사람의 제2형 당뇨병성 신증의 신조직내 TGF-β-inducible Gene-h3의 발현과 유용성-면역조직화학적 분석- = Renal Expression and Usefulness of TGF-β-inducible Gene-h3 in Human Type II Diabetic Nephropathy -Immunohistochemical Analysis-

국문 초록 (Abstract)

목 적:신장 베타 변형성장인자 (TGF-β)계의 활성화가 당뇨병성 신증의 주요 인자로 알려져 있다. TGF-β는 비활성형으로 생성되어 생물학적 활성도를 갖는 형태로 변형되는데, 최근에 TGF-β inducible gene-h3 (βig-h3)가 TGF-β에 의해 유도되는 유전자로써 알려지면서 TGF-β의 활성도를 시사하는 유전자로 인정받고 있다.
방 법:금번 실험에서는 제2형 당뇨병성 신증의 전형적인 조직소견과 임상소견을 보이는 사람을 대상으로 신 조직 내 βig-h3 발현을 보았고, TGF-β1과 그 제2형 수용체 및 Smad3에 대해서도 면역조직학적 염색을 시행하여 비교 분석하였다.
결 과:이번 분석에서 βig-h3가 신조직내에 분포하는 양상을 보면, 당뇨병성 신증의 11예 중 5예는 사구체에, 7예는 세뇨관에 염색이 증가되어 있는 소견이 있었고, 정상 대조군의 경우 사구체나 세뇨관 모두 전혀 발현되지 않았다. TGF-β1의 염색 결과, 정상 조직에서는 전혀 염색되지 않는 양상을 보인 반면, 당뇨병성 신증에서는 사구체에서는 11예 중 3예와 세뇨관에서는 11예 중 1예에서 과염색된 것을 관찰할 수 있었으나, 통계학적인 의의는 없었다. TβRII의 염색 결과, 세뇨관 발현은 정상과 별 차이가 없었으나, 사구체는 당뇨병성 신증인 경우 11예 중 6예에서 과염색된 반면, 정상에서는 3예 중 1예에서 과염색되어 당뇨병성 신증에서 증가되어 있었다. Smad3의 염색 결과, 당뇨병성 신증과 정상 모두 사구체 및 세뇨관에 발현되었고, 단위 사구체 당 염색된 세포의 수로 비교하여 당뇨병성 신증에서 49.1±0.3으로 정상 (40.9±0.8)에 비해 약간 증가하였다.

다국어 초록 (Multilingual Abstract)

Background:Activation of transforming growth factor-β (TGF-β) system has been implicated in the pathological change of diabetic nephropathy such as renal hypertrophy and accumulation of extracellular matrix. In tissues, TGF-β is secreted as a biologically inactive complex requiring in vivo activation. Thus, increased TGF-β1 mRNA or protein may not necessarily reflect parallel changes in TGF-β1 biologic activity. TGF-β1 inducible gene-h3 (βig-h3) is a novel gene uniquely up-regulated by active TGF-β1.
Methods:For evaluating the βig-h3 protein expression in human diabetic nephropathy, we examed the expression of βig-h3 protein, TGF-β1, TGF-β type II receptor (TRII) and Smad protein intranuclear translocation by immunohistochemistry in human diabetic nephropathy tissues (n=11) and normal renal tissue (n=3).
Results:The βig-h3 protein was expressed in diabetic tubular epithelium (diabetes vs. normal 7/11 vs. 0/3) and diabetic glomerulus (diabetes vs. normal 5/11 vs. 0/3). The tubular expression was stronger than the interstitial expression. The TGF-β1 was expressed in diabetic tubular epithelium (diabetes vs. normal 1/11 vs. 0/3) and diabetic glomerulus (diabetes vs. normal 3/11 vs. 0/3). The TGF-β type II was expressed more in diabetic glomerulus (diabetes vs normal 6/11 vs. 1/3). But in the tubule, the expression didn't show any significant difference. The number of intranuclear translocation of Smad protein in glomerulus (diabetes vs normal 49.1±0.3 vs. 40.1±0.8) was increased in diabetes, but the tubular manifestation was not significant.
Conclusion:We propose that TGF-β system mediates human diabetic nephropathy through βig- h3 protein expression. The βig-h3 protein expression could be a useful marker expecting of disease activity and progress.

참고문헌 (Reference)

• 1 "당뇨병성 신증 : 당뇨병학" 고려의학 1998
• 2 "cDNA cloning and sequence analysis of beta ig-h3,a novel gene induced in a human adenocarcinoma cell line after treatment with transforming growth factor- beta" 11 : 511-522, 1992
• 3 "a transforming growth factor- beta-responsive gene encoding a secreted protein that inhibits cell attachment in vitro and suppresses the growth of CHO cells in nude mice" 571-584, 1994
• 4 "Transforming growth factor- beta1 and tumor growth factor-beta-inducible gene-H3 in nonrenal transplant cyclosporine nephropathy" 72 : 1826-1829, 2001
• 5 "The key role of the transforming growth factor-beta system in the pathogenesis of diabetic nephropathy" 23 : 471-481, 2001
• 6 "The effects of high glucose concentration, angiotensin II and its blockers on the production of IL-6 and fibronectin in cultured human mesangial cells" 61 : 399-408, 2001
• 7 "The effects of high glucose concentration on angiotensin II- or transforming growth factor-beta-induced DNA syn thesis,hypertrophy and collagen synthesis in cultured rat mesangial cells" 37 : 302-311, 1996
• 8 "Stimulation of collagen gene expression and protein synthesis in murine mesangial cells by high glucose is mediated by autocrine activation of transforming growth factor-beta" 93 : 536-542, 1994
• 9 "Smad-dependent and Smad-independent pathways in TGF-β family signalling" 425 : 577-584, 2003
• 10 "Serum levels of transforming growth factor(TGF)-beta1 in type 2 diabetic patients" 22 : 522-530, 1998
• 11 "Renal expression of transforming growth factor-beta inducible gene- h3(beta ig-h3)in normal and diabetic rats" 54 : 1052-1062, 1998
• 12 "Long-term prevention of renal insufficiency, excess matrix gene expression, and glomerular mesangial matrix expansion by treatment with monoclonal antitransforming growth factor-beta antibody in db/db diabetic mice" 97 : 8015-8020, 2000
• 13 "Increased glomerular and tubular expression of transforming growth factor-b1, its type II receptor, and activation of the smad signaling pathway in the db/db mouse" 158 : 1653-1663, 2001
• 14 "Glycated albumin stimulates TGF-β1 production and protein kinase C activity in glomerular endothelial cells" 59 : 673-681, 2001
• 15 "Expression of TGF-β-βig-h3 is up- regulated in the diabetic rat kidney and human proximal tubular epithelial cells treated with high glucose" 64 : 1012-1021, 2003
• 16 "Effects of graded control of blood glucose with insulin on the progression of experimental diabetic nephropathy" 18 : 894-903, 1999
• 17 "Effect of transforming growth factor-induced gene product, beta ig-h3 on proliferation, migration, and adhesion of aortic smooth muscle cells cultured in high glucose" 26 : 286-295, 2002
• 18 "Effect of transforming growth factor-B1 and platelet derived growth factor on synthesis and gene expression of collagen and non-collagen protein in aortic smooth muscle cells cultured in different concentrations of insulin and glucose" 23 : 518-529, 1999
• 19 "Effect and mechanism of high glucose level on the expression of an adhesion protein, beta ig-h3, and cellular function in endothelial cells" 27 : 323-331, 2003
• 20 "Differential display of human marrow stromal cells reveals unique mRNA expression patterns in response to dexamethasone" 76 : 231-243, 1999
• 21 "Beta-ig.Molecular cloning and in situ hybridization in corneal tissues" 38 : 893-900, 1997
• 22 "Beta IG-H3, a novel secretory protein inducible by transforming growth factor-beta, is present in normal skin and promotes the adhesion and spreading of dermal fibroblasts in vitro" 104 : 844-849, 1995
• 23 "Beta (ig) H-3 : why should we know about this molecule?" 72 : 1725-1726, 2001
• 24 "A TGF- beta-inducible cell adhesion molecule,betaig-h3,is downregulated in melorheostosis and involved in osteogenesis" 77 : 169-178, 2000