| Objective: To measure the levels of serum soluble CD40 ligand(s CD40L) and P selectin in diabetic patients with lower extremity arterial disease(LEAD), simple diabetic patients and healthy people, as well as the levels after the diabetic patients with LEAD accepted the intervention, we explored the role of platelet activation in the development of diabetic with LEAD and its effect on restenosis after angioplasty or thrombosis to provide a clinical basis for the prevention of diabetic with LEAD and restenosis after angioplasty or thrombosis.Methods: Collection of specimens. We chose 25 cases of type 2 diabetes mellitus(T2DM) patients with LEAD(Fontaine degreeⅡb-Ⅳ) who were accepted the angioplasty or stent implantation and hosptitalized in endocrinology department of our hospital from June 2013 to December 2014 as PTA group. At the same time, another 25 cases of T2 DM patients(clinical and auxiliary examinations were used to exclude the diabetic complications and related diseases) in our hospital were chosen as T2 DM group. In addition, 25 cases of healthy people were chosen as Control group. The cases of PTA group and T2 DM group were in accord with the standard of WHO diabetes diagnosis in 1999.Exclusion criteria. All of the patients were excluded from hemorrhagic disease, bleeding or surgery in 2 weeks; cardiovascular and cerebrovascular diseases happened recently, infectious diseases, acute metabolic disorders, serious damage to the function of heart, kidney and hepar, tumor, blood diseases, autoimmune disease; the patients of Fontaine Ⅳin PTA group who were moist gangrene needed debridement.4 millilitre blood from cubital vein of all cases were taken after a 10-hour of fasting at 6:00am to test the levels of fasting blood glucose(FBG), glycosylated hemoglobin(Hb A1c), cholesterol(TC), triglyceride(TG), highdensity lipoprotein(HDL-C), low-density lipoprotein(LDL-C), s CD40 L and s P selectin. During the interventional treatment of PTA group, 4ml arterial blood(arterial blood before intervention) was taken after the artery sheath catheter placed, then the guide wire and catheter were passed through the stenosis or occlusion area of the vascular, 4ml arterial blood(Ischemic area prior-interventional) was taken at the distal of the lesion area. After balloon dilatation or stent implantation, another 4ml blood was taken at the distal of the lesion area(Ischemic area post-interventional). Patients of PTA group were supine to draw 4ml blood from cubital vein 24 hours and 7 days after the intervention(24h after intervention, 7d after intervention). The Blood samples collected were centrifuged at 1000r/min, 20 minutes conditions after standing at room temperature 2hours, the centrifugal radius was 15 cm. The supernatant was saved under-20 degree. To avoid repeated freezing and thawing to affect the test results, all the samples were subjected to the same manufacturers test kit. The age, sex, body mass index(BMI) of patients were recorded, and the ABI and Tc PO2 of PTA group were measured before and after the intervention, as well as T2 DM group. The serum levels of s CD40 L and s P selectin were measured by double antibody sandwich ELISA method, kits provided by Elabscience Biological Technology Co Ltd, in strict performed according to the instructions.SPSS13.0 software was used for statistical analysis. The measurement data was shown by means±standard deviation. The count data was using chi square test. Paired T-test was used for the comparision between before and after the intervention. The comparision among multiple groups was performed by analysis of variance, the comparision of one to another in the groups was performed by LSD test. Linear correlation analysis was used for the comparision among parameters. P < 0.05 was considered statistically significant.Results:1 The comparison of the general data in the three groups:The age, sex and BMI in three groups had no statistical significance(P>0.05).2 The levels of FBG and Hb A1 c among the three groups:In PTA group were respectively(8.68±0.83)mmol/L,(8.53±0.89)%; in T2 DM group were respectively(9.08±0.78)mmol/L,(8.69±0.86)%; in control group were respectively(5.22±0.52)mmol/L,(5.16±0.54)%. Compared with control group, the levels of FBG and Hb A1 c in T2 DM group and PTA group were significantly higher(T2DM group and control group: P<0.01; P<0.01; PTA group and control group: P<0.01; P<0.01). The levels of FBG and Hb A1 c in PTA group and T2 DM group had no statistical significance(P>0.05).3 The levels of TC, TG, HDL-C, LDL-C among three groups:In PTA group were respectively(5.46±0.81)mmol/L,(2.37±0.55)mmol/L,(1.06±0.21)mmol/L,(3.84±0.46)mmol/L; in T2 DM group respectively(5.02±0.62)mmol/L,(1.92±0.49)mmol/L,(1.20±0.19)mmol/L,(3.35±0.46) mmol/L; in control group were respectively(4.81±0.53)mmol/L,(1.52±0.38) mmol/L,(1.35± 0.21)mmol/L,(3.01±0.51)mmol/L. Compared with control group, the levels of TC, TG, LDL-C in PTA group were significantly higher(P<0.01; P<0.01; P<0.01), and the level of HDL-C was significantly lower(P<0.01); compared with control group, the levels of TG, LDL-C in T2 DM group were significantly higher(P<0.01;P<0.05), and the level of HDL-C was lower(P<0.05); compared with T2 DM group, the levels of TC, TG, LDL-C in PTA group were significantly higher(P<0.05; P<0.01; P<0.01), and the level of HDL-C was lower(P<0.05); All of the differences were statistically significant. But the levels of TC between T2 DM group and control group had no statistical significance(P>0.05).4 The serum levels of s CD40L:The s CD40 L levels of venous blood in PTA group, T2 DM group and control group were:(3.57±0.47)ng/ml,(3.09±0.62)ng/ml,(1.46±0.42)ng/ml, the level in PTA group was higher than that in T2 DM group and control group(P<0.01; P<0.01), and the level in T2 DM group was higher than that in control group(P<0.01). The levels of 24-hour and 7-day post-interventional venous blood were(5.58±0.58)ng/ml and(5.18±0.47)ng/ml, which were significantly higher than the level of pre-interventional venous blood(P<0.01; P<0.01), and the level of 7-day post-interventional venous blood was lower than that of 24-hour(P<0.01). The s CD40 L levels of pre-interventional arterial blood, ischemic area prior-interventional arterial blood and ischemic area post-interventional arterial blood in PTA group were(3.58±0.47)ng/ml,(3.93±0.44)ng/ml and(4.52±0.41)ng/ml, there was no significant difference about the level of pre-interventional between venous blood and arterial blood(P>0.05). The level of ischemic area prior-interventional arterial blood was higher than that of pre-interventional arterial blood(P<0.05), and the level of ischemic area post-interventional arterial blood was higher than that of ischemic area prior-interventional arterial blood(P<0.01). All of the differences were statistically significant.5 The serum levels of s P selectin:The s P selectin levels of venous blood in PTA group, T2 DM group and control group were:(45.07±7.94)ng/ml,(39.38±5.35)ng/ml and(29.34±4.52) ng/ml, the level in PTA group was higher than that in T2 DM group and control group(P<0.01; P<0.01), and the level in T2 DM group was higher than that in control group(P<0.01). The levels of 24-hour and 7-day postinterventional venous blood were(63.36±8.37)ng/ml and(50.84±7.38) ng/ml, which were significantly higher than the level of pre-interventional venous blood(P<0.01; P<0.01), and the level of 7-day post-interventional venous blood was lower than that of 24-hour(P<0.01). The s P selectin levels of pre-interventional arterial blood, ischemic area prior-interventional arterial blood and ischemic area post-interventional arterial blood in PTA group were(44.71±6.73)ng/ml,(50.17±7.63)ng/ml and(54.11±8.98)ng/ml, there was no significant difference about the level of pre-interventional between venous blood and arterial blood(P>0.05). The level of ischemic area priorinterventional arterial blood was higher than that of pre-interventional arterial blood(P<0.05), and the levels had no statistical significance between ischemic area post-interventional arterial blood and ischemic area prior-interventional arterial blood(P>0.05).6 The linear correlation analysis:The s P selectin level of pre-interventional venous blood was positively correlated with Hb A1 c. The s CD40 L level was positively correlated with Hb A1 c and TG. And both of s Pselectin and s CD40 L levels were negatively correlated with HDL-C(P<0.05). SP selectin and s CD40 L were positively related(r=0.717, P<0.01), the difference was statistically significant.7 The levels of ABI and Tc PO2:The levels of ABI and Tc PO2 in T2 DM group were(1.13±0.11) and(49.12±7.62)mm Hg, which were significantly higher than the levels in PTA group, which were(0.60±0.11) and(26.45±6.15)mm Hg(P<0.01; P<0.01). The levels of ABI and Tc PO2 in PTA group after intervention were(0.86±0.13) and(47.84±8.04)mm Hg, which were significantly higher than the levels before intervention(t=16.691, P<0.01; t=12.436, P<0.01), the difference was statistically significant.Conclusions:1 Simple diabetic patients have already platelet activation in vivo. Accompanied by the occurrence of LEAD, the platelet activation achieves a further development for diabetic patients.2 The blood glucose levels, which were not up to the standard, and lipid metabolism disorder effect the function of platelets in vivo, which promote the activity of vascular lesions.3 The levels of s CD40 L and s P selectin after intervention were higher than the levels before intervention in PTA group, and the levels of 7-day after intervention had not returned to the preoperative levels. Lower extremity interventions can activate the platelet activation and aggravate the inflammatory resonse. There is important significance to inhibition of platelet activation and anti-inflammatory therapy in the treatment of restenosis after angioplasty or thrombosis. |
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