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Huh, Choi, Kim, and Kim: The Expression of Matrix Metalloproteinases and Tissue Inhibitors of Metalloproteinases in the Wall of Great Saphenous Vein in Patients with Varicose Veins
Original Article
Journal of the Korean Surgical Society

Journal of the Korean Surgical Society 2010; 79(Suppl 1): S16-S25.

Published online: 31 December 2010

DOI: https://doi.org/10.4174/jkss.2010.79.Suppl1.S16

The Expression of Matrix Metalloproteinases and Tissue Inhibitors of Metalloproteinases in the Wall of Great Saphenous Vein in Patients with Varicose Veins

Seung Huh, M.D., Hyang Hee Choi, M.D.1, Hyung-kee Kim, M.D., Sang Joon Kim, M.D.2

Division of Transplantation and Vascular Surgery, Department of Surgery, Kyungpook National University School of Medicine, Daegu, Korea.

1Department of Surgery, Pohang St. Mary's Hospital, Pohang, Korea.

2Department of Surgery, Seoul National University College of Medicine, Seoul, Korea.

Corresponding author (shuh@knu.ac.kr)

Received 27 May 2010       Accepted 28 September 2010

Copyright © 2010 The Korean Surgical Society

Abstract

Purpose

Although varicose veins are very common in adults, the mechanism of the disease has not been established. Degradation of the extracellular matrix is regulated by various matrix metallopreteinases (MMPs) and their inhibitors tissue inhibitor of metallaproteinase (TIMPs). This study was performed to analyse the relationship between venous wall degeneration and expression of these matrix proteinases.

Methods

Twelve great saphenous vein (GSV) segments from 7 patients without varicose veins (control) and 86 GSV segments from 18 patients (22 limbs) with varicose veins (C2,4,5EPASPR) were used for this study. Light microscopic examination was used in the evaluation of vein wall degeneration, immunohistochemistry and Western blotting for the expression of MMPs (MMP-1, MMP-2, MMP-9 and MMP-13) and TIMPs (TIMP-1 and TIMP-2), and zymography for gelatinolytic activity of MMP-2 and MMP-9 were performed.

Results

MMP-9 was more strongly expressed in the vein wall of both control and patient groups, especially in the endothelial cells and medial muscle layers and TIMP-2 followed. The expression of MMP-9 was closely related to the degree of venous wall degeneration. Activated MMP-2 and MMP-9 were observed in both groups and expressed more in the proximal GSV of the patients. In the Western blotting, the expression of MMP-9 and TIMP-1 were significantly higher than other MMPs and TIMP-2 in the patients with varicose veins.

Conclusion

MMP-9 is much more expressed in the wall of degenerative veins. This matrix-degrading enzyme may play an important role in the degeneration of venous wall followed by its remodeling.

Keywords: Varicose veins, Matrix metalloproteinase, Tissue inhibitor of metalloproteinase

Figures and Tables

Fig. 1
These photos show histological findings of the proximal segment of the great saphenous vein without axial reflux obtained from a 49-year-old male (control) who underwent living-related liver transplantation. The intimal (black arrow) and medial muscle layers are relatively well preserved (A) (Masson's trichrome stain, ×100). Also the internal elastic lamina (black arrow) and black-colored elastin fibers in medial muscle layers are not disrupted (B) (Verhoff's elastic fiber stain, ×100).
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Fig. 2
These photos show histochemical findings of the proximal segment of diseased saphenous vein obtained from a 57-year-old male with varicose vein. There are abundant intimal hyperplasia (black arrow, ↔) and disorganized mid and outer medial muscle layers (black arrow, ←) with increased blue-colored collagen fibers and fibrosis (A) (Masson's trichrome stain, ×100). Also the internal elastic lamina is disrupted (black arrow) and black-colored degenerative elastin fibers in medial muscle layers are fragmented (B) (Verhoff's elastic fiber stain, ×100). The matrix metalloproteinase-1 (MMP-1) is not expressed (C) (×100), whereas MMP-9 is strongly expressed (brown color) through endothelial cells (single black arrow), mid-outer medial muscle layers (double black arrows) and vasavasorum (black arrow head) (D) (×100).
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Fig. 3
Degree of degeneration in vein wall according to sites and layers of patients with varicose veins.The changes of medial layers are more prominent than intimal layers regardless of the sites of vein segments (P<0.05).
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Fig. 4
The degree of expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) according to the sites and the layers of the patients with varicose veins.The expression of MMP-9 is much higher than other MMPs and TIMPs regardless of the sites of vein segments (P<0.05).
jkss-79-S16-g004
Fig. 5
This photo shows zymographic activity of matrix metalloproteinase- 2 (MMP-2) and MMP-9 in control and varicose veins. P = Proximal; H = Hunter; D = Dodd; B = Boyd.
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Fig. 6
This photo shows Western blotting expression of matrix metalloproteinase-1 (MMP-1), MMP-2, MMP-9, MMP-13, tissue inhibitors of metalloproteinase-1 (TIMP-1), and TIMP-2 protein in control and varicose veins. P = proximal; H = Hunter; D = Dodd; B = Boyd.
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Table 1
Comparison between the clinical category and the number of severely dilated venous valves of the patients with varicose veins (n=22 limbs)
jkss-79-S16-i001

*CEAP (C = clinical; E = etiological; A = anatomical; P = pathophysiological), represented by an ad hoc committee of the American Venous Forum, 1994.

Table 2
The zymographic activities of the controls and the patients
jkss-79-S16-i002

*MMP = matrix metalloproteinases; †The activities of proximal site are higher than those of Hunter and Dodd sites (P<0.05).

Table 3
The results of Western blotting in the patients with varicose veins
jkss-79-S16-i003

*MMP = matrix metalloproteinases; †TIMP = tissue inhibitor of metalloproteinase; ‡P =0.045; §The total amounts of MMP-9 and TIMP-1 are more than those of other MMPs and TIMP-2 (P<0.05).

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