2000 년도산학협동연구비 Korean Circulation J 2002;32 12 : 생쥐의 Adriamycin 유발심근증모델에서 IGF-1 의 심근보호효과 박철수 윤호중 조은주 오용석 백상홍 이정화강진형 정욱성 채장성 김재형 최규보 홍순조 Cardi

Size: px

Start display at page:

Download "2000 년도산학협동연구비 Korean Circulation J 2002;32 12 : 생쥐의 Adriamycin 유발심근증모델에서 IGF-1 의 심근보호효과 박철수 윤호중 조은주 오용석 백상홍 이정화강진형 정욱성 채장성 김재형 최규보 홍순조 Cardi"

병길 석
5 years ago
Views:

1 2000 년도산학협동연구비 Korean Circulation J 2002;3212: 생쥐의 Adriamycin 유발심근증모델에서 IGF-1 의 심근보호효과 박철수 윤호중 조은주 오용석 백상홍 이정화강진형 정욱성 채장성 김재형 최규보 홍순조 Cardioprotective Effect of IGF-1 in Mouse with Adriamycin Induced Cardiomyopathy Chul-Soo Park, MD, Ho-Joong Youn, MD, Eun-Joo Cho, MD, Yong-Seok Oh, MD, Sang-Hong Baek, MD, Jung-Hwa Lee, MD, Jin-Hyoung Kang, MD, Wook-Sung Chung, MD, Jang-Seong Chae, MD, Jae-Hyung Kim, MD, Kyu-Bo Choi, MD and Soon-Jo Hong, MD Department of Internal Medicine, Department of Biochemistry, College of Medicine, The Catholic University of Korea, Seoul, Korea ABSTRACT Background and ObjectivesThe purpose of this study was to investigate the apoptotic pathway, and the effect and mechanism of IGF-1 insulin like growth factor 1 as a therapeutic agent, in Adriamycin ADR induced cardiomyopathy. Materials and MethodsWe divided 17 mice into ADR n6, ADR IGF-1 n6 and saline n5 groups. The following were injected into the intraperitoneal cavity for the specified periods 2.5 mg/kg of ADR every week for 6 weeks, 0.1 mg/kg of IGF-1 in the 5th and 6th weeks and 10 ml/kg of saline every week for 6 weeks. Transthoracic echocardiography, with a 15MHz linear array, was performed before and after injection of the drugs. A TUNEL assay and immunohistochemical staining for Bax, bcl-2, capase-8 and 9, and western blotting for Bax and bcl-2 were performed on the myocardium. ResultsThe fractional shortening decreased % before vs % after, p0.05 and the left ventricular end systolic dimension increased mm before vs mm after, p0.05 following the injection of the ADR, but there were no interval changes in the mice treated with saline or ADRIGF-1. The TUNEL assay showed a higher apoptotic index in the mice treated with ADR than in those treated with saline or ADRIGF % in ADR, % in control, 225% in ADRIGF-1, p0.05. The immunohistochemical staining and western blot showed an increase in the expression of Bax, and a decrease in the expression of bcl-2 in the mice treated with ADR than in those treated with ADRIGF-1 or saline. ConclusionThe apoptosis caused by the increased expression of Bax, and the decreased expression of bcl-2, was an important pathogenetic mechanism, and the IGF-1 prevents the progression of cardiomyopathy by attenuating the expressions of Bax and bcl-2 in ADR induced cardiomyopaththy. Korean Circulation J 2002;3212: KEY WORDSDoxorubicinCardiomyopathy, congestiveapoptosisinsulin-like growth factor I. 1116

2 서론 대상동물 재료및방법 - 심초음파를이용한심근증의평가 Fig. 1. The schedule of echocardiographic monitoring and injection of saline, IGF-1, ADR. IGF-1insulin like growth factor, ADRadriamycin. 1117

3 심근조직의획득및처리 Hematoxylin-eosin 염색및 TUNEL assay 면역조직학적염색 Western blotting 통계적검증 - 결과 심초음파를통한심근증의평가 Korean Circulation J 2002;3212:

4 세포자멸의확인 - Fig. 2. M mode echocardiographic images of left ventricle in mice treated with saline A, ADR alone B, IGF-1 and ADR C. Upper panels are baseline echocardiographic images and lower panels are those after injecting saline or drugs. Panel B showed significantly decreased wall motion and fractional shortening after injection of Adriamycin but in Panel A and C, there were no significant changes after injection drugs. LVleft ventricle, ADRadriamycin. Fig. 3. Hemodynamic and morphologic changes in mice treated with saline A, ADR alone B, and IGF-1 and ADR C. Panel B showed significantly increased LVIDs and decreased FS after injection of ADR. IGF-1insulinlike growth factor 1, ADRadriamycin, FSfractional shortening, LVIDd/LVIDsleft ventricular interventricular dimension at diastole/systole. p

Apoptotic nuclei are stained in brown (white arrow). Panel B showed higher rate of positive cells than Panel (A) and (C).

Bax and bcl-2 immunohistochemical staining of myocardium from mice treated with saline (A), Adriamycin alone (B), IGF-1 and Adriamycin (C).

Panel B showed higher rate of positive cells in Bax staining and lower rate of positive cells in bcl-2 staining than panel C.

5 A B C Fig. 4. Histological sections of myocardium from mice treated with saline (A), Adriamycin alone (B) and IGF-1 and Adriamycin (C) stained with TUNEL assay. Apoptotic nuclei are stained in brown (white arrow). Panel B showed higher rate of positive cells than Panel (A) and (C). IGF-1 insulin-like growth factor I, TUNEL terminal deoxynucleotidyl transferase-mediated end labelling (magnification, 400). Fig. 5. Bax and bcl-2 immunohistochemical staining of myocardium from mice treated with saline (A), Adriamycin alone (B), IGF-1 and Adriamycin (C). Positive staining is shown by red color in cytoplasm. Panel B showed higher rate of positive cells in Bax staining and lower rate of positive cells in bcl-2 staining than panel C. IGF-1 insulin-like growth factor I (magnification, 400). 인해 정확한 세포자멸을 확인하기 위해 TUNEL assay 조직학적 염색 를 시행하여 자멸사 세포지수를 비교하였다. 세 군의 자 Bax의 활성도는 Adriamycin 투여군에서 Adriamy- 멸사 세포지수는 대조군 0.1±0.03%, Adriamycin 투 cin, IGF-1 병합 투여군이나 대조군에 비해 현저히 증 여군 46±8%, Adriamycin IGF-1 병합투여군 22± 가되어 있었으며 bcl-2의 활성도는 IGF-1, Adriamy- 5%로 Adriamycin군에서 다른 군에 비해 유의한 세포 cin 병합 투여군에서 대조군이나 Adriamycin 투여군에 자멸의 증가를 확인할 수 있었다(p<0.05)(Fig. 4). 비해 현저히 증가되어 있었다. caspase-8과 caspase Korean Circulation J 2002; 32(12):

Panel B showed denser band of Bax and looser band

6 Fig. 6. The results of western blot of Actin, bcl-2, and Bax from mice myocardium treated with saline A, Adriamycin alone B, IGF-1 and Adriamycin C. Panel B showed denser band of Bax and looser band of bcl-2 than Panel A and C. IGF-1insulin-like growth factor I. 각군의 Western blotting결과 - 고 찰

7 요약 배경및목적 : 방법 : 결과 : - Korean Circulation J 2002;3212:

8 - - - 결론 : 중심단어 REFERENCES 1) Rosen GM, Halpern HJ. Spin trapping biologically generated free radicals correlating formation with cellular injury. Methods Enzymol ) Singal PK, Iliskovic N, Li T, Kumar D. Adriamycin cardiomyopathy phthophysiology and prevention. FASEB J ) Wang L, Ma W, Markovic R, Chen JW, Wang PH. Regulation of cardiomyocyte apoptotic signaling by insulin-like growth factor I. Circ Res ) Kumar D, Kirshenbaum LA, Li T, Danelisen I, Singal PK. Apoptosis in adriamycin cardiomyopathy and its modulation by probucol. Antioxid Redox Signal ) Arola OJ, Saraste A, Pulkki K, Kallajoki M, Parvinen A, Voipio-Pulkki LM. Acute doxorubicin cardiotoxicity involves cardiomyocyte apoptosis. Cancer Res ) Sawyer DB, Fukazawa R, Arstall MA, Kelly RA. Daunorubicin-induced apoptosis in rat cardiac myocytes is inhibited by dexrazoxane. Circ Res ) Lee WL, Chen JW, Ting CT, Ishiwata T, Lin SJ, Korc M, Wang PH. Insulin-like growth factor 1 improves cardiovascular function and suppresses apoptosis of cardiomyocytes in dilated cardiomyopathy. Endocrinology ) Yamamura T, Otani H, Nakao Y, Hattori R, Osako M, Imamura H. IGF-1 differentially regulates Bcl-xL and Bax and confers myocardial protection in the rat heart. Am J Physiol Heart Circ Physiol H ) Singal PK, Deally CM, Weinberg LE. Subcellular effects of adriamycin in the heart a consice review. J Mol Cell Cardiol ) Sinha BK, Katki AG, Batist G, Cowan KH, Myers CE. Adriamycin-stimulated hydroxyl radical formation in human breast tumor cells. Biochem Pharmacol ) Muller I, Jenner A, Bruchelt G, Niethammer D, Halliwell B. Effect of concentration on the cytotoxic mechanism of doxorubicin apoptosis and oxidative DNA damage. Biochem Biophys Res Commun ) Yen HC, Oberley TD, Gairola CG, Szweda LI, St Clair DK. Manganese superoxide dismutase protects mitochondrial complex I against adriamycin induced cardiomyopathy in transgenic mice. Arch Biochem Biophys ) Sawyer DB, Fukazawa R, Arstall MA, Kelly RA. Daunorubicin-induced apoptosis in rat cardiac myocyte is inhibited by dexrazoxane. Circ Res ) Nakamura T, Ueda Y, Juan Y, Katsuda S, Takahashi H, Koh E. Fas-mediated apoptosis in adriamycin-induced cardiomyopathy in rats. Circulation ) Ambler GR, Johnson BM, Maxwell L, Gavin JB, Gluckman PD. Improvement of doxorubicin induced cardiomyopathy in rats treated with insulin-like growth factor I. Cardiovasc Res ) Pecherskaya A, Solem M. IGF-1 activates PKC alphadependent protein synthesis in adult rat cardiomyocytes. Mol Cell Biol Res Commun ) Wu W, Lee WL, Wu YY, Chen D, Liu TJ, Jang A, Sharma PM, Wang PH. Expression of constitutively active phosphati-dylinositol 3-kinase inhibits activation of caspase- 3 and apoptosis of cardiac muscle cells. J Biol Chem ) Hong F, Kwon SJ, Jhun BS, Kim SS, Ha J, Kim SJ, Sohn NW, Kang C, Kang I. Insulin-like growth factor-1 protects H9c2 cardiac myoblasts from oxidative stress-induced apoptosis via phosphatidylinositol 3-kinase and extracellular signal-regulated kinase pathways. Life Sci

( )Kju269.hwp

( )Kju269.hwp 만성세균성전립선염모델흰쥐에서 의항염효과 Anti-inflammatory Effect of Lycopene on Chronic Bacterial Prostatitis Rat Model Cho Hwan Yang, Dong Wan Sohn, Yong-Hyun Cho From the Department of Urology, The Catholic University