간경변 백서 모델에서 아데노바이러스 벡터를 이용한 Matrix Metalloproteinase-3 유전자 치료
Gene therapy via adenovirus vector carrying matrix metalloproteinase-3 (MMP-3) in a rat model of liver cirrhosis
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
목적 : 간경변은 활성화된 간성상 세포에 의해 이상 증식된 세포외기질이 축적되는 것을 특징으로 하며 matrix metalloproteinase (MMP)에 의해 분해되는 것으로 보고되고 있다. 본 연구의 목적은 간경변 모델 흰쥐에서 MMP-3를 발현하는 아데노바이러스를 이용한 유전자 치료로 간섬유화가 경감되는지를 알아보고자 하였다. 방법 : 몸무게 200∼250 g의 6주령된 수컷 백서에 8주 동안 DMN을 복강에 주사하여 간경변 동물 모델을 만들었다. 일주일 후에 AdMMP3.GFP, Ad.GFP, PBS를 문맥에 주사한 후 매주마다 백서를 희생시켜 조직을 채취하였다. 간경변의 개선정도는 활성형의 간성상 세포 지표인 a-SMA, Desmin, TGF RⅡ의 면역화학적 염색, 간세포 성장 표지자인 PCNA 및 dry liver weight 측정을 통해 분석하였다.
Abstract
Background : Liver cirrhosis is characterized by fibrous scarring and hepatocellular regeneration. Matrix metalloproteinases (MMPs) comprise a family of zinc-dependent enzymes that degrade the extracellular matrix (ECM) components. This study examined whether or not gene delivery of human MMP-3 can attenuate established liver cirrhosis in a rat. Methods : Rat liver cirrhosis was induced by an intraperitoneal injection of dimethylnitrosamine (DMN) three times a week for 8 weeks. The rats were infected once with either a recombinant adenovirus, AdMMP3.GFP, or a control adenovirus, Ad.GFP, into a portal vein and followed up for 3 weeks. In the rat liver tissues, the collagen content, histopathology and immunohistochemical staining were measured. Results : Liver fibrosis in the DMN induced cirrhotic rat was attenuated along with a diminished hydroxyproline content and increased dried liver weight after the gene delivery of AdMMP3.GFP. In addition, the number of activated hepatic stellate cells was lower whereas the proliferation of hepatocytes, which was confirmed by immunohistochemical staining using anti-proliferating cell nuclear antigen (PCNA) antibody, was observed in the AdMMP3.GFP infected rats, suggesting that human MMP-3 stimulated hepatocyte proliferation. Conclusions : These results suggest that the gene transfer of human MMP-3 in the liver attenuates established fibrosis and induces hepatocyte proliferation. Therefore, gene therapy using MMP-3 in liver cirrhosis might be a promising therapeutic option in the future.(Korean J Med 71:609-619, 2006)
참고문헌 (22)
- "Molecular regulation of hepatic fibrosis an integrated cellular response to tissue injury", 275, pp.2247~2250 (2000)
- "Activation of Ito cells involves regulation of AP-1 binding proteins and induction of type 1 collagen gene expression", 304, pp.817~824 (1994)
- "Kupffer cells from carbon tetrachloride- injured rat livers produce chemotactic factors for fibroblast and monocytes:the role of tumor necrosis factor alpha", 14, pp.895~900 (1991)
- "Expression of matrix metalloproteinases and their inhibitors during hepatic tissue repair in the rat", 113, pp.443~453 (2000)
- "Serum matrix metalloproteinase-3 (stromelysin-1) concentration in patients with chronic liver disease.", 31, pp.474~481 (1999)
- "Delivery of matrix metalloproteinase-1 attenuates established liver fibrosis in the rat", 124, pp.445~458 (2003)
- "Treatment with human metalloproteinase-8 gene delivery ameliorates experimental rat liver cirrhosis", 126, pp.1122~1133 (2004)
- "Significance of coexpression of urokinase-type plasminogen activator, and matrix metalloproteinase 3 (stromelysin) and 9 (gelatinase B) in colorectal carcinoma.", 93, pp.211~218 (2000)
- "Messenger RNA for myosin polypeptides:isolation from single myogenic cell cultures", 10, pp.265~273 (1977)
- "Isolation of RNA using guanidinium salts", 152, pp.219~227 (1987)
- "백서 눈에서 아데노바이러스 벡터를 매개로 한 Stromelysin 유전자의 발현", 42, pp.378~385 (2001)
- "Structure-related cytotoxicity and anti-hepatofibric effect of asiatic acid derivatives in rat hepatic stellate cell-line, HSC-T6", 27, pp.512~517 (2004)
- "The sympathetic nervous system promotes carbon tetrachloride-induced liver cirrhosis in rats by suppressing apoptosis and enhancing the growth kinetics of regenerating hepatocytes", 36, pp.111~120 (2001)
- "A prospective randomized trial of colchicine in prevention of liver cirrhosis in chronic hepatitis B patients", 10, pp.961~966 (1996)
- "Activation mechanisms of matrix metalloproteinase", 378, pp.151~160 (1997)
- "Matrix metalloproteinase and diseases of the CNS", 21, pp.75~80 (1998)
- "Serum tissue inhibitor of metalloproteinase in patients with chronic liver disease and with hepatocellular carcinoma", 218, pp.47~58 (1993)
- "Preventive and therapeutic effects in rats of hepatocyte growth factor infusion on liver fibrosis", cirrhosis, 1997, pp.26, 81~89
- "Selective inhibition of hepatic collagen accumulation in experimental liver fibrosis in rats by a new prolyl 4-hydroxylase inhibitor", 28, pp.404~411 (1998)
- "시험관내 공동배양법을 이용한 C형 간염 바이러스 Core 단백이 간섬유화 기전에 미치는 영향", 42, pp.400~408 (2003)
- "Expression of insulin-like growth factor I by activated hepatic stellate cells reduces fibrogenesis and enhances regeneration after liver injury", 54, pp.134~141 (2005)
- "Insulin-like growth factor 1 blocks collagen release and down regulates matrix metalloproteinase-1 and -13 mRNA expression in bovine nasal cartilage stimulated with oncostatin M in combination with interleukin 1alpha", 60, pp.254~261 (2001)