Serum Soluble L-selectin Level And Signifacance In Type 2 Diabetic Rats

Objective: At present, more and more people have gotten type 2 diabetes mellitus . The chronic complications of diabetes mellitus play an important role in the prognosis of diabetic patients .With the depth of the studies ,people paid more attention to the role of adhesion molecules .L-selectin were included in selectin family.Nowadays research have proved that L-selectin has important role in diabetes chronic complication,which through promoting leucocyte to adhere to platelet and endothelial cell, result in endodermis damage and form microthrombus .But there were little report up to now about L-selectin changes and an influence on factor of that in the diabetes condition .The trial will find out the changs of the leukoeyte adhesion molecules, observe the relationship among blood lipid , HbA1c and sL-selectin, discuss the role of selectin in the diabetic mechanism and diabetic mornitoring through duplicating type 2 diabetic rat models, detecting expression of diabetic rats selectin and determining blood lipid , HbA1c in the different disease course.Methods : 120 healthy 7 weeks old Wistar rats were included ,which weighed 180-220g ,and half were male and half were female(bought from the Experimental Animal Center of Hebei Medical University) . The rats were randomly divided into two groups : control group(CON) with 50 rats and trial(TIR) group with 70 rats after they were fed one week. The rats of control group were fed on standard diet ,which included protein 23%,carbohydrate 50%,fat 5% ,while the rats of trial group were fed on diet enriched in fat in order to induce insulin resistance ,which included protein 15%,carbohydrate 51%,fat 25% .After 4 weeks ,the rats of trial group were injected intraperitoneally with a low-dose streptozocin(25mg/kg) to induce hyperglycemia ,and at the same time ,the rats of control group were injected intraperitoneally with sodium citrate buffer once a week.Two weeks later ,the glucose tolerance was assayed to determine the set-up of diabetic model .Both of the two groups were fasted for 10-12 hours, measured the fast blood glucose(FBG) by the tail vein, and then the high carbohydrate glucose solution at the concentration of 50% according to 2g/kg were perfused into the stomach of all the rats .2 hours later ,the same method was used to measure the 2 hours blood glucose(2hBG) . The rats whose FBG≥7.0 mmol/l or 2hBG≥11.1 mmol/l were successful to be the diabetic model ,from which 50 rats (half of each sex) were selected to the diabetes group(DIA) ;10 rats were randomly rilled in CON and DIA at 0,4,8,12,16 week end .The rats of CON were fed on standard diet and the rats of DIA were fed on diet enriched in fat .The rats were weighed and the fasting blood glucose was measured every two weeks. sL-selectin ,blood lipid and HbA1c were measured regularly at 0,4,8,12 and 16 weeks end in DIA.After fasting for 10-12 hours ,the rats to be measured were anaesthesiaed ,then the blood sample were taken from the vena cava inferior ,5-7ml of the blood samples waited for coagulation under room-temperature ,then centrifugated by 3000/minute for 10 minutes ,obtained the serum from the blood ,the serum samples were kept in -80℃refrigerator ,then all samples be measured sL-selectin by ELISA at the same time, meanwhile, blood lipid and HbA1c will be detected.SAS6.10 software pack were used for all the data to make statistical-analysis .Initially the homogeneity of variance among all the groups was analyzed .All the measurement data was denoted by mean±standard deviation and students t test was used to establishsignificance . We took P<0.05 as statistic significant level .Result: 1Rats did not die in the whole trial. Before the diabetic model, the weight of rats of CON increased gradually ,and FBG,HbA1c of which were normal with disease development.The weight of DIA increased faster than which of CON, and have no statistical significance, FBG of which were mormal,too .2 After the set-up of the diabetes model , FBG of the rats of CON were stable and normal . With disease development, there were no significant differences among the different time spots, including sL-selectin, blood lipid and HbA1c (P>0.05) . FBG of the rats of DIA were high level ,and hadn't increased or decreased obviously (P>0.05) .There were no chage in two rats group at initial weight ,but weight regulally increased with time extend .From three week, weight of the rats of DIA increased faster than those of CON, the difference increased gradually until the trial end.3 SL-selectin of rats of DIA were higher than that of CON at 0 week.With DM development, the concentration of sL-selectin higher gradually. There were no obvious differences among 4,8,12,16week about sL-selectin .There were significant differences between 4,8,12 ,16 and 0 week.4 With disease grow, LDL-C of DIA lower gradually and HDL-C of DIA higher gradually than that of CON, their difference were statistical significance .5 LDL-C have positive correlation with sL-selectin, HDL -C have negative correlation with sL-selectin , and HbA1c have positive correlation with sL-selectin in DIA. Conclusion: The result of the trial suggest that sL-selectin were higher than that of CON. With DM development, the concentration of sL-selectin lower gradually. LDL-C, HbA1c have positive correlation with sL-selectin, HDL-C have negative correlation with sL-selectin through measured LDL-C,HDL-C and HbA1c of type 2 diabetic rats.The increase of expression of selectin that were a maker about leukoeyte active. The measure of sL-selectin can make as early maker of DM because leukoeyte adhesion molecules associated with the progression of DM. Meanwhile, DM have disorder about glucose and blood lipid ,2DM with high blood lipid happen after high glucose,LDL-C, HDL-C were correlation with HbA1c. HbA1c can make adcanced glycation end products(AGEs) increase in body ,high glucose autoxidation produced a great deal oxygen free radical ,stimulated leukoeyte and vascular endothelial cells to release some cytokine ,induced leukoeyte to express and release selectin. Reseach their role among them, which can expound mechanism of DM , provide beneficial thread for effective preventment DM.