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Low Molecular Weight Heparin Fragment Decreases Intracellular Calcium in Human Hepatocarcinoma Cells Under Calcium Overload
Ênio R. Vasques^*1, José Eduardo M. Cunha¹, Ana Maria M. Coelho¹, Sandra N. Sampietre¹, Emilio E. Abdo¹, Helena B. Nader², Ivarne S. Tersariol², Marcelo A. Lima², Luiz Augusto C. D'Albuquerque¹, Eleazar Chaib¹, Tiago Rodrigues^3
1GASTROENTEROLOGY, FMUSP, São Paulo, São Paulo, Brazil; ²PHARMACOLOGY, UNIFESP, SAO PAULO, Brazil; ³FEDERAL UNIVERSITY OF ABC, SANTO ANDRE, Brazil

Background: Liver cell damage secondary to Ischemia and reperfusion (I/R) that occurs in liver surgery and transplantation is associated to intracellular Calcium overload. It has been previously demonstrated that the sodium calcium exchanger (NCX), which regulates intracellular calcium extrusion, is affected by low molecular weight heparin fragments in rabbits endothelial aortic cells, decreasing intracellular calcium through the inhibition of the exchange inhibitory peptide (XIP). However, their action in human hepatocytes has not been previously investigated. Objective: To evaluate the effect of a low molecular weight heparin fragment, the trisulfated dissacharide (TD), on human hepatocytes under calcium overload. Material and Methods: HegG2 culture cells from Human Hepatocellular Carcinoma that express NCX protein were grown in Dulbecco’s modified Eagle’s medium for 24h and loaded with the fluorescent Ca2+ indicator fura-2/AM (4 μM). After twice cell washing, 1.0 mL of specific medium were add to the cell culture and the images acquired on a widefield microscope. Changes in cytosolic calcium levels were measured at 37C for 15 minutes. Thapsigargin 4 µM was used to promote a sustained raise in the cytosolic calcium level. TD 30 µM was added to the medium 90 seconds after Thapsigargin. Results: TD added to the medium after Thapsigargin decreased the intracellular calcium levels when compared to the control group (Figures 1 and 2). Conclusion: TD was able to decrease the intracellular calcium raise promoted by Thapsigargin in human hepatocarcinoma cells under calcium overload

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