The Effect Of The Argatroban On The Mouse Thrombopenia Induced By LPS

ObjectiveThrombocytopenia (TCP),which is a common complication in Intensive Care Unit(ICU),is one of the dangerous sign towards the patient with severe sepsis.It is considered Systemic Inflammatory Response Syndrome(SIRS) resulted from infection is the important reason to TCP.But the definite mechanism of TCP,which is induced by LPS,have not to be explicit.The mechanism maybe contain diffuse intravascular coagulation(DIC),immune mediated platelet destruction, hemophagocytic syndrome and so on.The experiment by establishing LPS mice induced thrombocytopenia model, to observe the influence of direct thrombin inhibitors argatroban on the mouse with TCP, judge LPS induction thrombocytopenia happens.MethodsThis experiment was divided into two parts.The first part established C57BL/6mice thrombocytopenia model which was induced by LPS. The SPF C57BL/6mouse were all male and4-6weeks old.The body weight were18-22g.75mouses were randomly divided into control group (the C group)、low dose LPS group(the L group),middle dose LPS group(the M group), high dose LPS group(the H group)and Extremely high doses LPS group(the S group), Each group of15only, Group C gave intraperitoneal injection of saline0.005ml/g; L、H、M、S group mouse were respectively injected LPS0.05mg/kg0.5mg/kg5mg/kg LPS50mg/kg by intraperitoneal.Wached and recorded the situation and the dead time of mouse, According to the point took blood of inner canthus and counted Platelet numerical changes.The second part observed the influence of direct thrombin inhibitors argatroban on the C57BL/6mouse with TCP,which is induced by LPS.The experiment used SPF male C57BL/6mouse.The mouse were6-8weeks old.The body weight were23-25g.72mouse were randomly divided into control group (the C group),low dose LPS group(the LL group),high dose LPS group(the HL group),low dose argatroban and low dose LPS group(the LALL group),high dose argatroban and low dose LPS group(the HALL group),low dose argatroban and high dose LPS(the LAHL group),high dose argatroban and high dose LPS group(the HAHL group). Each group of15only.All group measure base vasule before administering drug through endocanthion.4hours after administering drug measured peripheral blood platelets count(PC)、MPV and WBC.The rest of whole blood was centrifuged,1000*g,15min.then the blood plasm was extracted.We used the plasm to measure TAT、FDP、D-Dimer and TPO.The third step used intestine to HE staining,then observed the the change of histopathology by light microscope.ResultsThe first part in the experiment group C all time points platelet count was no statistical difference in mouse (P>0.05), L、M、H、S group injected mouse with LPS2h, platelet count dropped significantly comparied the basic value (all p<0.01),the L group PC down to a minimum at8h (p<0.01),24h began to have a modest recovery,to48h restore to the base value level; M group and H group mouse PC dropped to the lowest at24h (p<0.01),48h resumed, M group recovered to the base value level at72h, PC of the S group dropped to the lowest at72h (p<0.01)The second part of the experimental results as flows:compared with group C,the PC and WBC of the LL group and the HL group C were low.The PC of the HL group was decreased more than the LL group (p<0.01); The plasma levels of the TAT in the LL group and the HL group were lower than the group C (p<0.05), The plasma levels of the D-Dimer, FDP and TPO in the LL group and the HL group had no difference with the group C (p>0.05); The PC and WBC and the plasma levels of the TAT, D-Dimer, FDP and TPO in the LA group and the HA group had no significant difference with the group C (p>0.05); The PC and WBC and the plasma levels of the TAT in the LALL group and the HALL group were lower than the group C (P<0.05).But the plasma levels of the D-Dimer, FDP and TPO had no significant difference with the group C (p>0.05). The PC and WBC and the plasma levels of the TAT in the LAHL group and the HAHL group were lower than the group C (P<0.05). The plasma levels of the D-Dimer, FDP and TPO had no significant difference with the group C too.(p>0.05).The pathological grades of intestine in the LL group、HL group、LALL group、HALL group、LAHL group and HAHL group were higer than in the C group,but LA group and HA group were no statistically significant with group C.ConclusionThe blood platelet of the C57BL/6mouse were sensitive to endotoxin.TheTCP could be induced after intraperitoneal injection of LPS (0.05mg/kg)2hours.And with the dose and time of LPS increased,the toxicity symptom and the degree of the thrombocytopenia were more serious. The mortality rate of mouse were73.3%within injectioned LPS (50mg/kg)72hours. The declined percent of the peripheral blood platelet count were60%after Injectioned LPS4hours, and the white blood count also reduced significantly.the more the dose of LPS injected, the more obvious the declined degree of the platelet count were. Argatroban have no effect on the platelet count and the white blood cell count and also could induced the happening of TCP. LPS of the induction The thrombocytopenia of the mouse induced by LPS could be concerned with platelet production reduce and dramge increase.