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Activated Rho/Rho kinase and modified calcium sensitivity in cryopreserved human saphenous veins
Else Müller-Schweinitzer1, David C Reineke, Erika Glusa
1Department of Cardiac Surgery, University Hospital, CH-4031 Basel, Switzerland. dremueller36@bluewin.ch
Cryobiology
|July 8, 2008
Summary
Cryopreservation of human saphenous veins activates Rho kinase signaling pathways and alters intracellular calcium levels. This impacts vascular smooth muscle cell function and responses to contractile agents.
Area of Science:
- Vascular Biology
- Cryobiology
- Cell Signaling
Background:
- Cryopreservation of human internal mammary arteries activates protein kinase C and enhances intracellular calcium ([Ca2+]i).
- Cryoinjury in human saphenous veins (HSV) is associated with Rho/Rho kinase pathway activation and elevated [Ca2+]i.
Purpose of the Study:
- To investigate the effects of cryopreservation on human saphenous veins (HSV).
- To determine the role of Rho kinase signaling pathways and intracellular calcium in cryoinjury of HSV.
Main Methods:
- Human saphenous veins (HSV) were studied in vitro, either fresh or cryopreserved at -196°C.
- Cryopreserved HSV were assessed for contractile responses to agonists and CaCl2.
- Intracellular calcium ([Ca2+]i) was measured using Fluo-3AM fluorescence.
- Specific inhibitors and activators of ion channels and signaling pathways were employed.
Main Results:
- Cryostorage diminished contractile responses to receptor-mediated agonists (noradrenaline, 5-HT, endothelin-1) by up to 30% and KCl by 50%.
- The Ca2+ channel activator (+)-(S) SDZ 202-791 was 10-fold less effective in cryopreserved HSV.
- Rho kinase blockade by HA-1077 significantly attenuated contractile responses in cryopreserved HSV.
- [Ca2+]i measurements confirmed functional effects, showing altered Ca2+ handling post-cryopreservation.
Conclusions:
- Cryopreservation modifies intracellular calcium ([Ca2+]i) in venous smooth muscle cells.
- These modifications occur via depolarization-induced Ca2+ influx changes.
- Activation of Rho kinase signaling pathways is a key mechanism in cryoinjury of HSV.
