Prolonged increases in vein wall tension increase matrix metalloproteinases and decrease constriction in rat vena cava: Potential implications in varicose.

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Presentation transcript:

Prolonged increases in vein wall tension increase matrix metalloproteinases and decrease constriction in rat vena cava: Potential implications in varicose veins Joseph D. Raffetto, MD, Xiaoying Qiao, MD, PhD, Vera V. Koledova, MD, PhD, Raouf A. Khalil, MD, PhD Journal of Vascular Surgery Volume 48, Issue 2, Pages 447-456 (August 2008) DOI: 10.1016/j.jvs.2008.03.004 Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 1 Tension-potassium chloride (KCl) contraction relationship. Inferior vena cava segments were subjected to increasing basal tensions (0.0625 to 3 g) for 30 minutes, then stimulated with 96 mmol/L KCl. KCl contraction was not significantly different at 0.5 to 2 g basal tension. Further increase in wall tension to 3 g was associated with significant decrease in contraction (P < .01). Data points represent means ± SEM of measurements from eight experiments. Journal of Vascular Surgery 2008 48, 447-456DOI: (10.1016/j.jvs.2008.03.004) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 2 Effect of increases in basal tension on phenylephrine (Phe)- and angiotensin II (AngII)-induced contraction. Inferior vena cava segments were subjected to 0.5 g tension for 1 hour or 2 g tension for 24 hours in the absence or presence of tissue inhibitor of metalloproteinase-1 (TIMP-1). The tissues were stimulated with Phe (10−5 mol/L) (A) or AngII (10−6 mol/L) (B) and the contractile response was recorded. Data points represent means ± SEM of measurements from four experiments. *Significantly different (P < .05) from contraction at 0.5 g for 1 hour basal tension. §Significantly different (P < .05) from contraction at 2 g for 24 hours basal tension. Journal of Vascular Surgery 2008 48, 447-456DOI: (10.1016/j.jvs.2008.03.004) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 3 Effect of increases in basal tension on KCl-induced contraction. Inferior vena cava segments were subjected to 0.5 g tension for 1 hour, or 2 g tension for 24 hours in the absence or presence of tissue inhibitor of metalloproteinase-1 (TIMP-1). The tissues were stimulated with KCl (96 mmol/L) and the contractile response was recorded. Data points represent the means ± SEM of measurements from four experiments. *Significantly different (P < .05) from KCl contraction at 0.5 g 1 hour basal tension. Journal of Vascular Surgery 2008 48, 447-456DOI: (10.1016/j.jvs.2008.03.004) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 4 Immunoblot analysis for matrix metalloproteinase (MMPs) in inferior vena cava (IVC). IVC segments were subjected to 0.5 g tension for 1 hour or 2 g tension for 24 hours. The tissues were rapidly frozen and prepared for Western blot analysis using specific anti MMP-2 (1:500) and anti MMP-9 antibody (1:200). The optical density of the immunoreactive bands was normalized to the house-keeping protein actin. Data points represent means ± SEM of eight to 15 measurements. *Significantly different (P < .05). Journal of Vascular Surgery 2008 48, 447-456DOI: (10.1016/j.jvs.2008.03.004) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 5 Correlation between matrix metalloproteinase (MMPs) expression and phenylephrine (Phe) and KCl contractile response. Inferior vena cava segments were subjected to 0.5 g tension for 1 hour or 2 g tension for 24 hours. After eliciting the contractile response to KCl (96 mmol/L) and Phe (10−5 mol/L), the vein segments were frozen and prepared for Western blot analysis using anti MMP-2 (1:500) and anti MMP-9 antibody (1:200). The optical density of the immunoreactive bands was normalized to actin. The relation between the amount of MMP-2 (A) or MMP-9 (B) and the contractile response to KCl and Phe was then constructed. Data points represent 15 measurements for MMP-2 and eight measurements for MMP-9. Journal of Vascular Surgery 2008 48, 447-456DOI: (10.1016/j.jvs.2008.03.004) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 6 Immunohistochemistry for matrix metalloproteinase (MMPs) in inferior vena cava (IVC). IVC segments at rest or subjected to 2 g tension for 24 hours were rapidly frozen, and cryosections were prepared for immunohistochemical staining using anti MMP-2 or MMP-9 antibody (1:500). The amount of brown positive staining in the tunica intima (I), media (M), and adventitia (A) was measured. Hematoxylin and eosin staining was performed to test for integrity of the vessel wall. Data represent measurements from four to five pictures of tissue sections. Total magnification 400. Journal of Vascular Surgery 2008 48, 447-456DOI: (10.1016/j.jvs.2008.03.004) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions

Fig 7 Effects of matrix metalloproteinase (MMPs) on phenylephrine (Phe) contraction of inferior vena cava (IVC) segments. IVC segments were subjected to 0.5 g tension for 1 hour and either nontreated or pretreated with TIMP-1. The tissues were stimulated with Phe (10−5 mol/L). When the Phe contraction reached a steady state, the tissues were treated with MMP-2 or MMP-9 (1 μg/ml) and the continuous trace of the effect of MMP on vein contraction was recorded for at least 30 minutes. To compare quantitatively the magnitude of Phe contraction in MMP and TIMP-1 treated and nontreated veins, the contraction trace was analyzed for the magnitude of the response at selected time points at 5-minute intervals for 30 minutes. Data points represent means ± SEM of measurements from five experiments. *Significantly different (P < .05) from control. Journal of Vascular Surgery 2008 48, 447-456DOI: (10.1016/j.jvs.2008.03.004) Copyright © 2008 The Society for Vascular Surgery Terms and Conditions