Ann Surg Treat Res.  2015 Mar;88(3):152-159. 10.4174/astr.2015.88.3.152.

Effect of imatinib mesylate and rapamycin on the preformed intimal hyperplasia in rat carotid injury model

• Park YJ
• Min SK
• Min SI
• Kim SJ
• Ha J

• KMID: 2266873
• DOI: http://doi.org/10.4174/astr.2015.88.3.152

Abstract

PURPOSE
Intimal hyperplasia (IH) is the main cause of restenosis or occlusion after vascular procedures. Imatinib mesylate and rapamycin are known to prevent IH. The purpose of this study was to evaluate the effect of these drugs on the regression of preformed IH in rat carotid injury model.
METHODS
IH was established in rat carotid arteries using a balloon catheter. The drug effects were assessed in vitro on proliferation, migration, and apoptosis of vascular smooth muscle cells (VSMC) in the neointima. And in vivo studies were carried out in 4 groups: imatinib, rapamycin, combined, and no medication. After 2-week oral medication, morphometric analysis evaluated the number and density of neointimal cells, intima-to-media (I/M) ratio and cross-sectional area. Cell proliferation, apoptosis, and collagen changes were also investigated by immunohistochemical staining (IHCS).
RESULTS
Imatinib and rapamycin significantly inhibited VSMC proliferation and migration, and promoted apoptosis in vitro. In morphometric analysis, the number and density of neointimal cells decreased significantly in all medication groups compared with control group (P < 0.01). However, there was no significant difference in neointimal cross-sectional area and I/M ratio among groups. In IHCS, imatinib and rapamycin inhibited neointimal cell proliferation significantly. However, there was no significant change in cell apoptosis and collagen composition.
CONCLUSION
Combined treatment of with imatinib and rapamycin induced reduction of cell mass in preformed intimal hyperplasia, but failed to induce regression of intimal mass in this short-term medication study. Further studies will be needed with additional strategies of inducing lysis of the extracellular matrix.

Keyword

Imatinib; Sirolimus; Neointima; Hyperplasia; Regression

MeSH Terms

Animals
Apoptosis
Carotid Arteries
Catheters
Cell Proliferation
Collagen
Extracellular Matrix
Hyperplasia*
Mesylates*
Muscle, Smooth, Vascular
Neointima
Rats*
Sirolimus*
Collagen
Mesylates
Sirolimus

Figure

• 

  Fig. 1 Cross-sectional images of the injured carotid arteries serially at 3 (A), 7 (B), 14 (C), and 28 days (D) after injury (H&E, light microscopy, ×100). Intimal hyperplasia was developed progressively after the injury. The neointima was built up enough at 14 days after injury, comparable to that at 28 days.

• 

  Fig. 2 Cell proliferation study with bromodeoxyuridine assay. Vascular smooth muscle cells from the neointimal of the injured left common carotid artery were used. Cell proliferation significantly decreased along with the drug concentrations, especially in more than 10-7M of all medication groups (*P < 0.05 vs. control; **P < 0.01 vs. control).

• 

  Fig. 3 Cell migration study with wound scratch assay. Vascular smooth muscle cells migration activity was significantly inhibited in all medication groups, especially in more than 10-8M (*P < 0.05 vs. control; **P < 0.01 vs. control).

• 

  Fig. 4 Cell apoptosis by fluorescence-activated cell sorter (FACS) assay. Representative data of FACS analysis showed that apoptotic cells significantly increased in imatinib mesylate (B) and combined (D) groups. However, rapamycin (C) group showed no significant difference (***P < 0.001 vs. control [A]).

• 

  Fig. 5 Morphometric analysis. (A) Mean neointimal cell counts were assessed under the four random high-power fields (×400). Mean neointimal cell counts significantly decreased in all medication groups compared to control (*P < 0.05 vs. PBS, **P < 0.01 vs. PBS). (B) Neointimal cross-sectional area was not significantly different among the groups. (C) The ratio of intima and media thickness was not significantly different among the groups.

• 

  Fig. 6 Cell Proliferation Index. Immunohistochemical staining (×400) against proliferating cell nuclear antigen (PCNA) showed that PCNA-positive cells were significantly decreased in all medication groups compared to control, which represented that cell proliferation was inhibited by each drug medication (***P < 0.001 vs. control).

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