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흰쥐의 간섬유화 과정에서의 교원질의 다형성 및 양적 변동
조준승,류현모,은선진,김인산 ( Joon Seung Jo,Hyun Mo Ryoo,Sun Jin Eun,In San Kim ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.4
Macromolecular collagen components in rat liver at different stage of fibrosis induced by common bile duct ligation were studied. Animals were killed on the 3rd, 7th, 14th, 21st and 29th day after bile duct ligation and liver specimens and blood samples were obtained. Hepatic collagen was fractionated into neutral salt soluble, acid soluble, pepsin soluble and insoluble fractions, and hydroxyproline content of each fraction was determined. The collagen content, especially in insoluble fraction, was increased as the duration of biliary obstruction increased. Pepsin solubilized collagens were subjected to determination of the collagen types by SDS-polyacrylamide disc gel electrophoresis. The ratio of type I to type III collagen was decreased gradually with progression of fibrosis. These results indicate that an alteration in tissue collagen polymorphism as well as variations in the collagen solubility accompany the fibrotic process, suggesting possible pathogenetic implication.
Collagen Dynamics and Polymorphism in Rat Liver During Fibrogenesis
조준승,류현모,은선진,김인산,Jo, Joon-Seung,Ryoo, Hyun-Mo,Eun, Sun-Jin,Kim, In-San Korean Society for Biochemistry and Molecular Biol 1991 한국생화학회지 Vol.24 No.4
Macromolecular collagen components in rat liver at different stage of fibrosis induced by common bile duct ligation were studied. Animals were killed on the 3rd, 7th, 14th, 21st and 29th day after bile duct ligation and liver specimens and blood samples were obtained. Hepatic collagen was fractionated into neutral salt soluble, acid soluble, pepsin soluble and insoluble fractions, and hydroxyproline content of each fraction was determined. The collagen content, especially in insoluble fraction, was increased as the duration of biliary obstruction increased. Pepsin solubilized collagens were subjected to determination of the collagen types by SDS-polyacrylamide disc gel electrophoresis. The ratio of type I to type III collagen was decreased gradually with progression of fibrosis. These results indicate that an alteration in tissue collagen polymorphism as well as variations in the collagen solubility accompany the fibrotic process, suggesting possible pathogenetic implication. 백서의 총담관을 결찰하여 간섬유화를 유발하고, 그 진행에 따른 간조직내 교원질의 양적 및 질적 변화를 조사하였다. 실험동물의 총담관을 결찰하고 각각 3,7,14,21 그리고 28일째 되는 날에 얻은 간조직과 혈액을 시료로 사용하였다. 간조직내 교원질을 neutral soluble, acid soluble, pepsin soluble 그리고 insoluble fraction으로 분획채취 하였으며, 각 분획의 hydroxyproline 함량으로 교원질을 측정하였다. 담관폐쇄 이후 간기능의 저하가 나타났고, 기간이 길어짐에 따라 간경화의 척도인 간장과 비장의 종대 및 복수의 생성을 관찰하였다. 또한 담관 폐쇄기간이 길수록 간조직의 교원질 함량은 증가하였으며, 특히 insoluble fraction의 증가가 주를 이루었다. Pepsin soluble fraction을 SDS-PAGE한 결과, type I에 대한 type III 교원질의 상대적 비율이 정상조직의 20%에서 4주군에서는 45%까지 증가하였다. 이러한 결과는 섬유화가 진행됨에 따라 교원질의 양적인 변화 뿐 아니라 용해도의 변화 혹은 결합조직을 형성하는 교원질 구성의 변화 등 질적인 변화가 발생함을 보여주고 있으며, 섬유화의 발생기작과 관계가 있을 것으로 볼 수 있다.
허혈성 급성신부전에서 TGF-β-induced gene product βig-h3의 변화와 초기 표지자로서의 의의
최민정 ( Min Jeong Choi ),박선희 ( Sun Hee Park ),김찬덕 ( Chan Duck Kim ),김용림 ( Yong Lim Kim ),권태환 ( Tae Hwan Kwon ),김인산 ( In San Kim ),김용진 ( Yong Jin Kim ) 대한신장학회 2007 Kidney Research and Clinical Practice Vol.26 No.3
Purpose : Acute renal failure remains a potentially devastating clinical problem. This study aimed to examine whether the expression of TGF-β-induced gene product, βig-h3, is altered in ischemiareperfusion (I/R) injury and urinary excretion of βig-h3 is changed in I/R injury. Methods : I/R injury was performed by clamping both renal arteries. Daily urine output, serum creatinine and urinary TGF-β and βig-h3 were measured after I/R injury. Also, the renal expression of βig-h3 by western blotting and immunohistochemistry were investigated. In the second step, urinary βig-h3 was measured at 4, 10, 16, and 24 hours after I/R injury to investigate whether it could be used as an early and sensitive marker for detecting I/R injury. Results : Urinary βig-h3 was significantly elevated at 24 hours and maintained higher than the controls until 2 days after I/R injury. In contrast, western blotting did not reveal any changes of βig-h3 expression. Immunohistochemistry showed that labeling of βig-h3 was seen at the basement membranes of proximal tubule cells mainly located at the medullary ray (S3 segment) in both groups. Following I/injury, the labeling was also seen in the basement membrane of injured or regenerated proximal tubular epithelial cells. Within 24 hours, urinary βig-h3 was significantly increased at 4 hours after I/R injury. Importantly, the urinary appearance of βig-h3 preceded that of N-acetyl-β-D-glucosaminidase. Conclusion : These results suggest that endogenous renal βig-h3 may serve to promote tissue regeneration in I/R injury and urinary βig-h3 could be used as an early and sensitive marker demonstrating I/R injury.
류마티스관절염에서 TGF-β-inducible Gene-h3 (βig-h3)에 의한 활막세포 부착기전
조현정 ( Hyung Jung Cho ),박재용 ( Jae Yong Park ),경희수 ( Hee Soo Kyung ),김인산 ( In San Kim ),강영모 ( Young Mo Kang ),남언정 ( Eon Jeong Nam ),송은주 ( Eun Joo Song ),김지민 ( Ji Min Kim ),서재석 ( Jae Seok Seo ),사금희 ( Keu 대한류마티스학회 2008 대한류마티스학회지 Vol.15 No.3
Objective: βig-h3 is an extracellular matrix protein, which is overexpressed in synovial tissues of rheumatoid arthritis (RA) similar to adhesive glycoproteins. We sought to evaluate the compensatory role of βig-h3 with adhesive glycoproteins in mediating the adhesion of fibroblast-like synoviocytes (FLS) and to confirm the inhibitory effect of YH18 peptide of the 2nd fas-1 domain in βig-h3-mediated adhesion. Methods: The adhesion of FLS isolated from synovial tissues of RA, was evaluated in 96 well microtiter plate coated with matrix proteins. Inhibitory effect of YH18 peptides from the 2nd and 4th fas-1 domains was estimated in βig-h3-mediated adhesion of FLS. Results: The adhesion of FLS on βig-h3 was weaker than that of fibronectin and vitronectin. The βig-h3-mediated adhesion was enhanced by the stimulation with phorbol myristate acetate (PMA), but not by cytokines and growth factors. Combination of fibronectin with βig-h3 synergistically enhanced the adhesion of FLS, in contrast to the additive effect of vitronectin combined with βig-h3. YH18 peptide of the 2nd fas-1 domain did not block the βig-h3-mediated adhesion of FLS. Conclusion: Our results reveal that βig-h3 may regulate the adhesion of FLS through the interaction with adhesive glycoproteins and confirm that the essential motifs mediating adhesion on βig-h3 are different according to the type of cells.
복수내 Fibronectin 치와 Adenosine Deaminase 치의 감별진단적 의의
권오종(Oh Jong Kwon),박승국(Soong Kook Park),박낭운(Rang Woon Park),김인산(In San Kim),조준승(Joon Seung Jo) 대한내과학회 1989 대한내과학회지 Vol.37 No.6
N/A The differential diagnosis of ascitic fluids remains to be solved. The aim of this study was to evaluate the diagnostic values of the fibronectin level and adenosine deaminase activity in differentiation of ascites. Ascitic fluid samples from 16 patients with malignancy, 5 patients with tuberculosis and 18 patients with liver cirrhosis were investigated. The mean ascitic fibronectin level was lower(p<0.01) in cirrhosis (32.8±16.7 μg/ml) than in malignancy (138.6±62.3 μg/ml) and tuberculosis (191.5±40.8 μg/ml) The mean value of ascitic adenosine deaminase activities was higher (P<0.01) in tuberculosis (68.1±37.2U/L) than in malignancy (11.4±5.9U/L) and cirrhosis (2.3±2.5 U/L) Fibronectin levels and adenosine deaminase activities did not show significant overlap among these 3 groups, while several other parameters analyzed simultaneously in these samples showed considerable overlap, By discriminant analysis, the fibronectin level was the best indicator for differentiation of tuberculous from cirrhotic ascities (% correctly classified 100) and the adenosine deaminase activity was the best indieator in the differential diagnosis of malignant and tuberculous ascities (% correctly classified=95.2). In differentiating the 3 groups, simultaneous determination of the fibronectin level and adenosine deaminase activity had the best discriminant ability (% correctly claseified=89.7) The results suggest that determination of both fibronectin level and adenosine deaminase activity in ascitic fluid may improve the accuracy of differential diagnosis of ascites.
βig-h3 절편-RGD 재조합단백의 만성염증성 관절염 치료효과
장지애 ( Ji Ae Jang ),강진희 ( Jin Hee Kang ),사금희 ( Keum Hee Sa ),한승우 ( Seung Woo Han ),서재석 ( Jae Seok Seo ),김경훈 ( Kyung Hoon Kim ),남언정 ( Eon Jeong Nam ),김인산 ( In San Kim ),강영모 ( Young Mo Kang ) 대한류마티스학회 2012 대한류마티스학회지 Vol.19 No.2
Objective. βig-h3 is a 68kDa extracellular matrix protein which is overexpressed in synovial tissues of rheumatoid arthritis (RA). Previous results proved that βig-h3 fragments are relevant to adhesion and migration of synovial fibroblast and angiogenesis through interaction with αvβ 3 integrin. We designed a recombinant βig-h3 protein consisting of a fas-1 domain and RGD motif and evaluated the therapeutic efficacy in RA. Methods. Inhibitory effect of adhesion and migration of NIH3T3 cell line was evaluated in 96 well microtiter and transwell plates coated with βig-h3. Clinical arthritis index was evaluated after treating CIA mice with MFK12. Immunohistochemical staining in synovial tissues were performed. Expression of transcripts and proteins of inflammatory mediators were analyzed by semi-quantitative RT-PCR and immunoblotting. Results. Recombinant protein consisted of 4th fas-1 domain truncated for H1 and H2 sequences and RGD peptide (MFK12), had M.W. of 10.4kDa. βig-h3 mediated adhesion and migration of NIH3T3 cell line were significantly inhibited in a dose-dependent manner. Arthritis severity and incidence were efficiently reduced when CIA mice were treated with MFK12 at 30 mg/kg/day compared with the control. Immunohistochemical staining of joint tissues in MFK12 treated mice exhibited reduced angiogenesis. In treated mice, expression of transcripts regarding inflammatory mediators was markedly suppressed and immunoblotting of ICAM-1 and RANKL from whole extract of hind paws also showed a significant reduction. Conclusion. This study shows that MFK12 is effective in treating RA, although further study is warranted to improve the therapeutic efficacy. Key Words. Rheumatoid arthritis, Inflammation, βig-h3, collagen-induced arthritis, Fas-1
Beta-lg H3을 이용한 사람 골수 중배엽 줄기세포의 연골분화 유도
이태규(Tae Kyu Lee),권굉우(Koing Woo Kwun),감상경(Sang Gyung Kim),김인산(In San Kim),조명래(Myung Rae Cho),조대원(Dae Won Cho) 대한정형외과학회 2004 대한정형외과학회지 Vol.39 No.4
목적: 사람 골수 중배엽 줄기세포(mesenchymal stem cells, MSC)의 연골분화 유도에서 TGF-beta를 대체할 수 있는 물질로서 Beta-Ig H3 (BigH3)의 가능성을 알아보고자 하였다. 대상 및 방법: 20명의 건강한 골수 공여자의 골수 흡인액에서 density gradient Ficoll-hypaque 분리방법으로 얻은 MSC를 알긴산을 지지체로 28일간 삼차원 배양하였다. 성장인자로 0.5 혹은 10ng/㎖의 transforming growth factor (TGF)-beta 1, TGF-beta 1+TGF-beta 3, BigH3을 첨가한 군에서 농도별, 시기별 세포 생존력, 총 콜라겐 함량, Glycosaminoglycan(GAG)함량을 측정하여 통계학적으로 비교하였다. 결과: TGF-beta 3은 배양 21일까지 유의하게 세포 생존력을 강화하였다(p=0.029). 10 ng/㎖ 농도일 때 TGF-beta 1+TGF-beta 3 첨가군과 BigH3 첨가군 간에는 세포생존력(p=0.197), 총 GAG 함량(p=0.253)에서 유의한 차이가 없었다. 총 콜라겐 함량은 l0ng/㎖ 농도의 경우 배양 21일까지 BigH3 첨가군이 TGF-beta 1+TGF-beta 3 첨가군보다 높았다(p=0.04l). 결론: 사람 골수의 MSC를 연골모세포로 성공적으로 분화 유도 하였으며, 이때의 배지에 첨가하는 성장인자로 TGF-beta 대신 국내에서 생산되는 BigH3을 대체하여 배양하여도 세포의 생존력과 단백당 함량 등 연골분화 유도의 외적 표지에 차이가 없음을 확인하였다. Purpose: This study was undertaken to compare the activity of beta-lg H3 (BigH3) and transforming growth factor (TGF)-beta on chondrogenesis by mesenchymal stem cells (MSCs). Materials and Methods: MSCs in human bone marrow aspirated from 20 healthy donors were isolated by density gradient Ficoll-hypaque separation and expanded in culture. MSC-alginate beads were prepared and incubated for 28 days in the presence of 0.5 or 10 ng/㎖ of TGF-beta 1, TGF-beta 1+TGF-beta 3 or BigH3. Cellular viability, total collagen and glycosaminoglycan (GAG) contents were measured and com¬pared. SPSS version 9.0 was used for the statistical analysis. Results: TGF-beta 3 significantly enhanced cell viability in beads by day 21 (p=0.029). No significant differences were found in terms of cell viability (p=0.197) or in total GAG content (p=0.253) between 10 ng/㎖ of TGF-beta 1+3 and 10 ng/㎖ of BigH3. Total collagen content was higher in the BigH3 added group on day 21 (p=0.041). Conclusion: The replacement of BigH3 instead of TGF-beta produced appropriate external signals indicating the chondrogenic differentiation of human bone marrow MSCs.