摘要
背景:研究已证实血小板活化参与了糖尿病血管病变的发生发展。血小板膜糖蛋白Ib/IX/V复合物是血小板膜上主要的糖蛋白分子之一,是血管性血友病因子和凝血酶的受体,在血小板活化中起重要作用。目的:观察2型糖尿病患者血小板膜糖蛋白Ib/IX/V复合物及其组分糖蛋白Ibα表达的变化。设计:病例-对照实验。单位:华中科技大学同济医学院附属协和医院中西医结合科。对象:根据1999年世界卫生组织制定的糖尿病诊断标准选择2005-12/2007-01本院门诊就诊的外周血小板计数>50×109L-1的2型糖尿病患者51例,男28例,女23例,纳入对象在检查前两周内未服用影响血小板功能的药物。根据病情控制情况将患者分为控制良好组(n=25)和控制不良组(n=26);根据是否合并血管病变分为合并血管病变组(n=27)和无血管病变组(n=24)。选择同期本院健康体检者23名为健康对照组,年龄、性别与病例组匹配。纳入对象或家属对检测项目和试验知情同意。试验经医院伦理委员会批准。方法:患者均在清晨就诊时抽取空腹肘静脉血进行检测。健康体检者于来院体检时进行相同检测。①生化指标检测:由本院检验科完成空腹血糖、糖化血红蛋白和空腹面胰岛素的检测。②糖蛋白Ib/IX/V复合物及其组分糖蛋白Ibα的表达的检测:采集清晨空腹静脉血3mL,38g/L枸橼酸钠抗凝后10g/L多聚甲醛固定45min。取50μL固定后全血加至流式检测管,同时分别向不同管中加入20μL糖蛋白Ib/IX/V复合物单克隆抗体CD42a-b-c-d和PE标记的糖蛋白Ibα单克隆抗体CD42b,混匀后室温避光孵育30min,然后CD42a-b-c-d标记的样品中加入20μLFITC标记的羊抗鼠IgG,混匀后室温避光孵育30min。于FACS420型流式细胞仪上检测样本平均荧光强度。③血小板聚集率测定:参照文献检测血小板聚集率。主要观察指标:①生化指标。②糖蛋白Ib/IX/V复合物及糖蛋白Ibα的表达情况。③血小板最大聚集率。结果:实验纳入的51例2型糖尿病患者和23名健康体检者全部进入结果分析。①生化指标检测结果:控制良好组和控制不良组患者空腹胰岛素与健康对照组比较,差异有显著性意义(P<0.01)。控制不良组患者空腹血糖和糖化血红蛋白均明显高于健康对照组和控制良好组,差异有显著性意义(P<0.01)。②糖蛋白Ⅰb/Ⅸ/Ⅴ复合物和糖蛋白Ⅰbα表达情况的比较:控制良好组和控制不良组患者糖蛋白Ⅰb/Ⅸ/Ⅴ复合物和糖蛋白Ⅰbα的表达明显低于健康对照组,控制不良组明显低于控制良好组,差异均有显著性意义(P<0.05～0.01)。无血管病变组和合并血管病变组患者均明显低于健康对照组,差异有显著性意义(P<0.01)。无血管病变组和合并血管病变组患者糖蛋白Ⅰbα明显低于健康对照组,合并血管病变组明显低于无血管病变组,差异均有显著性意义(P<0.05)。糖蛋白Ⅰb/Ⅸ/Ⅴ复合物平均荧光强度与空腹血糖、糖化血红蛋白和空腹胰岛素呈明显负相关(r=-0.634,-0.573,-0.649,P<0.05);糖蛋白Ⅰbα平均荧光强度与空腹血糖和糖化血红蛋白呈明显负相关(r=-0.602,-0.543,P<0.05)。③血小板聚集功能的比较:2型糖尿病患者血小板最大聚集率较正常人明显升高,差异有显著性意义(t=-3.852,P<0.01),控制不良组明显高于控制良好组,合并血管病变组高于无血管病变组,差异均有显著性意义(P<0.05)。结论:2型糖尿病患者早期无血管病变时即存在血小板活化,发生血管病变后活化更明显,且血小板活化与血糖呈正相关。血小板糖蛋白Ⅰb/Ⅸ/Ⅴ复合物作为凝血酶和血管性血友病因子的受体,可能参与了2型糖尿病血管病变的发生发展。
BACKGROUND： It has been proved that platelet activation is involved in the development of diabetic angiopathy. Glycoprotein （GP） Ib/IX/V complex is one of the main platelet membrane glycoproteins, and the receptor of both von Willebrand Factor and thrombin. It plays a key role in the process of platelet activation.
OBJECTIVE： To observe the expression changes of GP Ib/IX/V complex and its component GP Ib a in patients with type 2 diabetes mellitus.
DESIGN： Case-control study.
SETTING： Department of Integrated Chinese and Western Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.
PARTICIPANTS： A total of 51 type 2 diabetic outpatients who visited Union Hospital were enrolled from December 2005 to January 2007. The diagnosis was based on the independent criteria from WHO in 1999. Of all the 51 patients, there were 23 females and 28 males, with a peripheral platelet count of over 50 × 10^9 L^-1. All the subjects had no history of administrating drugs two weeks before the examination, which would potentially influence platelet count. According to disease controlling condition, the patients were assigned to well controlled patient （WCP） group （n = 25） and poorly controlled patient （PCP） group （n =26）; and according to whether angiopathy was accompanied, diabetic patients were divided into vascular disease （VD） group （n =27） and non-vascular disease （NVD） group （n =24）. Meanwhile 23 healthy subjects were enrolled as normal control group. Informed consents were obtained from subjects and their relatives. The experiments were approved by the ethical committee of Union Hospital.
METHODS： Fasting venous blood was harvested from all the subjects＇ elbow on the early morning of visit day.① Biochemical analysis： Fasting plasma glucose （FPG）, glycosylated hemoglobin （HbAlc） and fasting plasma insulin （FINS） was measured by the Clinical Chemistry Department in Union Hospital.② Measurement of platelet membrane GP Ib/IX/V complex and its component GP Ib a ： First, 3 mL cubital fasting blood was drawn from each subject and was anti-coagulated with 38 g/L natrium citricum. After that, all samples were fixed with 10 g/L paraform for 45 minutes. Then 50 μL well-fixed blood was added into the polystyrene tube, meanwhile 20 μL monoclonal antibody, such as CD42a-b-c-d and PE-labeled CD42b, was respectively mixed gently with the blood sample and incubated at room temperature in dark for 30 minutes. Next, 20 μL FITC-labeled rat IgG was mixed with the sample containing CD42a-b-c-d and incubated equally. In the end all blood samples were analyzed by FACS420 flow cytometry and the results were expressed as mean fluorescence intensity （MFI）. ③ Platelet maximun aggregation rate （MAR） was detected according to reference.
MAIN OUTCOME MEASURES： ①Biochemical indicators;②The expressions of GP Ib/IX/V complex and GP Ib α ;③ Platelet MAR.
RESULTS： Fifty-one patients with type 2 diabetes mellitus and twenty-three healthy subjects were all involved in the result analysis.①There were significant differences in FINS in WCP group and PCP group compared with normal controls （P 〈 0.01）. FPG and HbAlc were significantly higher in PCP group compared with normal control group and WCP group （P 〈 0.01）.②Expressions ofGP Ib/IX/V complex and GP Ib a were significantly lower in WCP group and PCP group compared with normal control group （P 〈 0.01）, significantly lower in PCP group than in WCP group （P 〈 0.05）, and also significantly lower in VD group and NVD group compared with normal control group （P 〈 0.01 ）. Moreover the expression of GP Ib a in VD group and NVD group was significantly lower than that of normal control group （P 〈 0.05）, and it also significantly decreased in VD group compared with NVD group （P 〈 0.05）. MFI of GP Ib/IX/V complex had an obvious negative correlation with FBG, HbAlc and FINS （r =0.634, -0.573, -0.649, P 〈 0.05）, and GP Ib a MFI was obviously negatively correlated with FBG and HbAlc （r=0.602, -0.543, P 〈 0.05）.③Platelet MAR of diabetic patients were remarkably higher than in healthy subjects （t=3.852, P 〈 0.01）. Platelet MAR in PCP and VD groups were respectively higher than those in WCP and NVD groups （P 〈 0.05）.
CONCLUSION： Platelet activation exists in the early stage of type 2 diabetes mellitus without diabetic angiopathy, and is more obvious after diabetic angiopathy. There is a positive correlation between platelet activation and blood glucose. As a receptor of thrombin and von Willebrand Factor, GP Ib/IX/V complex may be involved in the development of diabetic angiopathy.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第7期1393-1396,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
