포도당, 인슐린 및 Angiotensin II가 흰쥐 대동맥평활근세포의 Plasminogen Activator Inhibitor-1 발현 및 성장에 미치는 영향

The Effects of Glucose, Insulin and Angiotensin II on Plasminogen Activator Inhibitor-1 Expression and Growth of Aortic Vascular Smooth Muscle Cell in Rats

  • 최세영 (계명대학교 의과대학 흉부외과) ;
  • 이인규 (계명대학교 의과대학 내과) ;
  • 한승세 (영남대학교 의과대학 흉부외과) ;
  • 김재현 (계명대학교 의과대학 흉부외과) ;
  • 박창권 (계명대학교 의과대학 흉부외과) ;
  • 이광숙 (계명대학교 의과대학 흉부외과) ;
  • 유영선 (계명대학교 의과대학 흉부외과) ;
  • 김기식 (계명대학교 의과대학 내과) ;
  • 김윤년 (계명대학교 의과대학 내과)
  • 발행 : 1999.04.01

초록

배경: PAI-1은 t-PA의 억제인자로서 섬유소융해계에 작용을 하여 혈전형성을 유발한다. PAI-1은 동맥경화된 혈관벽에서 분비가 된다. PAI-1의 증가는 동맥경화증의 위험인자가 되는 당뇨병과 고혈압이 동반된 환자에서 보이며 혈전증유발에 위험인자가 될 수 있다. 본 연구는 고혈당과 인슐린 및 angiotensin II가 PAI-1의 생성 및 평활근세포의 증식에 미치는 영향을 규명하고자 하였다. 대상 및 방법:흰쥐 대동맥평활근세포를 5.5 mM과 22 mM의 포도당 배양액을 사용하여 배양하였다. 배양액에 angiotensin II 및 인슐린을 농도 및 배양시간에 따라 첨가하여 Northern blotting방법으로 PAI-1 유전자발현을 나타내었다. 또한 세포 증식에 대한 포도당, 인슐린 및 angiotensin II의 영향을 규명하기 위하여 MTT assay를 사용하였다. 결과: 5.5 mM과 22 mM의 포도당 배양액에서 angiotensin II(100 nM)를 첨가하여 배양한 결과, 22 mM 포도당 배양액에서 PAI-1 mRNA 발현이 증가되었으며 angiotensin II 투여 4시간에 최고치에 도달하였고 6시간까지 지속되었다. 5.5 mM, 22 mM의 포도당 배양액에 angiotensin II의 농도를 0, 10, 100, 200 nM 투여하여 배양한 결과, PAI-1 mRNA의 발현은 angiotensin II 농도에 따른 증가를 보였으며 22 mM 포도당 배양액시 더욱 뚜렷하게 증가되었다. 배양액에 angiotensin II(100 nM)과 인슐린(100 nM)을 투여하여 배양한 결과, PAI-1 mRNA의 발현은 angiotensin II 단독으로 투여시 증가하였으나 인슐린을 첨가하였을 때는 감소하였다. 5.5 mM과 22 mM의 포도당 배양액에 1, 10, 100 nM의 인슐린과 1, 10, 100 nM의 angiotensin II를 첨가한 후 대동맥평활근세포의 성장속도를 비교한 결과, 5.5 mM보다 22 mM의 포도당이 든 배양액에서 대동맥평활근세포의 성장이 촉진되었으며, 인슐린 및 angiotensin II를 첨가한 경우도 대동맥평활근세포의 성장이 증가되었다. 결론:흰쥐 대동맥평활근세포에서 PAI-1 mRNA의 발현은 포도당 농도가 높을수록 증가되며 angiotensin II의 농도 및 배양시간에 따라 증가되고 인슐린 투여로 감소하였다. 또한 angiotensin II의 투여는 22 mM의 고농도 포도당 투여 후 증가된 PAI-1 mRNA 발현 증가를 더욱 증가시켜 PAI-1 mRNA 발현 증가에 상승작용이 있음을 알 수 있다. 그리고 22 mM의 고농도 포도당, 인슐린 및 angiotensin II는 흰쥐의 대동맥평활근세포의 성장을 촉진시켰다.

Background: Plasminogen activator inhibitor-1(PAI-1) is known as the primary physiological inhibitor of tissue-type plasminogen activator(t-PA) in the plasma, and is present within the atherosclerotic vessels. Increased plasma levels of PAI-1 are one of the major disturbances of the hemostatic system in patients with diabetes and/or hypertension, and may have multiple interrelations with the important risk factors in the development of atherosclerosis. This study was performed to determine whether altered gene expression of PAI-1 occurs within the arterial wall, and thereby potentially contributing to the increase of cardiovascular risks associated with diabetes and/or hypertension. Material and Method: The aortic vascular smooth muscle cells of the rat were exposed to 22 mM glucose, angiotensin II, and insulin increased PAI-1 mRNA expression with the use of Northern blotting were examined. Also examined were the effects of 22 mM glucose, angiotensin II and insulin on the growth of the rat's aortic smooth muscle cells by using MTT assay. Result: Twenty-two mM glucose treatment increased the PAI-1 mRNA expression in a time- and dose-dependent manner. Aniotensin II treatment synergistically increased the glucose-induced PAI-1 mRNA expression. In contrast, addition of insulin attenuated the increase of 22 mM glucose and angiotensin II induced PAI-1 mRNA expression. Furthermore, treatment of 22 mM glucose, angiotensin II and insulin resulted in a significant increase in cell numbers. This study demonstrated that 22 mM glucose and angiotensin II have a synergistic effect in stimulating the PAI-1 mRNA expression and in the cell growth of the rat's aortic smooth muscle cells. Conclusion: Elevation of glucose and angiotensin II may be important risk factors in impairing fibrinolysis and developing atherosclerosis in diabetic patients.

키워드

참고문헌

  1. Hypertension v.19 no.6 Induction of endothelin-I gene by angiotensin and vasopressin in endothelial cells Imai T;Hirata Y;Emori T;Yanagisawa M;Masaki T;Marumo F
  2. Hypertension v.19 no.SUP. 2 Stimulation of endothelial cell prostaglandin production by angiotensin peptides. Characterization of receptors Jaiswal N;Diz DI;Chappell MC;Khosla MC;Ferrario CM
  3. Hypertension v.13 no.4 Angiotensin Ⅱ-stimulated protein synthesis in cultured vascular smooth muscle cell Berk BC;Vekshtein V;Gordon HM;Tsuda T
  4. Hypertension v.19 no.SUP. 2 Possible role of the vascular renin-angiotensin system in hypertension and vascular hypertrophy Morishita R;Higaki J;Miyazaki M;Ogihara T
  5. J Clin Invest v.91 no.5 Multiple autocrine growth factors modulate vascular smooth mucle cell growth response to angiotensin Ⅱ Itoh H;Mukoyma M;Pratt RE;Gibbons GH;Dzau VJ
  6. Cell v.71 no.1 Molecular basis of human hypertension: role of angiotensinogen Jeunemaitre X;Soubrier F;Kotelevtsew YV;Lifton RP;Williams CS;Charru A;Hunt SC(et al)
  7. N Engl J Med v.330 no.23 Linkage of the angiotensinogen gene to essential hypertension Caulfield M;Lavender P;Farrall M;Munroe P;Lawson M;Turner P;Clark AJL
  8. N Eng J Med v.330 no.23 Association between a deletion polymorphism of the angiotensin-converting-enzyme gene and left ventricular hypertorphy Schunkert H;Hense HS;Holmer SR;Stender M;Perz S;Keil U;Lorell BH(et al)
  9. Lancet v.342 no.8879 Angiotensin-converting enzyme DD genotype in patients with ischaemic or idiopathic dilated cardiomyopathy Raynolds MV;Brestow MR;Bush EW;Abraham WT;Lowes BD;Zisman LS;Taft CS(et al)
  10. Nature v.359 no.6393 Deletion polymorphism in the gene for angiotensin-converting enzyme is a potent risk factor for myocardial infarction Cambien F;Poirier O;Lecert L;Evans A;Cambou JP;Arveiler D;Luc G(et al)
  11. Diabetes v.43 no.5 Genetic predis-position to diabetic nephropathy. Evidence for a role of the angiotensin I-converting enzyme gene Doria A;Warram JH;Krolewski AS
  12. Diabetes v.43 no.3 Relationships between angiotensin I converting enzyme gene polymorphiam, plasma levels and diabetic retinal and cells renal complications Marre M;Bernadet P;Gallois Y;Savagner F;Guyene TT;Hallab M;Cambien F(et al)
  13. Diabetes v.41 no.8 Increased expression of tissue plasminogen activator and its inhibitor and reduced fibrinolytic potential of human endothelial cells cultured in elevated glucose Maiello M;Boeri D;Podesta F;Cagliero E;Vichi M;Odetti P;Adezati L(et al)
  14. J Biol Chem v.265 no.4 Isolation and characterization of the rat plasminogen activator inhibitor-I gene Bruzdzinski CJ;Riordan-Johnson M;Nordby BC;Suter SM;Gelehrter TD
  15. Circ Res v.78 no.3 Migration of arterial wall cells. Expression of plasminogen activators and inhibitors in injured rat arteries Reidy MA;Irvin C;Lindner V
  16. J Clin Res v.88 no.4 The plasminogen activator/plasmin system Vassalli JD;Sappino AP;Belin D
  17. Semin Thormb Hemost v.18 no.1 Plasminogen activator inhibitor type I: biochemistry and evidence for modulation of fibrinolysis in vivo Krishnamurti C;Alving BM
  18. Thromb Haemost v.57 no.1 Deficient t-PA release and elevated PA inhibitor levels in patients with spontaneous or recurrent deep vein thrombosis Juhan-Vague I;Valadier J;Alessi MC;Aillaud MF;Ansaldi J;Pillip-Joet C;Holvoet P(et al)
  19. Br J Med v.291 no.6495 Plasminogen activator inhibitor in the blood of patients with coronary artery disease Paramo JA;Colucci M;Collen D;van de Werf F
  20. N Eng J Med v.313 no.25 Increased plasma levels of a rapid inhibitor of tissue plasminogen activator in young survivors of myocardial infartion Hamsten A;Wiman B;De Faire U;Blomback M
  21. Circulation v.96 no.2 Effects of ramipril on plasma fibrinolytic balance in patients with acute anterior myocardial infarction. HEART study investigators Vaughan DE;Rouleau JL;Ridker PM;Arnold JM;Menapace FJ;Pfeffer MA
  22. Nature(Lond) v.346 no.1 Development of venous occlusions in mice transgenic for the plasminogen activator inhibitor-1 gene Erickson LA;Fici GA;Lund JE;Boyle TP;Polites HG;Marotti KR
  23. J Clin Invest v.92 no.6 Plasminogen activator inhibitor-1 gene-deficient mice Ⅱ. Effects on Hemostasis, thrombosis, and thrombolysis Carmeliet P;Stassen JM;Schoonjans L(et al)
  24. Thromb Haemost v.61 no.3 Increased plasminogen activator inhibitor activity in non insulin dependent diabetic patients: relationship with plasma insulin Juhan-Vague I;Roul C;Alessi MC;Ardissone JP;Heim M;Vague P
  25. J Clin Invest v.83 no.4 Induction of platelet-derived growth factor A-chain and c-myc gene expressions by angiotensin Ⅱ in cultured rat vascular smooth muscle cells Naftilan AJ;Pratt RE;Dzau VJ
  26. J Biol Chem v.164 no.1 Angiotensin Ⅱ induces c-fos mRNA in aortic smooth muscle Taubman MB;Berk BC;Izumo S;Tsuda T;Alexander RW;Nadal-Ginard B
  27. Am J Hypertens v.6 no.6 Synergic action of angiotensin Ⅱ, insulin-like growth factor-I, and transforming growth factor-beta on platelet-derived growth factor-BB, basic fibroblast growth factor, and epidermal growth factor-induced DNA synthesis in vascular smooth muscle cells Ko Y;Stiebler H;Nickenig G;Wieczorek AJ;Vetter H;Sachinidis A
  28. J Biol Chem v.268 no.22 Angiotensin Ⅱ regulated insulinlike growth factor I gene expression in vascular smooth muscle cells Delafontaine P;Low H
  29. Am J Hypertens v.3 no.11 New angiotensin converting enzyme inhibitors. Their role in the management of hypertension Laragh JH
  30. Science v.245 no.4919 Inhibitors of angiotensin converting enzyme prevent myointimal proli-feration after vascular injury Powell JS;Clozel JP;Muller RK(et al)