Effect And Mechanism Of The Blood Plasm Calcium Concentration On Inflammation Mediators In Maintenance Hemodialysis Patients

Chapter 1EFFECTS OF THE BLOOD PLASM CALCIUM CONCENTRATION ON INFLAMMATION MEDIATORS AND PULSE WAVE VELOCITY IN MAINTENANCE HEMODIALYSIS PATIENTSObjectivePulse wave velocity is a earlier index of atherosclerosis level and can reflect damage of all kinds of risk factors on blood vessel. It conduced to understand risk factors of cardiovascular complication and prevent and cure these complications for investigating the effects of blood plasm calcium concentration on inflammation mediators and pulse wave velocity in maintenance hemodialysis patientsMethods1 Sixty three maintenance hemodialysis patients were divided into two groups(A group n=31 and B group n=32). The blood plasm calcium concentrations of A group were in normal range.The blood plasm of 3 mL venous blood,anticoagulation by ethylene diamine tetraacetic acid,centrifugation 10 minutes by 1500 rpm,were preserved in -20℃refrigerator for measuring.2 Results of hepatic function,renal function,blood fat and the blood plasm calcium density were from our biochemistry compartment,C reactive protein(CRP),interleukin-6(IL-6),total Homocysteine(tHcy),parathyroid hormone(iPTH) were measured by enzyme-linked immunosorbent assay.3 carotid-femoral pulse wave velocity(CF-PWV) were detected by PWV analysator.4 SPSS 13.0 statistical package were utilized for handling data,measurement data were expressed as arithmetic mean and standard deviation( x±s).The comparisons between the stydy groups were performed with chi square test,t test,linear correlation analysis,multiple linear regression analysis and multiple logistic regression analysis.Results1 The levels of CRP, IL-6,tHcy ,iPTH and CF-PWV in A group were lower than those of B group(p﹤0.05);2 There was a negative correlation among the blood calcium concentration and CRP and IL-6 and iPTH and tHcy and CF-PWV in 63 patients by linear correlation analysis; 3 By multiple linear regression analysis,levels of blood calcium concentration and dialysis time correlated with levels of CRP and tHcy , dialysis time correlated with levels of IL-6;4 The blood calcium concentration,dialysis time,levels of CRP and tHcy correlated with increase of CF-PWV by multiple linear regression analysis;5 Tthe blood plasm calcium concentration,dialysis time and levels of tHcy correlated with increase of CF-PWV by multiple logistic regression.Conclusions1 There is a significant negative correlation among the blood plasm calcium concentration and CRP and tHcy;2 In addition to dialysis time, blood plasm calcium concentration and tHcy are independent risk factors of increase of CF-PWV in maintenance hemodialysis.chapter 2EFFECT OF DIFFERENT CONCENTRATION CALCIUM CHLORIDE ON PERIPHERAL BLOOD MONONUCLEAR CELL INFLAMMATION MEDIATORS,PKC AND NF-κB IN MHD PATIENTS ObjectiveAccording to prophase clinical observe founds,continually investigating the effect of calcium chloride of different concentration on peripheral blood inflammation mediators,PKC and NF-κB in MHD patients .Methods1 Peripheral blood mononuclear cells,from 6 healthy volunteers and 6 MHD patients were obtained after centrifugalization,5×105PBMCs/well ,seeded in 96 wells cell culture plate were divided into 5 groups for cultivating 12 hours.A group:normal PBMC,B group:patient PBMC,C group: patient PBMC+1.0mmol/L Cacl2,D group: patient PBMC+1.25mmol/L Cacl2,E Group: patient PBMC+1.5mmol/L Cacl2;2 Cell morphologic changes before and after culture were observed; CRP and IL-6 of culture supernatant were detected by ELISA; calcium ion concentration within cells were measured by Fluo-3 AM;3 PKC and NF-κB within cells were detected by western blot.Results1 Cell appearance and amount before and after 12 hours culture didn,t show marked change in every groups;2 Discrepancies of calcium ion concentration within cells after cultivating 12 hours between every groups were not significant; 3 There were not obvious difference about levels of CRP and IL-6 after 12 hours culture between A and B groups,so were leves of PKC and NF-κB;4 The levels of CRP and IL-6 in C group after 12 hours culture were lower than E group;so were levels of PKC and NF-κB.Conclusions1 Calcium chloride of three kind concentration didn't affect form and amount of PBMC of MHD patients after cultivating 12 hours ;2 Discrepancies were not significant about effects of Calcium chloride of three kind concentrations on calcium ion concentration within PBMCs of MHD patients after cultivating 12 hours ;3 There were not obvious difference about levels of CRP and IL-6 from PBMC between healthy volunteers and MHD patients ;levels of PKC and NF-κB within PBMC in healthy volunteers were not significant,compared with MHD patients after cultivating 12 hours ;4 Compared with 1.5mmol/L calcium chloride,1.0mmol/L calcium chloride significantly inhibited CRP and IL-6 production of PBMC,reduced levels of PKC and NF-κB within PBMC in MHD patients. Chapter 3SYNERGETIC EFFECT OF LIPOPOLYSACCHARIDE AND CALCIUM CHLORIDE ON PERIPHERAL BLOOD MONONUCLEAR CELL INFLAMMATION MEDIATORS ,PKC AND NF-κB IN MHD PATIENTSObjectiveConsidering the uremic medium of MHD patients,for instance,PBMC were stimulated by lipopolysaccharide ,advanced glycosylation ends and so on.for this reason,to explore the synergetic effect of lipopolysaccharide and calcium chloride on peripheral blood mononuclear cell inflammation mediators ,PKC and NF-κB in MHD patients progressively .Methods1 Peripheral blood mononuclear cells,from 6 healthy volunteers and 6 MHD patients were obtained after centrifugalization,5×105PBMCs/well seeded in 96 wells cell culture plate were divided into 5 groups for cultivating 12 hours.before cultivating,LPS were added in every well,adjusting LPS final concentration as 100ng/ml,A1 group:normal PBMC,B1 group:patient PBMC,C1 group: patient PBMC+1.0mmol/L CaCL2,D1 group: patient PBMC+1.25mmol/L CaCL2,E1 Group: patient PBMC+1.5mmol/L CaCL2.2 Cell morphologic changes before and after culture were observed,CRP and IL-6 of culture supernatant were detected by ELISA;calcium ion concentration within cells were measured by Fluo-3 AM;3 PKC and NF-κB within cells were detected by western blot.Results1 Cell appearance and amount before and after 12 hours culture didn,t show marked change in every groups;2 Discrepancies of calcium ion concentration within cells after 12 hours culture between every groups were not significant;3 There were not obvious difference about levels of CRP and IL-6 between A 1and B1 groups after 12 hours culture;so were levels of PKC and NF-κB;4 Compared with E1 group;the levels of CRP and IL-6 in C1 group were higher after 12 hours culture ;so were gray levelsof PKC and NF-κB.Conclusions1 By stimulus of LPS, Calcium chloride of three kind concentrations didn't affect form and amount of PBMC of MHD patients after 12 hours culture;2 Discrepancies were not significant about effects of Calcium chloride of three kind concentrations on calcium ion concentration within PBMC of MHD patients after 12 hours culture under stimulus of LPS;3 Through stimulus of LPS,there were not obvious difference about levels of CRP and IL-6 from PBMC between healthy volunteers and MHD patients after 12 hours culture;so were levels of PKC and NF-κB;4 By stimulus of LPS,compared with 1.5mmol/L calcium chloride,1.0mmol/L calcium chloride significantly increased CRP and IL-6 production of PBMC,raised levels of PKC and NF-κB within PBMC in MHD patients after 12 hours culture.