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Changes in the level of cytosolic calcium, nitric oxide and nitric oxide synthase activity during platelet aggregation: an in vitro study in platelets from normal subjects and those with cirrhosis

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Abstract

Variceal bleeding due to abnormal platelet function is a well-known complication of cirrhosis. Nitric oxide-related stress has been implicated in the pathogenesis of liver cirrhosis. In the present investigation, we evaluated the level of platelet aggregation and concomitant changes in the level of platelet cytosolic calcium (Ca2+), nitric oxide (NO) and NO synthase (NOS) activity in liver cirrhosis. The aim of the present study was to investigate whether the production of NO by NOS and level of cytosolic Ca2+ influence the aggregation of platelets in patients with cirrhosis of the liver. Agonist-induced aggregation and the simultaneous changes in the level of cytosolic Ca2+, NO and NOS were monitored in platelets of patients with cirrhosis. Platelet aggregation was also measured in the presence of the eNOS inhibitor, diphenylene iodinium chloride (DIC). The level of agonist-induced platelet aggregation was significantly low in the platelets of patients with cirrhosis compared with that in platelets from normal subjects. During the course of platelet aggregation, concomitant elevation in the level of cytosolic Ca2+ was observed in normal samples, whereas the elevation was not significant in platelets of patients with cirrhosis. A parallel increase was observed in the levels of NO and NOS activity. In the presence of the eNOS inhibitor, platelet aggregation was enhanced and accompanied by an elevated calcium level. The inhibition of platelet aggregation in liver cirrhosis might be partly due to greater NO formation by eNOS. Defective Ca2+ release from the internal stores to the cytosol may account for inhibition of aggregation of platelets in cirrhosis. The NO-related defective aggregation of platelets in patients with cirrhosis found in our study is of clinical importance, and the underlying mechanism of such changes suggests a possible therapeutic strategy with cell-specific NO blockers.

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Abbreviations

ADP:

adenosine diphosphate

ATPase:

adenosine triphosphatase

BT:

bleeding time

cGMP:

cyclic guanosine monophosphate

cNOS:

constitutive NOS

DIC:

diphenylene iodinium chloride

eNOS:

endothelial NO synthase

iNOS:

inducible NOS

NO:

nitric oxide

NOS:

No synthase

nNOS:

neuronal NO synthase

PRP:

platelet-rich plasma

RBC:

red blood cell

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Authors and Affiliations

  1. Department of Biochemistry, Bharathi Women’s College, Chennai, 600 108, India

    Sam Annie-Jeyachristy & Arumugam Geetha

  2. Department of Surgical Gastroenterology and Proctology, Stanley Medical College and Hospital, Chennai, 600 108, India

    Rajagopal Surendran

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  1. Sam Annie-Jeyachristy
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  2. Arumugam Geetha
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  3. Rajagopal Surendran
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Correspondence to Arumugam Geetha.

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Annie-Jeyachristy, S., Geetha, A. & Surendran, R. Changes in the level of cytosolic calcium, nitric oxide and nitric oxide synthase activity during platelet aggregation: an in vitro study in platelets from normal subjects and those with cirrhosis. J Biosci 33, 45–53 (2008). https://doi.org/10.1007/s12038-008-0020-0

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