Abstract
Background: Beside its anti-proliferative, anti-hypertensive and anti-inflammatory effects, heparin has shown the apoptotic effect in lymphoblasts. In the study, it is aimed to show the apoptotic effect of heparin in lymphoblasts with measuring intracellular calcium and using DNA analysis by flow cytometry, in vitro. Methods: Twenty-three newly diagnosed acute lymphoblastic leukemia patients were included in the study. We added 10 and 20 U/ml heparin into the seperated lymphoblast samples and determined the percentages of apoptosis and intracellular Ca++ levels at 0, 1 and 2 hours by flow cytometry, in vitro. Results: The apoptotic effect on the lymphoblasts were established in 10 and 20 U/ml heparin concentrations at 0, 1 and 2 hours (p=0.005). The apoptotic effect of heparin in lymphoblasts was higher at the first hour than those at 0 and 2 hours in 10 and 20 U/ml heparin concentrations (p=0.005). The highest apoptosis was determined in 20 U/ml heparin concentration at the first hour. Statistically significant increase in intracellular Ca++ levels were determined in 10 and 20 U/ml heparin concentrations at 1 and 2 hours (p=0.005). In 10 and 20 U/ml heparin concentrations, intracellular Ca++ levels were significantly higher at the first hour than 0 and 2 hours (p=0.005). The highest intracellular Ca++ concentration was determined in 20 U/ml heparin concentration at the first hour. Conclusion: Heparin induces apoptosis in lymphoblasts and intracellular Ca++ levels of the lymphoblasts synchronously increase with apoptosis. The increase of intracellular Ca++ level supports a concept that the mitochondria plays a role heparin-induced apoptosis in lymphoblasts.
Highlights
Beside its anticoagulant, anti-inflammatory, antihypertensive and antiproliferative effects [1,2,3], heparin has the apoptotic effect in lymphoblasts, in vitro [4,5,6]
Lymphoblasts were incubated with 0, 5, 10 and 20 U/ml concentrations of heparin at 0, 1 and 2 hours and the highest apoptosis was determined in 20 U/ml heparin concentration at first hour [4,5,6]
We aimed to measure the intracellular Ca++ levels in lymphoblasts incubated with heparin and to indicate mitochondria-mediated apoptosis of heparin by flow cytometry (FCM)
Summary
Anti-inflammatory, antihypertensive and antiproliferative effects [1,2,3], heparin has the apoptotic effect in lymphoblasts, in vitro [4,5,6]. Manaster et al [7] claimed that 50, 100 and 200 U/ml concentrations of heparin induce apoptosis in human peripheral blood neutrophils, in vitro. The expression of fas, the activities of caspase-3 and -8 increased and the expression of bcl decreased when lymphoblasts were incubated with heparin in 10 and 20 U/ml concentrations at 1 and 2 hours. We first reported the apoptotic effect of heparin in lymphoblasts and stated that heparin induces apoptosis via the activation of the extrinsic pathway [5,6]. We claimed that heparin induces intrinsic apoptosis of lymphoblasts by the activation of caspase-9 and Cytochrome C (Cyt C) [8]. It is aimed to show the apoptotic effect of heparin in lymphoblasts with measuring intracellular calcium and using DNA analysis by flow cytometry, in vitro
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