The Level And Significance Of Serum Soluble E-selectin In Rats Model With Type 2 Diabetes Mellitus

Objective: To observe the density of Serum soluble E-sel- ectin(sE-selectin), HbA1C and fasting blood glucose(FBG) in a rat model with type 2 diabetes mellitus(T2DM) at different stage. We can know the level and the significance of sE-selectin, HbA1C and FBG and further explore the relationship between sE-selectin, HbA1C and FBG and the mechanism of development caused by E-selectin in the complication of T2DM.Then we can supply a new method for the therapy and prevention of T2DM's vascellum complication.Methods: 120 healthy 7 weeks old Wistar rats are included ,which belonged to inbred strain and weighed 180-220g,half are male and half are female.All the rats are fed adaptative for one week.Then all of them are randomly divided into two groups, named control group (CON) 50 rats, trial group (TRI)70 rats and both group of rats are half in female and male. The rats of CON are fed on standard diet,while the rats of TRI are fed on diet riched in fat and high energy. After four weeks, both two group rats have nothing by mouth for 8 hours.Then,the rats of TRI are injected intraperitoneally with a low-dose(30mg/kg) streptozoc- in (1%,solved to 0.1mmol/l sodium citrate buffer,pH4.4),while those of CON are injected intraperitoneally with sodium citrate buffer(0.1mmol/l, pH4.4). After one week, the test for bloting are taken. The rats whose FBG≥7.0 mmol/l and 2hBG≥11.1 mmol/l are successful to be the diabetic model,50 rats (half of each sex) are selected randomly to be as the diabetes mellitus group (DM).The fasting blood glucose and body weight(BW) are measured every two weeks. At week 0 (M0), week 4 (M4), week 8 (M8), week 12 (M12) and week 16 (M16), eight rats are picked up randomly and respectively from the two groups,HbA1c is measured(1μl,tail vein blood);then,the rats are anaesthesiaed by 10% chloral hydrate,the blood sample(5-7ml) are taken from the vena cava inferior and injected into normal test tube,waited for coagulation under room-temperature,centr ifugated by 3000/minute for 10 minutes.Then the serum from the blood are kept in -80℃refrigerator,at last sE-selectin are measured by ELISA at the same time.SPSS11.5 software pack is used for all the data to make statistical-analysis.Initially the homogeneity of variance among all the groups is analyzed.All the measurement data are denoted by mean±standard deviation.One-way analysis of variance (A- NOVA) and q test are used to establish significance,while analy- sis of line correlation is used to determine the relationships of variate. We took P<0.05 as statistic significant level.Results1 There are no animal dead during the whole experiment.The weight of rats in CON before the set-up of the diabetes model increase graduallyand FBG of which are normal (3.46±0.33mmol/l).The weight of TRI before the success of diabetes increase faster than which of CONand FBG of which are at the normal level(4.45±0.46mmol/l),too.2 The weight of rats of CON increase stably.There are no significant difference in sE-selectin,FPG and HbA1c in the rats of CON during the whole experimental course (P>0.05)(Table 1).3 The weight of rats in DM increase stably during M0-M8.After M8,the accrescence speed of weight became slower even the weight of rats are slower than that of before.There are no significant difference in statistics between CON and DM at the same time(Table 2,3;Fig.1).FPG of rats keep a higher level (BG>15mmol/l)(Table 3;Fig.2).4 HbA1c of rats in DM become higher gradually from 5.33±0.40%(M0) to 11.00±0.65%(M16).Furthermore,HbA1c is obvious higher than that of in previous experimental period at M8,M16(Table 4;Fig.3).5 The level of sE-selectin of rats in DM are higher than those of in CON at M0,M4,M8,M12,M16 and there are distinguished difference between CON and DM in statistic (p<0.05).With the development of rats in DM,the level of sE-selectin is higher and higher from 22.66±2.19ng/ml to 47.41±3.89ng/m l.Furthmore,the level of sE-selectin at M8,M16 in DM is higher than that of the previous time (p<0.05),while, there are no distinguished difference at M4,M12 compared to that of the level of sE-selectin at the previous time(Table 5;Fig.4).6 Both sE-selectin and HbA1c become higher and higher during the whole experiment period, and have positive correlation between sE-selectin and HbA1c in the rats of DM group (p<0.01)(Table 6;Fig.5).ConclusionsThe result of this research shows that the level of sE-select in a rat model with type 2 diabetes mellitus increases gradually as the accrescence of the age of rats.There was a strongly positive correlation between sE-selectin and HbA1C in the diabetic rats. The level of sE-selectin starts to advance as soon as HbA1c is higher than 5% and significantly advances as soon as HbA1c is higher than 8%, that is to say, the level of sE- selectin becomes higher gradually with the increase of HbA1c. These provide a theory evidence for the initial prevention and therapy of type 2 diabetes mellitus and its complication and the study of those pathogenesis.The high blood glucose leads to the accrescence of AGEs,while AGEs stimulates to generate a quantity of cytokine which induce the expression of E-select- in.The accrescence of E-selectin promotes that white blood cell (WBC) adhere to vaso-endothelial cell and take part in the occurrence and development in the great vessels complication of T2DM;at the same time,E-selectin activates WBC,then WBC releases oxygen-deri ved free radicals.So endothelial tissues are damaged and the blood platelet accumulates.These changes lead to micro-thrombus and vascular smooth muscle(VSM) are stimulated to hyperplasy.All these changes induce and promote the occurrence and development in the capillary vessel complication of T2DM.So,It is helpful to patients with T2DM in evaluating patient's condition and estimating prognosis to monitor the level of sE-selectin.Control of HbA1c as strict as possible and supplying the monoclonal antibody of anti-adhe- sion molecule (anti-sE-selectin medicine) as early as possible could decrease the expression of sE-selectin and the adhesion function of WBC indirectly.These could be helpful in preventing the occurrence and development of the vascellum complication in T2DM and increasing the level of therapy to 2DM.