2型糖尿病患者血糖波动状态与血管内皮损伤、血小板活化及PKCβ1表达的相关性

Correlation Research on Blood Glucose Fluctuation in Type 2 Diabetes Mellitus Patients and Vascular Endothelial Injury/Platelet Activation/PKCβ1 Expression

目的探讨2型糖尿病患者血糖波动状态与血管内皮损伤、血小板活化及蛋白激酶Cβ1(protein kinase Cβ1,PKCβ1)表达的相关性。方法 38例2型糖尿病患者在未进行药物干预措施调整前测定三餐前、三餐后2 h及睡前21:00 7个时间段指端毛细血管血糖,计算平均血糖波动幅度(mean amplitude of plasma glucose excursions,MAGE);分别采用血小板聚集仪及流式细胞术检测外周血二磷酸腺苷(ADP)诱导的血小板聚集率(platelet aggregation rate,PAG)及血小板膜蛋白CD62p表达水平;采用离子交换高压液相法检测糖化血红蛋白(Hb A1c)水平;采用酶联免疫吸附法(ELISA)检测血清E-选择素(E-selectin)、血管性血友病因子(von Willebrand factor,v WF)和PKCβ1水平;同时检测患者肝、肾功能及血脂水平,测量血压并计算体重指数。以Hb A1c为评价长期血糖控制的监测指标,MAGE作为评价血糖波动性指标,采用Pearson相关分析和多元线性回归分析评价患者血管内皮损伤标志物(血清E-selectin、v WF水平)及血小板活化指标(PAG和血小板膜蛋白CD62p表达)分别与MAGE及冠心病血瘀证目标分子血清PKCβ1水平的相关性;并分析PKCβ1水平与MAGE和Hb A1c水平的相关性。结果入组患者年龄、体重指数、病程、病史等一般资料与血清E-selectin、v WF、PKCβ1水平、PAG、血小板膜蛋白CD62p表达及MAGE均无相关性(P>0.05);血管内皮损伤标志物(血清E-selectin、v WF)及血小板活化指标(PAG、血小板膜蛋白CD62p表达)与MAGE均呈显著相关(r值分别为0.468,0.609,0.451,0.674;P<0.01);PKCβ1与MAGE和Hb A1c、血管内皮损伤标志物(Eselectin和v WF)及血小板活化指标(PAG和CD62p表达水平)均呈显著相关(r值分别为0.643,0.705,0.394,0.665,0.441,0.577;P<0.01)。结论冠心病血瘀证关键调控分子PKCβ1与糖尿病患者血糖波动状态下的血管内皮损伤程度和血小板活化聚集水平密切相关。

Objective To observe the correlation between blood glucose fluctuation in type 2 diabetes mellitus（ T2DM） patients and vascular endothelial injury / platelet activation / protein kinase Cβ1（ PKCβ1）. Methods Capillary blood was collected from finger tips of 38 T2 DM patients at 7 time points,i. e.,before 3 meals,2 h after3 meals,21： 00 pm before sleep. The mean amplitude of plasma glucose excursions（ MAGE） was calculated. The peripheral blood platelet aggregation rate（ PAG） induced by adenosine diphosphate（ ADP） and platelet membraneprotein level of CD62 p were determined by platelet fluorescent aggregometer and flow cytometry respectively.Hb A1 c was measured by ion-exchange high-performance liquid chromatography. Serum levels of E-selectin,von Willebrand factor（ v WF）,and PKCβ1 were detected by ELISA. Meanwhile,liver and renal functions,blood lipids were also measured. Their blood pressure was measured and body mass index（ BMI） calculated. By taking Hb A1 c as a monitored index for assessing long-term glucose control,MAGE as an indicator for assessing glucose fluctuation,the correlations between serum markers for vascular endothelial injury（ levels of E-selectin and v WF） / platelet activation indices（ PAG and CD62 p expression） and PKCβ1 level / MAGE respectively were analyzed using Pearson correlation analysis and multivariant Logistic regression. The correlations between PKCβ1 level and MAGE / Hb A1 c were also analyzed. Results In simple correlation analysis,there were no significant correlations between age /BMI / course of disease / medical history / serum levels of E-selectin / v WF / PKCβ1 / PAG / CD62 p expression and MAGE（P〉0. 05）. There were significant correlations between vascular endothelial injury markers（ E-selectin and v WF） / platelet activation indicators（ PAG,CD62 p expression） and MAGE（r = 0. 468,0. 609,0. 451,0. 674; P〈0. 01）. There were significant correlations between PKCβ1 and glucose assessment indicators（ MAGE and Hb A1c） / vascular endothelial injury markers（ E-selectin and v WF）,platelet activation indicators（ PAG and CD62 p expression）（r = 0. 643,0. 705,0. 394,0. 665,0. 441,0. 577; P〈0. 01）. Conclusion PKCβ1,the key regulatory gene of coronary artery disease with blood stasis syndrome,was closely related with the degree of vascular endothelial injury and aggregation level of platelet activation in T2 DM patients with blood glucose fluctuation.