J Korean Med Sci.  2004 Jun;19(3):327-332. 10.3346/jkms.2004.19.3.327.

Expression of Cell Cycle Regulators During Smooth Muscle Cell Proliferation After Balloon Catheter Injury of Rat Artery

• Chung JK
• Lee T
• Jung IM
• Kim YK
• Min SK
• Suh JW
• Kim SJ

• KMID: 1786807
• DOI: http://doi.org/10.3346/jkms.2004.19.3.327

Abstract

Intimal hyperplasia is defined as the abnormal migration and proliferation of vascular smooth muscle cells (VSMCs) with deposition of extracellular matrix. However, the cell cycle regulatory mechanisms of injury-induced VSMC proliferation are largely unknown. To examine the expression kinetics of cell cycle regulatory factors which is known to be worked positively or negatively, we used rat balloon injury model. Marked induction of proliferating cell nuclear antigen (PCNA), G1/S cyclin-dependent kinase (cdk2), and its regulatory subunit (cyclin E) occurred between 1 and 3 days after balloon arterial injury, and this was sustained for up to 7 days and then declined. However, the induction of the negative regulators, p21 and p27, occurred between 3 and 5 days of injury, peaked after 7 and 14 days and was then sustained. VSMC proliferation after balloon catheter injury of the rat iliac artery is associated with coordinated expression of positive (cdk2, cyclin E and PCNA) and negative (p21, p27) regulators. Cell cycle regulators such as cdk2, cyclin E, p21, p27 may be suitable targets for the control of intimal hyperplasia.

Keyword

Endothelium, Vascular; Myocytes, Smooth Muscle; Tunica Intima; Cell Cycle Proteins; Rat

MeSH Terms

Animals
Arteries/*pathology
Balloon Dilatation/*adverse effects
Blotting, Western
CDC2-CDC28 Kinases/biosynthesis
Cell Cycle
Cell Cycle Proteins/biosynthesis
Cell Division
Cyclin E/biosynthesis
Cyclins/biosynthesis
Endothelium, Vascular/pathology
Extracellular Matrix/metabolism
Hyperplasia/pathology
Iliac Artery/pathology
Immunohistochemistry
Male
Myocytes, Smooth Muscle/*cytology
Proliferating Cell Nuclear Antigen/biosynthesis
Rats
Rats, Sprague-Dawley
Support, Non-U.S. Gov't
Time Factors
Tumor Suppressor Proteins/biosynthesis

Figure

• 

  Fig. 1 Microscopic findings: photomicrograph of cross section of the injured rat arteries (H&E): Normal iliac artery (control group) with single layer of endothelium in intima and prominent intima was seen from 3 days, marked thickened in 7 days, and sustained thereafter. I, indicate intima and M, media. Black arrows point to the internal elastic lamina. Magnification ×200.

• 

  Fig. 2 Sequential change in expression of cell cycle regulators (cdk 2, cyclin E, PCNA, p21, p27) by Western blot analysis after balloon catheter injury of rat iliac artery. Marked induction of cdk2, cyclin E, PCNA occurs between 1 and 3 days after balloon arterial injury, and is peaked in 5 or 7 days and then declined. But induction of negative regulators, p21 and p27 occurs between 3 and 5 days after injury, and is peaked 7 or 14 days and then sustained.

• 

  Fig. 3 Immunohistochemical staining of cdk2 after balloon catheter injury of rat iliac artery. Note that cdk2 expression in the medial cells is upregulated within 3 days (C) and then declined thereafter (D, G). At later time points, cdk2 expression is detected throughout emerging intimal lesion (D-E). White arrows point to the internal elastic lamina. Magnification ×200.

• 

  Fig. 4 Immunohistochemical staining of cdk2 after balloon catheter injury of rat iliac artery. Cyclin E expression in the media is noted early at 3 days and 5 days (C, D) and then declined thereafter (E, F). At later time points, cyclin E expression is noticed throughout emerging intimal lesion (D-E). White arrows point to the internal elastic lamina. Magnification ×200.

• 

  Fig. 5 Immunohistochemical staining of p21 after balloon catheter injury of rat iliac artery. Note that p21 expression in the medial cells is upregulated at 5 days (D) and then sustained thereafter (E, F). At later time points, cdk2 expression is detected throughout emerging intimal lesion (E-F). White arrows point to the internal elastic lamina. Magnification ×200.

• 

  Fig. 6 Immunohistochemical staining of p27 after balloon catheter injury of rat iliac artery. Note that p27 expression in the medial cells is upregulated at 3 days and 5 days (C, D) and then sustained thereafter (E, G). At later time points, p27 expression is detected throughout emerging intimal lesion (D, G). White arrows point to the internal elastic lamina. Magnification ×200.

• 

  Fig. 7 Immunohistochemical staining of PCNA after balloon catheter injury of rat iliac artery. PCNA expression is noted in the medial at 3 days (C) and then sustained at 5 and 7 days (D, G) and then declined (F, G). PCNA expression is detected throughout emerging intimal lesion (D-E). White arrows point to the internal elastic lamina. Magnification ×200.

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