정상 및 무칼륨 식이 백서 신장에서 Na⁺-K⁺-ATPase Subunit Isoform mRNA 발현에 관한 연구

안규윤(Kyu Youn Ahn),김석채(Sug Chae Kim),문 범(Bum Moon),김경근(Kyung Keun Kim),김백윤(Baik Yoon Kim) 대한해부학회 1998 Anatomy & Cell Biology Vol.31 No.3

저칼륨혈증은 여러 조직에서 Na+-K+-ATPase 유전자 발현의 정도를 변화시키며 또한 신장의 속질바깥층집합세관에서는 Na+-K+-ATPase 활성과 α1과 β1 subunit 단백을 증가시키지만 겉질집합세관에서는 유의한 변화가 없다는 것은 잘 알려져 있다. 그러나 집합세관을 위시한 다른 콩팥단위 부위에서의 Na+-K+-ATPase subunit isoform들의 적응반응에 관한 연구는 없는 실정이다. 이에 저자들은 정상 및 무칼륨 식이 백서 신장에서 Na+-K+-ATPase subunit isoform mRNA 발현의 상대적인 양과 분포를 구명하기 위해 Northern 분석과 in situ hybridization 조직화학을 실시하였다. Northern 분석 결과, 대조군의 Na+-K+-ATPase subunit isoform mRNA들은 속질바깥층, 겉질 및 속질속층 순으로 발현되었 으나 실험군에서는 속질바깥층, 속질속층 및 겉질 순으로 발현되었다. α1 mRNA 양은 α2, α3 보다 훨씬 많았고, 각 isoform에 대한 mRNA의 양은 속질속층에서 대조군 보다 실험군에서 2~3배 이상 증가하였으나 겉질과 속질바깥층에서는 두 군간에 유의한 차이가 없었다. In situ hybridization 조직화학 결과에서 각 isoform에 대한 mRNA의 분포는 대조군의 토리쪽세관의 S3 부위, 겉질상행후각, 속질상행후각, 먼쪽곱슬세관 및 전 집합세관에서 관찰되었다. 실험군 백서에서 α1, α3 및 β1 isoform의 반응정도는 속질상행후각, 겉질상행후각, 먼쪽곱슬세관 및 겉질집합세관에서 감소하였으나 속질바깥층집합세관 내층 상부와 속질속층집합세관 상부에서는 현저히 증가하였다. α2 isoform은 속질상행후각에서는 감소하였으나 겉질상행후각, 먼쪽곱슬세관 및 겉질집합세관에서 약간 증가하였고 속질바깥층집합세관 내층 상부와 속질속층집합세관 상부에서는 현저히 증가하였다. 이상의 소견으로 저칼륨혈증에 의해 Na+-K+-ATPase subunit isoform 유전자들의 발현은 속질바깥층집합세관 내층 상부와 속질속층집합세관 상부에서 증가되며, 이들 부위에서 유전자들이 K 이온 보존에 관여하고 있음을 암시해 준다. Chronic hypokalemia alters Na+-K+-ATPase gene expression in several tissues. While it is established that Na+- K+-ATPase activity and α1 and β1 subunit protein levels increase during K depletion in the outer medullary collecting duct (OMCD) and do not significantly change in the cortical collecting duct (CCD), little is known about the adaptive responses of the other isoforms in these other nephron segments. Accordingly, this study was performed to characterize the relative levels of expression and cellular distribution of mRNAs encoding the Na+-K+-ATPase subunit isoforms in normal and K-deprived (2 weeks) rats using the Northern analysis and in situ hybridization (ISH). Isoform specific 32Plabeled cDNA (for Northerns) or digoxigenin labeled cRNA (for ISH) probes were used. In normal rats, the order of expression amounts of all isoforms mRNAs from highest was outer medulla > cortex > inner medulla , and that of K-deprived rats was outer medulla > inner medulla > cortex. α1 mRNA levels were much greater than those of α2 or α3 in cortex, outer and inner medulla. mRNA levels for all isoforms were 2~3 folds greater in inner medulla of K-deprived rats compared to controls. In contrasts, the levels of all isoforms mRNAs in cortex and outer medulla were comparable between the two gruops. By ISH, mRNAs for all isoforms were observed in the S3 segment of proximal tubule, the cortical thick ascending limb (CTAL), medullary thick ascending limb (MTAL), distal convoluted tubule (DCT), connecting tuble (CNT), and the entire collecting duct. Both groups exhibited comparable cellular patterns of labeling, but the signal intensity of K-deprived rats was much greater in the proximal portion of the inner stripe of outer medullary collecting duct (OMCDi) and proximal portion of the inner medullary collecting duct (IMCD), and less in the MTAL compared to controls. The signal intensity of α1, α3, and β1 isoforms was less in the CTAL, DCT, and CCD of K-deprived rats, but α2 isoform was slightly increased. These results suggest that chronic hypokalemia enhances expression of Na+-K+-ATPase subunit isoforms in the proximal portion of OMCDi and proximal IMCD, but not other nephron segments, and that these isoforms may participate in potassium conservation by these segments during potassium deprivation.

인태아(人胎兒) 활액막세포(滑液膜細胞)의 전자현미경적(電子顯微鏡的) 연구(硏究)

윤재룡,전철홍,안규윤,Yoon, Jae-Rhyong,Chun, Cheol-Hong,Ahn, Kyu-Youn 한국현미경학회 1994 Applied microscopy Vol.24 No.1

The development of synovial membrane from knee joint was studied by electron microscope in human fetuses ranging from 20mm to 260mm crown rump length (40days to 30weeks of gestational age). At 40mm fetus, developing synovial tissue was observed in homogenous interzone as a vascular mesenchyme around the periphery. The primitive joint space was appeared after the intermediate layer of the interzone in direct contact with chondrogenic layer at 60mm fetus. Differentiation of the synovial membrane coincided with clarification of the joint cauity. When dilatation of the synovial cavity occurred, the two types of synovial cells were well endowed with rough endoplasmic reticulum. At 100mm fetus, type A cells with a markedly attenuated cytoplasm were found as well as those cells which contained pinocytotic vesicles and vacuoles. By 150-200mm fetuses a majority of the intimal cells were type B. These cells were characterized by abundant rough endoplasmic reticulum and well developed Golgi complex. In contrast, A-type cell had numerous filopodia, pinocytotic vesicles lysosomes, and large vacuoles containing amorphous material. At 260mm fetus, the intimal cells were well developed and plentiful. The most marked difference between the synovial membrane of full-term fetus and adult was the large amount of collagen in the latter. During fetal period, the B-cells were most numerous cell type in the intimal cells. The B-cells were clearly distinguishable from the A-cells by their content of extensive rough endoplasmic reticulum and well developed Golgi complex.

흰쥐 눈물샘에서 Carbonic Anhydrase 발현에 관한 면역조직화학적 연구

안 민(Min Ahn),이송은(Song Eun Lee),남광일(Kwang Il Nam),정채용(Chaeyong Jung),이승원(Seung-won Lee),안규윤(Kyu Youn Ahn),배춘상(Choon Sang Bae),김백윤(Baik Yoon Kim),박성식(Sung Sik Park) 대한해부학회 2008 Anatomy & Cell Biology Vol.41 No.1

흰쥐 안구 밖 눈물샘에서 carbonic anhydrase (CA) 동종효소의 발현 여부를 관찰하고자 흰쥐 눈물샘을 대상으로 CA I, II, IV 및 IX 동종효소에 대한 항체를 사용하여 면역조직 화학 염색을 시행하였고, 이들 조직에서 각각의 CA 동종효 소 단백 발현을 Western blotting 분석을 시행하여 확인하였다. Western blotting 분석 결과 CA II와 IX가 가장 강하게, CA I이 중등도로, CA IV가 가장 약하게 발현되었다. H-E 염색에서 안구 밖 눈물샘은 대롱꽈리샘으로 많은 소엽과 사이막으로 구성되어 있었다. 소엽에는 장액샘꽈리와 사이관 그리고 소집합관들이 관찰되었다. 면역조직화학 염색에서 CA I 반응은 샘꽈리세포에서는 음성이었으나 사이관세포와 소집합관세포에서는 양성 반응을 보였다. CA II 반응은 샘꽈리세포의 경우 샘꽈리에 따라 다양한 양성 반응을 보였으며 양성 부위는 샘꽈리세포의 핵상부 세포질이었다. 사이관과 소집합관에서는 대부분 음성이었으나 일부 사이관과 소집합관은 약한 반응을 보였다. CA IV 반응은 샘꽈리세포에서는 음성이었으나 사이관 세포와 소집합관세포는 강한 양성을 보였다. CA IX 반응은 대부분의 샘꽈리세포에서는 음성이었으나 극히 일부 샘세포에서는 바닥세포막에서 양성 반응을 보였다. 사이관과 소집합관세포에서는 양성 반응을 보였다. 이상의 관찰로 흰쥐 안구 밖 눈물샘에서 샘꽈리세포와 도관세포에 분포하는 CA 동종효소가 다름을 확인하였으며, 눈물샘에서의 전해질 대사는 샘꽈리세포는 주로 CA II에 의해 이루어지고 도관계는 주로 CA I, IV 및 IX에 의해 이루어질 것으로 추측되었다. There are several carbonic anhydrase (CA) isozymes, which differ in their kinetic properties, tissue distribution, and subcellular localization. In this study, the distribution of CA isozymes I, II, IV, and IX was investigated in the rat exorbital lacrimal gland using Western blotting analysis and immunohistochemical staining. In the Western blotting analysis of the rat lacrimal gland, CA II and CA IX were expressed abundantly and CA IV was expressed weakly. Hematoxylin-eosin staining of the exorbital lacrimal gland showed a multilobular tubuloacinar gland composed of acinar and ductal cells. Immunohistochemical reaction revealed no CAI staining in acinar cells and positive staining in intercalated and small duct cells. CA II reactivity was detected in the supranuclear cytoplasm of acinar cells and appeared to vary between acini. The intercalated and collecting duct cells showed weak or no immunoreactivity for CA II. CA IV was detected in the intercalated and collecting duct cells but not at the acinar cells. CA IX was detected in the intercalated and collecting duct cells, and in only a few acinar cells. These results demonstrate the differential distribution of CA isoenzymes in the exorbital lacrimal gland of the rat and suggest that CA II is related mainly to the electrolyte metabolism of tears in the acinar cells and that CAs I, IV, and IX are related to the electrolyte metabolism of tears in the duct cells.

토끼에서 αvβ5 인테그린 항체의 결막하주사에 의한 각막신생혈관 억제효과

윤경철,임성규,오한진,안규윤,김경근,Kyung-Chul Yoon,Seong-Kyu Im,Han-Jin Oh,Kyu-Youn Ahn,Kyung-Keun Kim 대한안과학회 2005 대한안과학회지 Vol.46 No.11

Purpose: To investigate the efficacy of a subconjunctival injection of αvβ5 integrin antibody on corneal angiogenesis induced by chemical epithelial denudation in a rabbit eye model. Methods: One week after debridement by heptanol, rabbits were treated with a subconjunctival injection of αvβ5 integrin antibody or control immunoglobulin G weekly for 2 weeks. Rabbits that did not receive injection after debridement served as the untreated group. The percentage of neovascularized corneal area was calculated by biomicroscopy, and the sectioned area and number of new vessels were calculated by histological examinations. Results: At 7 days after the first injection, αvβ5 integrin antibody-treated eyes had 9.5% (P=0.02) and 6.8% (P=0.03) less neovascularized corneal area than vector-treated eyes and untreated eyes, respectively. At 7 days after the second injection, αvβ5 integrin antibody-treated eyes had 21.1% (P=0.02) and 18.3% (P=0.02) less neovascularized corneal area than vector-treated eyes and untreated eyes, respectively. Light microscopic examination showed a smaller neovascularized corneal area and a reduced number of new vessels in the αv β5 integrin antibody-treated eyes compared to the control eyes. Conclusions: Subconjunctival injection of αvβ5 integrin antibody effectively reduces experimental corneal neovascularization induced by chemical injury, and could be used as a corneal angiogenesis inhibitor in the future.

저칼륨혈증 흰쥐 신장에서 Nrf2 전사유전자의 발현 증가

임진섭 ( Jin Seob Lim ),안규윤 ( Kyu Youn Ahn ) 대한신장학회 2011 Kidney Research and Clinical Practice Vol.30 No.3

Purpose: The purpose of this study was to analyze the expression and localization of Nrf2 mRNA in rats according to the changes of K-diet. Methods: Nrf2 gene was isolated using DNA chip microassay. Northern and Western blot analysis and in situ hybridization (ISH) were performed. Results: Northern analysis of normal rat demonstrated that Nrf2 mRNA was abundantly expressed in stomach, moderately in testis, kidney, distal colon, duodenum, and adrenal gland, and weakly in brain, heart, spleen, salivary gland, liver, and lung. In the kidney of K-restricted groups, Nrf2 mRNA and protein expressions were gradually increased in K-restricted kidney. By ISH, hybridization signal for Nrf2 gene of normal group was prominent in the S3 segment of proximal tubule, distal tubule, and cortical collecting duct, and weak in outer and inner medullary collecting duct. In K-restricted groups, the localization of hybridization signal was the same as in normal group. Signal intensity of K-restricted groups was markedly increased in outer and inner medullary collecting ducts compared with normal group. But, that of the distal tubule and cortical collecting duct was decreased. mRNA for Nrf2 gene of normal group was detected in the cells of the basal portion of intestinal gland of distal colon and stomach, spermatogonia and spermatocytes of seminiferous tubule of testis, small lymphocytes of germinal center of spleen, and adrenal medulla cells of adrenal gland. Conclusion: These results suggest that expression of Nrf2 is different in various tissues and increased expression of Nrf2 gene in outer and inner medullary collecting ducts of hypokalemic kidney could regulate the ion transporter genes by these segments.

각막신생혈관에 대한 버테포르핀을 이용한 광역학치료의 동물실험

나현주,윤경철,임욱빈,안규윤,서만성,Hyeon-Ju Nah,M,N,Kyung-Chul Yoon,Wook-Bin Im,Kyu-Youn Ahn,Man-Seong Seo 대한안과학회 2005 대한안과학회지 Vol.46 No.4

Purpose: To determine the efficacy of photodynamic therapy (PDT) with verteporfin (Visudyne?, Norvatis Ophthalmics AG, Hettingen, Switzerland), a benzoporphyrin derivative, in the treatment of corneal neo- vascularization (CN) in a rabbit eye model. Methods: CN was induced by placing instrastromal sutures in the cornea. Two weeks after suturing, verteporfin was administrated intravenously and 1 hour later, the right eye (treated group) was exposed to a laser with a 689 nm wavelength, and the left eye was used as the control. The changes in CN were analyzed using biomicroscopy and optical microscopy in twelve rabbits. Results: The mean percentages of the neovascular area in the control and treated groups were 96.4±1.9% and 90.3±3.5% (P=0.009) at three days after the PDT, 88.6±4.6% and 71.6±6.2% (P<0.001) at one week, and 76.8±4.4%와 43.6±15.1% (P<0.001) at two weeks, respectively. Optical microscopy showed significant differences between the control and treated group in terms of the area and number of CN (P<0.05). Conclusions: PDT with verteporfin is a safe and effective procedure for regressing CN. However, a further study will be necessary.

저칼륨혈증 흰쥐 신장에서 Nrf2와 p-Nrf2 단백발현의 변화

조혜정(Hye Jung Cho),안규윤(Kyu Youn Ahn) 대한체질인류학회 2015 해부·생물인류학 (Anat Biol Anthropol) Vol.28 No.1

K<SUP>+</SUP>평형조절에 이온통로나 펌프 유전자 이외에도 다양한 유전자가 관여한다. 저칼륨혈증 신장에서 NF-E2-related factor 2 (Nrf2) mRNA의 분포와 Nrf2 전사유전자가 이온수송체를 조절한다는 것은 밝혀져 있지만 Nrf2와 phosphorylated-Nrf2 (p-Nrf2) 단백의 분포는 밝혀지지 않았다. 이에 본 연구는 칼륨제한 식이에 따른 흰쥐 신장 내 Nrf2와 p-Nrf2의 발현 및 분포의 변화를 면역조직화학적 방법으로 관찰하였다. 면역조직화학적 소견에서 Nrf2 단백의 면역반응성은 토리쪽곱슬세관과 토리쪽곧은세관에서는 높게 발현되었고, 겉질 굵은오름부분에서는 중등도의 발현을 보였으며 바깥속질 굵은오름부분과 겉질집합관 및 바깥속질집합관에서는 낮게 발현되었다. 칼륨제한 식이군의 Nrf2의 발현부위는 정상 식이군과 비교해 차이가 없었으나 면역 반응성은 칼륨제한 식이 3주군의 토리쪽곱슬세관과 토리쪽곧은세관에서 최대발현을 보였다. 정상 식이군 p-Nrf2 단백의 면역반응성은 겉질 굵은오름부분, 겉질집합관 및 사구체내피세포의 핵에서는 높게 발현되었고, 먼쪽곱슬세관과 바깥속질집합관에서는 중등도의 발현을 보였으며, 토리쪽곱슬세관과 바깥속질 굵은오름부분의 핵에서는 낮게 발현되었다. 칼륨제한 식이군의 p-Nrf2의 발현부위는 정상 식이군과 비교해 유사하였으며 면역반응성은 칼륨제한 식이 2주군과 3주군의 바깥속질집합관의 핵에서 현저히 증가하였으며 그 외 겉질집합관, 먼쪽곱슬세관, 겉질 굵은오름부분 및 바깥속질 굵은오름부분에서는 중등도의 증가를 보였으며, 겉질 토리쪽곱슬세관에서는 정상 식이군과 유사하였다. 이상의 결과로 저칼륨혈증에서 신장 Nrf2와 p-Nrf2의 단백발현 양상은 상호 간에 차이를 나타냈으나 면역반응성은 칼륨제한 식이가 길어질수록 증가함을 보여주었다. 또한 이온수송체 단백의 발현부위와 유사하여 이들 유전자의 조절뿐만 아니라 세포 내 신호전달에 있어 중요한 역할에 관여할 것임을 시사해 주었다. Potassium (K) balance is regulated not only by ion channels and ion transporters, but also by various genes including NF-E2-related factor 2 (Nrf2). Although mRNA distribution and role of Nrf2 has been studied in hypokalemic kidney, the distribution of Nrf2 and phosphorylated-Nrf2 (p-Nrf2) proteins are not known. The present study was planned to examine the alteration of expression and distribution of Nrf2 and p-Nrf2 protein in the kidney of normal and K-depleted rats using immunohistochemistry. In normal rat kidneys, Nrf2 was highly expressed in the proximal convoluted tubule and proximal straight tubule, moderately in cortical thick ascending limb, and weakly in cortical collecting duct, outer medullary thick ascending limb, and outer medullary collecting duct. In K-depleted groups, the pattern of cellular labeling of Nrf2 protein was identical to that of normal group, but the signal intensity was prominently increased in proximal convoluted tubule and proximal straight tubule especially in rats at K-free diet 3 weeks. In normal rat kidneys, p-Nrf2 was highly expressed in nucleus of cortical thick ascending limb, cortical collecting duct, and glomerular endothelial cell, moderately in distal convoluted tubule and outer medullary collecting duct, and weakly in proximal convoluted tubules and outer medullary thick ascending limb. In K-depleted groups, the pattern of cellular labeling of p-Nrf2 protein was similar to that of normal group, but signal intensity was significantly increased in the nucleus of outer medullary collecting duct from of K-free diet 2 and 3 weeks groups. These results suggest that Nrf2 and p-Nrf2 expression was gradually increased in K-depleted groups of kidney, but Nrf2 and p-Nrf2 expression patterns were not exactly matched. In addition, it is suggested that enhanced expression of Nrf2 and p-Nrf2 in hypokalemic condition may affect the regulation of ion channels and ion transporters and subsequent intracellular signal transduction.

임신 흰쥐 신장에서 Aquaporin-1, 2, 3 단백발현의 변화

조혜정(Hye Jung Cho),이창배(Chang Bae Lee),안규윤(Kyu Youn Ahn) 대한해부학회 2008 Anatomy & Cell Biology Vol.41 No.4

임신은 모체의 장기에 해부?생리학적으로 많은 변화를 초래하고 신장에서는 세포외액 및 혈장량 증가로 나트륨 및 수분저류가 일어난다. 수분대사의 항상성 및 수분대사장애는 aquaporin (AQP)의 양적인 변화와 관련이 있다고 알려져 있지만 임신시 AQP 발현에 관한 연구는 AQP-2뿐이다. 이에 본 연구는 흰쥐 신장에서 임신기간에 따른 AQP-1, 2, 3 단백발현 및 분포의 변화를 Westhern 분석 및 면역조직화학적 방법으로 관찰하였다. Western 분석소견에서 AQP-1, 3 단백은 임신 17.5일에 최대발현을 보였고, AQP-2 단백은 임신 19.5일에 최대발현을 보였다. 면역조직화학적 소견에서 AQP-1 단백은 비임신 신장의 겉질의 토리쪽세관과 바깥속질 및 속속질의 Henle 고리 내림세관의 첨부세포막에서 관찰되었다. 각 임신시기의 면역반응 부위는 비임신과 차이가 없었으나 면역반응성은 임신 10.5일과 임신 12.5일에서 증가하기 시작하여 임신 17.5일에 최대발현을 보였다. AQP-2 단백은 비임신 신장의 겉질집합관과 바깥속질집합관 및 속속질집합관 주세포의 첨부세포막과 세포질에서 관찰되었다. 각 임신시기의 면역반응 부위는 비임신과 차이가 없었으나 면역반응성은 임신 10.5일과 12.5일에는 중등도의 발현을 보이다가 임신 17.5일에 증가하여 임신 19.5일에 최대발현을 보였다. AQP-3 단백은 겉질집합관과 바깥속질집합관 및 속속질집합관 주세포의 기저외세포막에서 관찰되었다. 각 임신시기의 면역반응 부위는 비임신과 차이가 없었으나 면역반응성은 AQP-2 단백발현과 마찬가지로 임신 10.5일부터 증가하여 임신 19.5일에 최대발현을 보였다. 이상의 소견으로 임신 중 특히 말기에서 세포외액 및 수분증가는 AQP-1, 2, 3 단백발현의 조절에 의해서 이루어질 것으로 생각되었다. The pregnancy causes the marked change in maternal renal hemodynamic and volume homeostasis. During pregnancy, renal sodium and water retention result in an expansion of extracellular fluid and plamsma volume. Although many studies suggested that water balance or water balance disorder was associated with regulation of Aquaporin (AQP) expression, the studies were only limited to AQP-2 expression during the pregnancy. The present study was to examine altered expression and distribution of AQP-1, 2, and 3 proteins in the kidneys of non-pregnant (NP) and pregnant rats using Westhern blot analysis and immunohistochemistry. Pregnant Sprague-Dawley rats were evaluated on various time sets: days 10.5 (P10.5), 12.5 (P12.5), 17.5 (P17.5), and 19.5 (P19.5). In Westhern blot analysis, expression of AQP-1, 2 was peaked at P17.5 and AQP-3 at 19.5. Immunoreactivity of AQP-1 of NP rat was detected in the apical membranes of proximal tubules and thin limb of Henle loop. In pregnant rats, the pattern of cellular labeling of AQP-1 protein was identical to NP rat, but signal intensity was continuously increased from P10.5 and peaked at P17.5. In NP rat, immunoreactivity of AQP-2 was the most prominent in apical region and moderate in cytoplasm of the principal cells of entire collecting duct. In pregnant rats, the pattern of cellular labeling of AQP-2 protein was identical to NP rat, but signal intensity was moderately expressed in P10.5 and P12.5 and most prominent signal was observed in P19.5. In NP rat, immunoreactivity of AQP-3 was most prominent in the bosolateral plasma membrane of principal cells of entire collecting duct. In pregnant rats, the pattern of cellular labeling of AQP-3 protein was identical to NP rat, but signal intensity was continuously increased from P10.5 to P17.5 and peaked at P19.5. These results suggest that the expansion of extracellular fluid volume and water retention are regulated by AQP-1, 2, and 3 during the pregnancy, especially at late stage.

사람 창자에서 Carbonic Anhydrase 동위효소 발현에 관한 연구

이동진(Dong Jin Lee),남광일(Kwang Il Nam),이승원(Seung Won Lee),안규윤(Kyu Youn Ahn),배춘상(Choon Sang Bae),김백윤(Baik Yoon Kim),박성식(Sung Sik Park) 대한해부학회 2004 Anatomy & Cell Biology Vol.37 No.4

사람 소화관 상피에서 carbonic anhydrase (CA)의 발현 여부를 관찰하고자 샘창자와 주름창자를 대상으로 CAI, CAII, CAIV, CAIX 동위효소에 대한 항체를 사용하여 면역조직화학 염색을 시행하였고, 이들 조직에서 각각의 CA 동위효소 단백 발현을 Western blot을 시행하여 확인하였다. Western blot에서 CAI, IX는 샘창자와 주름창자에서 모두 강하게 발현되었다. CAII는 샘창자와 주름창자의 점막조직에서 강하게 발현되었고, CAIV는 샘창자와 주름창자의 점막조직에서 약하게 발현되었다. 샘창자에서 면역조직화학 염색 결과 CAI은 음성이었다. CAII는 융모원주세포와 점막밑샘세포 및 도관세포에서 양성이었으나 창자샘에서는 음성이었다. CAIV는 융모상피와 창자샘에서는 음성이었으나 점막밑샘에서는 도관세포가 양성반응을 보였다. CAIX는 융모상피는 음성이었으나 창자샘은 양성이었고 점막밑샘에서는 극히 일부의 샘세포에서 양성이었다. 주름창자에서 면역조직화학 염색 결과 CAI과 II는 상피 원주세포와 술잔세포에서 양성이었다. CAIV와 IX는 상피의 원주세포에서만 양성이었다. 이상의 관찰로 샘창자에서 표면상피는 CAII가, 창자샘은 CAIX이, 점막밑샘에서 분비세포는 CAII가, 도관세포는 CAII와 IV가 가장 강하게 발현되고, 주름창자에서 표면상피는 CAI, II, IV, IX 모두 발현되나 창자샘은 모두 음성임을 알 수 있었다. 이상의 결과는 샘창자 점막밑샘은 샘창자에서의 산-염기 균형 조절에 CAII와 IV를 통해 관여할 것임을 시사하였다. The distribution of carbonic anhydrase (CA) isozymes I, II, IV, and IX was investigated in the human duodenum and colon using Western blotting analysis and immunohistochemical staining. A Western blotting analysis revealed an abundant expression of CAI and IX in the duodenum and colon. The expression of CAII and IV was detected in mucosa of duodenum and colon. The degree of expression, however, showed regional difference. The expression of CAII was strong in the duodenum and colon, and that of CAIV was weak in the duodenum and colon. Immunohistochemical staining of duodenum revealed no staining for CAI. CAII was detected at the columnar cells of surface epithelium and secretory cells and ductal cells of Brunner’s gland. CAIV was detected at the ductal cells of Brunner’s gland. CAIX was detected at the cells of intestinal gland and rare cells of the Brunner’s gland. Immunohistochemical staining of colon showed a positive reaction of CAI and II at the columnar and goblet cells of surface epithelium, and CAIV and IX at the columnar cells of surface epithelium. These results demonstrate the differential distribution of CA isozymes in duodenum and colon, and suggest that Brunner’s gland may contribute the control of acid-base balance in the duodenal lumen by secreting bicarbonate ion catalyzed by CAII and IV.

수분, 전해질 및 산 염기 이상 : 흰쥐 신장에서 kNBC1의 조절인자 연구

이창배 ( Chang Bae Lee ),조혜정 ( Hye Jung Cho ),이지연 ( Ji Youn Lee ),이송은 ( Song Eun Lee ),안규윤 ( Kyu Youn Ahn ) 대한신장학회 2008 춘계학술대회 초록집 Vol.28 No.1