The Change Of Dermatan Sulfate Concentration In Preeclampsia Before And After Delivery

PrefaceHypertensive disorder complicating pregnancy is one common complication of obsterics and weapon the health and life of newborn and mothers . The pathologic manifestation of serious preeclampsia imply disseminated intravascular coagulation (DIC) is the important procedure of the disease . Some study imply that normal late pregnant women is PTS , moreover the preeclampsia have the obvious PTS . We should take the efficient and reasonable measures to prevent and treat the chronic DIC of preeclampsia . This can prevent the severe complications , such as placen-tal abruption ,placental infarction , FGR , habitual abortion , fetal death and premature delivery .Dermatan sulfate (DS) belongs to the GAGS and widely distribute in the connective tissue of mammalian. It is a new antithrombotics and act through HC— Ⅱ ,not AT— Ⅲ ,DS can complete one third antithrombase activity . DS is the specific activator of HC— Ⅱ . It has wide perspective as a new antithrombotics . Some literature imply that DS has obvious cooperation in anticoagulant action with UFH . And the anticoagulant activity increase with the concentration of UFH when the concentration of DS is fixed .Some literature report that there are some concentration of dermatan sulfate . The study design detect the anticoagulant activity of DS in preeclampsia to observe- its level . This can supply some base for DS treatment in preeclampsia , and direct the anticoagulant treatment for pre-eclampsia .Materials and MethodStudy objectiveControl group :Normal pregnant women ,20 cases (team I );experiment group:light preeclampsia late term pregnant women 20 cases (team II );serious preeclampsia 20 cases (team HI) ? all the above patients are in hospital in the second affiliated hospital of china medical university . The prognostic principle of preeclampsia accord to editor Lejie Obsterics and Gynecology (2003) . Study objectives rule out liver dysfunction , and they didn't use the anticoagulant drugs , and they hadn't any other system disease. There is no statistic difference in age and gestational weeks. Informed consent from the patients for peripheral blood sampling was obtained.Materials and Method1. Materials1.1 Reagent -.bentonite (Sigma) , purified human ctthrombin(Haema-tologic Technologies) ,CBS34. 47(Stago) , purifed human heparin cofactor II (Haematologic Technologies ), dermatan sulfate (Sigma), chondroitin sulfate ABC lyase (Stago) , 50% glacial acetic acid (The first chemicals factory of SHENYANG).1. 2Equipment : electronic scale,stirrer bar, test tube, sample injector, incubator,centrifuger , spectrophotometer (405nm)2. Method2. lPlasma samples: Paired blood samples were obtained , with informed consent, from consecutive women with 20 normal pregnancies,20 light preeclampsia and 20 serious preeclampsia patients within 24 h before elective caesarean section and 3 —5 d postpartum. Blood was collected into 3. 8% natrium citricum (9:1 blood/ natrium citricum)and plasma ob-taind by centrifugation at 2000g for 15 minutes to get supernatant , — 70°C freezed to storage. These are needed to be done in 6 hours.2. 2The assay of DS anticoagulant activity :2. 2. lSample assay : By chromogenic substrate method . To get plasma sample , add the same volume 50g/L bentonite , strong mixed , After centrifugation at 2000g/min for 15 minutes , the supernatant was collected and a lOOjA sample brought 37°C. Purified human heparin cofac-tor II IOOjjiK 0. 026u/ml ) was added and incubated at 37°C for 2 min . Thrombin ,100jzl(6NIHU/ml) ,was added and incubated for 2 min . The synthetic thrombin substrate CBS34. 47,100^1(0. 16ptmol/ml) , was added and incubated for 2 min . The reaction was stopped with 100{A of 50% acetic acid and the absorbance was read at 405 nm . This was compared to a standard curve generated using NS to which varying amounts (0. l^g/ ml, 0. 5jug/ml, 1. 0/zg /ml, 1. 5/zg/ml, 2. 0/ig/ml) of DS had been added and the bentonite step was eliminated . All positive results were confirmed by reaction of bentonite—treated plasma with chondroitin ABC lyase (final concentration 0. 33U/ml) for 1 h at 37°C before assay . This was to confirm that any activity present was due to DS rather than heparin or hepa-ran sulfate , since these are also active in this assay system.3. Statistical treatment: We take SPSS11. 5 to analysis the OD to get the results . We take the ANOVA interclass and paired t—test in—class. P0. 05). The plasma levels of anticoagulant dermatan sulfate activity in team El , team H and team I all restored to normal after delivery(p>0. 05). The plasma levels of anticoagulant dermatan sulfate activity of all the team after delivery decreased more significantly than before delivery (p