Muszyńska-Lachota Izabela
Zakładu Biochemii i Chemii Pomorskiej Akademii Medycznej w Szczecinie, al. Powstańców Wlkp. 72, 70-111 Szczecin.
Ann Acad Med Stetin. 2002;48:255-69.
This study presents the results of the research on transcapillary filtration of plasma proteins in patients with non-insulin dependent diabetes mellitus (NIDDM). The aims of the study were as follows: (a) Examination of transcapillary filtration of plasma proteins in patients with NIDDM; (b) Search for correlations between metabolic status of diabetes, severity of diabetic microangiopathy and penetration of individual protein fractions into the blister fluid; (c) Examination of the influence of nonenzymatic glycation on albumin fragmentation and its transcapillary filtration. It was also decided to check whether changes in the blister fluid would appear earlier than fundal fluorescein angiography or microalbuminuria, two markers of diabetic microangiopathy. The study enrolled 34 patients with type II diabetes mellitus, aged 35-78 years, with a mean disease duration of 12.5 years (SD = 7.8). The control group included 10 non-diabetic volunteers aged 31-76 years. The mean age in both groups did not differ significantly. The cantharidin blister method was used to study the permeability of capillary vessels. The following laboratory tests were done: concentrations of serum urea, creatinine, glucose, fructosamine, glycated hemoglobin, cholesterol, and triglycerides. Electrophoretic separation of proteins in plasma and blister fluid was performed and the concentration of fructosamine in fluid was measured. Routine urinalysis was supplemented with microalbuminuria. The eye fundus was inspected and fundal fluorescein angiography was done. The influence of nonenzymatic glycation on protein fragmentation and its possible impact on the permeability of capillary vessels was determined with in vitro glycation of human serum albumin. In patients without diabetic retinopathy the Cb:Cs ratio for beta-globulin turned out to be significantly lower than in patients with proliferative retinopathy. Patients with normoalbuminuria had the Cb:Cs ratio for alpha 2-globulin significantly lower than patients with urinary albumin concentrations > or = 165 mg/L. When the group was divided with regard to serum fructosamine concentrations, the subgroup with fructosamine concentrations lower than 3 mmol/L revealed a significantly higher Cb:Cs ratio for fructosamine than the subgroup with fructosamine > or = 3 mmol/L. Concentrations of total protein, albumin, beta-globulin and gamma-globulin in blister fluid correlated positively with glycemia in the control group. Blister fluid concentration of beta-globulin correlated positively with HbAIC concentration. In the study group, total protein content in blister fluid correlated negatively with duration of diabetes and level of glycosuria. Concentrations of blister fluid alpha 1-globulin and gamma-globulin correlated negatively with glycosuria. Concentrations of blister fluid beta-globulin correlated negatively with duration of diabetes. Electrophoretic separation of in vitro SDS glycosylated human serum revealed no additional fraction in the incubated samples. Electrophoretic separation of in vitro glycosylated albumin disclosed co-migration of albumin monomers incubated for varying times with glucose.
本研究展示了非胰岛素依赖型糖尿病(NIDDM)患者血浆蛋白经毛细血管滤过的研究结果。本研究的目的如下:(a)检查NIDDM患者血浆蛋白的经毛细血管滤过情况;(b)寻找糖尿病代谢状态、糖尿病微血管病变严重程度与各个蛋白组分渗入水疱液之间的相关性;(c)检查非酶糖基化对白蛋白片段化及其经毛细血管滤过的影响。还决定检查水疱液中的变化是否会比糖尿病微血管病变的两个标志物——眼底荧光血管造影或微量白蛋白尿出现得更早。本研究纳入了34例II型糖尿病患者,年龄在35 - 78岁之间,平均病程为12.5年(标准差 = 7.8)。对照组包括10名年龄在31 - 76岁之间的非糖尿病志愿者。两组的平均年龄无显著差异。采用斑蝥素水疱法研究毛细血管通透性。进行了以下实验室检查:血清尿素、肌酐、葡萄糖、果糖胺、糖化血红蛋白、胆固醇和甘油三酯的浓度。对血浆和水疱液中的蛋白质进行电泳分离,并测量液体中果糖胺的浓度。常规尿液分析补充微量白蛋白尿检测。检查眼底并进行眼底荧光血管造影。通过人血清白蛋白的体外糖基化确定非酶糖基化对蛋白质片段化的影响及其对毛细血管通透性的可能影响。在无糖尿病视网膜病变的患者中,β球蛋白的Cb:Cs比值显著低于增殖性视网膜病变患者。正常白蛋白尿患者的α2球蛋白Cb:Cs比值显著低于尿白蛋白浓度≥165 mg/L的患者。当根据血清果糖胺浓度对该组进行划分时,果糖胺浓度低于3 mmol/L的亚组的果糖胺Cb:Cs比值显著高于果糖胺≥3 mmol/L的亚组。对照组水疱液中总蛋白、白蛋白、β球蛋白和γ球蛋白的浓度与血糖呈正相关。水疱液中β球蛋白浓度与糖化血红蛋白A1C浓度呈正相关。在研究组中,水疱液中总蛋白含量与糖尿病病程和糖尿水平呈负相关。水疱液中α1球蛋白和γ球蛋白浓度与糖尿呈负相关。水疱液中β球蛋白浓度与糖尿病病程呈负相关。体外SDS糖基化人血清的电泳分离在孵育样品中未发现额外组分。体外糖基化白蛋白的电泳分离显示,与葡萄糖孵育不同时间的白蛋白单体发生了共迁移。