The Effects Of Calcium Antagonist Verapamil In The Murine Model Of Traumatic Hemorrhagic Shock With Endotoxemia

Objective: This study is designed to evaluate the effects of calcium antagonist verapamil in the murine model of traumatic hemorrhagic shock with endotoxemia and to investigate the changes of phosphorylated form of p38MAPK in macrophages.Methods: Part One: Fifty-four male Sprague Dawley rats were randomly assigned into three groups with nine in each. Heart Rate(HR) and mean arterial pressure(MAP) of the rats were measured from the beginning of the experiment. Plasma concentrations of TNF- α , IL-1 β , IL-10 were measured with ELISA in each group 90 min after LPS treatments. Peritoneal macrophages were collected at the same time, and from which cellular protein were extracted and analyzed by Western blot for the phosphorylated form of p38MAPK. Histological examination of lung and small intestine were performed at the points of 4 h and 24 h after LPS exposure.Part Two: Peritoneal macrophages were collected from twelve male Sprague Dawley rats and separated immediately in six-well plates with 5 × 10~6 cells per well. Cells were exposed to LPS(100ng/ml) in the presence of verapamil(50μmmol/l) or SB203580(10μmmol/l). TNF- α , IL-1 β and IL-10 expressions were measured by ELISA on cellular supernatants after 90 min of LPS treatments. Cellular protein were extracted and analyzed by Western blot for the phosphorylated form of p38MAPK.Results: Part One: Rats in the group of traumatic hemorrhagic shock with endotoxemia showed higher plasma concentrations of TNF- α , IL-1 β and IL-10 and a higher level of phosphorylated p38MAPK in the peritoneal macrophages when compared with the control group (p<0.01) . Rats in the group of Verapamil treatment(10mg/Kg) showed lower plasma concentrations of TNF- α , IL-1 β and higher plasma concentration of IL-10 when compared with Trauma group (p<0.01) . Verapamil had no significant effect in Trauma induced p38MAPK phosphorylation in macrophages (p>0.05). Rats pretreated with verapamil led to attenuated inflammatory histological changes in lung and small intestine when compared with Trauma group.