title: Countervailing effects of rapamycin (sirolimus) on nuclear factor-kappa B activities in neointimal and medial smooth muscle cells.
abstract: OBJECTIVE
  Local application of rapamycin (sirolimus) by drug-eluting stents prevents lumen obliteration after angioplasty by inhibition of neointimal hyperplasia. The effects of rapamycin on neointimal smooth muscle cells (niSMC) which are responsible for the occurrence of restenosis have not been investigated so far.
  
  
  RESULTS
  Rat niSMC and medial SMC (mSMC) were obtained from balloon catheter-injured arteries. The niSMC exhibited higher basal NF-kappaB activity and TNF-alpha mRNA levels. Nuclear protein binding to NF-kappaB-DNA was attenuated in niSMC by incubation with rapamycin (0.1 and 1 microg/ml) for 24 and 48 h. In contrast in mSMC, 0.1 microg/ml rapamycin had no effect and at 1 microg/ml even increased nuclear protein binding to NF-kappaB-DNA. After 12 h incubation, rapamycin (0.001-10 microg/ml) induced IkappaB-alpha protein in niSMC, whereas in mSMC it stimulated IkappaB-alpha at much lower levels. Prolonged rapamycin treatment (1 microg/ml for 72 h) had no effect on TNF-alpha mRNA level and NF-kappaB activity in niSMC, whereas it led to their increase in mSMC. Vascular endothelial growth factor (VEGF) secretion was higher in mSMC than in niSMC; rapamycin decreased VEGF levels in both cell types. Ultrastructural analysis suggested that rapamycin caused early signs of degeneration in niSMC, but enhanced protein synthesis in mSMC.
  
  
  CONCLUSIONS
  This study shows that rapamycin influences the inflammatory phenotypes of SMC in opposite directions: it reduces the high basal NF-kappaB activity in niSMC and enhances NF-kappaB activity and TNF-alpha expression in mSMC. In addition, rapamycin inhibits VEGF production regardless of the phenotype of SMC. These findings shed light on molecular mechanisms and structural changes underlying therapeutic applications of rapamycin in prevention of restenosis, inhibition of chronic transplant arteriosclerosis and reduction of secondary malignoma formation due to immunosuppression.
authors.affiliation: Clinical Department of Cardiology, Medical University Innsbruck, Anichstrasse 35, A-6020 Innsbruck, Austria. wolfgang.dichtl@uibk.ac.at
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Dichtl
authors.forename: Eva-Maria
authors.initials: EM
authors.lastname: Stocker
authors.forename: Klaudia
authors.initials: K
authors.lastname: Mistlberger
authors.forename: Paul
authors.initials: P
authors.lastname: Debbage
authors.forename: Zhong-qun
authors.initials: ZQ
authors.lastname: Yan
authors.forename: Hannes F
authors.initials: HF
authors.lastname: Alber
authors.forename: Matthias
authors.initials: M
authors.lastname: Frick
authors.forename: Jozef
authors.initials: J
authors.lastname: Dulak
authors.forename: Otmar
authors.initials: O
authors.lastname: Pachinger
authors.forename: Franz
authors.initials: F
authors.lastname: Weidinger
mesh_terms: D000818:Animals; D001013:Aorta, Thoracic / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007249:Inflammation / Q000378:metabolism / Q000473:pathology; D008297:Male; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000187:drug effects; D020123:Sirolimus / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000378:metabolism; D017539:Tunica Intima / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D017540:Tunica Media / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D007166:Immunosuppressive Agents; D016328:NF-kappa B; D014409:Tumor Necrosis Factor-alpha; D020123:Sirolimus