나 간세포 손상 뿐 아니라 다른 여러 요인들을 함께 고려해야 한다. 더욱이 혈중 감미지티피의 상승을 만성질환 즉, 고혈압, 당뇨, 고지혈증 등의 발생을 조기에 발견할 수 있는 지표로서 임상영역이나 건강검진시 유용하게 활용할 수 있을 것이다. Objectives: Incresed gamma-glutamyltransferase (GGT) concentration has mainly been used as a biologic marker of alcohol drinking and liver diseases. However, some recent reports have suggested that serum GGT concentration may be an early biomarker of oxidative stress and associated with chronic diseases like hypertension, DM, hyperlipidemia, CHD, stroke etc. In this study, we analysed the factors associated with increased serum GGT level and evaluated the clinical application of serum GGT in predicting the risk of chronic diseases. Methods: The data were collected from 15,546 periodic health examinations, 9,660 males and 5,886 females, done at one university hospital from 2001 to 2003. We analysed self-questionnaire, physical examination and laboratory data. Statistical analyses(t-test, ANOVA, Pearson's correlation analysis, multiple regression analysis and logistic regression analysis) were done by SPSS for windows 10.0. Results: Mean serum GGT levels were quite different according to age, sex, BMI, smoking and alcohol drinking (all P-value <0.001), but were not different by regular vitamin intake or not (P-value = 0.117). There was significant correlation between serum GGT level and the variables (of age, BMI, blood pressure, Hb, fasting blood sugar, total cholesterol, AST and ALT) (all P-value <0.001). In sex-specific multiple regression analysis, increased serum GGT level was significantly associated with age, alcohol drinking, smoking, blood pressure, fasting blood sugar and ALT in males (all P-value <0.001), and with age, alcohol drinking, smoking, Hb, blood pressure, total cholesterol and ALT in females (all P-value <0.05). In sex-specific logistic regression analysis, increased serum GGT level was significantly associated with risk of 'clinical abnormalities'. This risk increased about 13-fold in males and 4.6-fold in females for serum GGT level over the third quatile, compared with under the first quartile. Conclusions: Serum GGT level is increased not only by alcohol drinking or liver diseases, but also by many other factors associated with chronic diseases and behavioral factors. Thus many factors should be considered in evaluating an increase in GGT level, even when within the upper normal range. Serum GGT level may be a predictor for the early development of chronic diseases.

붕어의 발육 및 생식에 미치는 비소의 영향

남성숙,이철우,류지성,박응로,남규찬,류홍일,전성환,나진균,최덕일,박광식 한국환경독성학회 2001 환경독성보건학회지 Vol.16 No.2

To evaluate the arsenic effect of fish endocrine disruption, crucian carp (Carassius auratus) were treated with 0.5 mg/L and 3.0mg/L of As₂O₃ for 14 days and gonadal development was examined by histological analysis. In ovaries from female crucian carps exposed to 3.0mg/L, immature follicles and atretic follicles were appeared while various normal developmental stages of oocytes were shown in control group. Body weight and Gonadosomatic Index (GSI) of treated carp were also decreased compared to control group. However, no other significant histological changes in liver or kidney were shown in this exposure scheme, This means that reproductive organs are more sensitive than other organs and arsenic may exert endocrine disrupting effect through inhibiting the development of reproductive organ in fish.

Isolation and Characterization of Trophoblast Stem Cells-like Cells Derived from Human Term Placenta

Na Kyu-Hwan,Shin Kyung-Seon,Choi Jong-Ho,Cha Dong-Hyun,Kim Gi-Jin 한국발생생물학회 2010 발생과 생식 Vol.14 No.3

The trophectoderm is one of the earliest cell types to differentiate in the forming placenta. It is an important for the initial implantation and placentation during pregnancy. Trophoblast stem cells (TBSCs) develop from the blastocyst and are maintained by signals emanating from the inner cell mass. However, several limitations including rarity and difficulty in isolation of trophoblast stem cells derived from blastocyst still exist. To establish a model for trophoblast differentiation, we isolated TBSCs from human term placenta (38 weeks) and characterized. Cell cycle was analyzed by measuring DNA content by FACS analysis and phenotype of TBSCs was characterized by RT-PCR and FACS analysis. TBSCs have expressed various markers such as self-renewal markers (Nanog, Sox2), three germ layer markers (hNF68, alpha-cardiac actin, hAFP), trophoblast specific markers (CDX-2, CK7, HLA-G), and TERT gene. In FACS analysis, TBSCs isolated from term placenta showed that the majority of cells expressed CD13, CD44, CD90, CD95, CD105, HLA-ABC, cytokeratin 7, and HLA-G. Testing for CD31, CD34, CD45, CD71, vimentin and HLA-DR were negative. TBSCs were shown to decrease the growth rate when cultured in conditioned medium without FGF4/heparin as well as the morphology was changed to a characteristic giant cell with a large cytoplasm and nucleus. In invasion assay, TBSCs isolated from term placenta showed invasion activities in in vivo using nude mice and in vitro Matrigel system. Taken together, these results support that an isolation potential of TBSCs from term placenta as well as a good source for understanding of the infertility mechanism.

Altered expression of norepinephrine transporter and norepinephrine in human placenta cause pre-eclampsia through regulated trophoblast invasion

Na, Kyu-Hwan,Choi, Jong Ho,Kim, Chun-Hyung,Kim, Kwang-Soo,Kim, Gi Jin The Korean Society for Reproductive Medicine 2013 Clinical and Experimental Reproductive Medicine Vol.40 No.1

Objective: We investigated the norepinephrine transporter (NET) expression in normal and pre-eclamptic placentas and analyzed the invasion activity of trophoblastic cells based on norepinephrine (NE)-NET regulation. Methods: NET and NE expression levels were examined by western blot and enzyme-linked immunosorbent assay, respectively. Trophoblast invasion activity, depending on NE-NET regulation, was determined by NET-small interfering RNA (siRNA) and NET transfection into the human extravillous trophoblast cells with or without NE treatment and invasion rates were analyzed by zymography and an invasion assay. Results: NET mRNA was expressed at a low level in pre-eclamptic placentas compared with normal placentas and NE concentration in maternal plasma increased significantly in pre-eclamptic women compared to normal pregnant women (p<0.05). NET gene upregulation and NE treatment stimulated trophoblast cell invasion up to 2.5-fold (p<0.05) by stimulating matrix metalloproteinase-9 activity via the phosphoinositol-3-kinase/AKT signaling pathway, whereas NET-siRNA with NE treatment reduced invasion rates. Conclusion: NET expression is reduced by inadequate regulation of NE levels during placental development. This suggests that a complementary balance between NET and NE regulates trophoblast cell invasion activities during placental development.

Poster Session : Model Organisms and Disease ; Inhibition of nitric oxide and tumor necrosis factor-alpha by Moutan Cortex in activated mouse peritoneal macrophages

( Hwan Suck Chung ),( Moon Kyu Kang ),( Chong Woon Cho ),( Jae Yoon Kim ),( Na Youn Lee ),( Chong Heong Park ),( Dong Woo Kim ),( Joong Hwan Oh ),( Hong Yeoul Kim ),( Min Kyu Shin ),( Moo Chang Hong ) 한국생화학분자생물학회 (구 한국생화학회) 2007 생화학분자생물학회 춘계학술발표논문집 Vol.2007 No.-

Electron beam sensitivity and DNA damage in imidacloprid-resistant and -susceptible strains of Aphis gossypii

Seung-Hwan Yun,Sang-Eun Park,Soon-Kyu Lee,Hyun-Na Koo,Gil-Hah Kim 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.10

The effect of electron beam irradiation on development and reproduction of imidacloprid-resistant (R strain) and -susceptible (S strain) Aphis gossypii were compared. Nymphs and adults of R and S strain were irradiated at target doses of 50, 100, 150, and 200 Gy. When nymphs were irradiated, emergence was not affected at all target dose, but number of F1 nymph was perfectly inhibited at 150 Gy in both strains. When irradiated to the adults, longevity slightly decrease at 150 Gy and above. Fecundity was strongly decreased at 100 Gy, but was not completely inhibited even at 200 Gy. Emergence of F1 nymph was decreased at 100 Gy and completely inhibited at 200 Gy. However, there were no significant differences on development and reproduction of R and S strain. We also conducted the comet assay immediately after irradiation and over the following 10 day period. Severe DNA fragmentation in A. gossypii cells was observed just after irradiation and the damage was repaired during the post-irradiation period in a time-dependent manner. These results suggest that electron beam irradiation induced abnormal development, reproduction, and DNA damage in A. gossypii, but there were no significant differences between R and S strain.

광반응성 천연고분자의 생체재료로써의 응용

나하나 ( Ha Na Na ),허윤 ( Yoon Heo ),김광일 ( Kwang Il Kim ),정규환 ( Kyu Hwan Chung ),손태일 ( Tea Il Son ) 한국키틴키토산학회 2010 한국키틴키토산학회지 Vol.15 No.2

본 총설에서는 광반응성을 갖는 천연 고분자를 생체재료로 응용하기 위하여 생체재료로써의 요구조건과 특성을 살펴보았다. 생체재료로 사용하는 천연고분자 중 단백질을 기초로 하는 고분자와 다당류, 단당류 유도체들로 나눌 수 있다. 이러한 천연고분자는 독성이 적어서 생체 적합성이 우수하고, 생분해가 가능하며, 합성 고분자 물질에 비해 경제성이 있는 장점을 가진다. 이러한 천연고분자에 광반응성기를 도입하면 천연고분자의 표면이 개질 될 수 있기 때문에 기존의 천연고분자를 생체재료로 사용하게 될 때의 세포조직에 대한 단점을 보완할 수 있을 것이라 사료된다. Artificial organs were used in medical circle all over the world, because biomaterial was also developed as applied materials. Biomaterial is artificial materials which are exposed by intermittent or continuing contact with surrounding tissue for function of tissues in vivo. Using photo-reactive natural polymer introduced above characteristics as biomaterial and functional groups caused photo reaction, in this paper, we describe the applications of photo-reactive natural polymer as a biomaterial which can be used various fields.

Survival of Salmonella enterica and Listeria monocytogenes in Chicken and Pig Manure Compost

Kyu-Seok Jung,Sung-Gi Heu,Eun-Jung Roh,Min-Ha Kim,Hyun-Ji Gil,Na-Young Choi,Dong-Hwan Lee,Jeong-A Lim,Jae-Gee Ryu,Kye-Hoon Kim 한국토양비료학회 2013 한국토양비료학회지 Vol.46 No.6

Livestock manure is a valuable source of nutrients and organic matter for plant. Thus, livestock manure compost is commonly used fertilizer in organic vegetable and fruit production in many countries. However, contaminated or inadequate manure compost can give negative effect to soil microorganisms. This study was conducted to investigate the survival difference of Salmonella enterica and Listeria monocytogenes in chicken and pig manure compost under the selected environmental conditions. Commercially available manure compost (pig, chicken) was inoculated with S. enterica and L. monocytogenes. Manure compost was incubated at 25℃ and consistent moisture content. Samples had been collected during 200 days depending on the given conditions. S. enterica survived for 130 days in pig manure compost and over 200 days in chicken manure compost, respectively. L. monocytogenes persisted for 120 days in pig manure compost and over 200 days in chicken manure compost, respectively. It is noted that the number of S. enterica and L. monocytogenes gradually decreased over time. The results indicate that S. enterica survived longer than L. monocytogenes in manure compost at 25℃. S. enterica and L. monocytogenes survived longer in chicken manure compost than in pig manure compost. Increased knowledge of pathogen behavior in agricultural environments is a valuable part of future work on improving risk evaluations and, in a longer perspective, in providing data for guidelines regarding safe handling of pathogen-contaminated manure compost and soil.