Marhoffer W, Stein M, Schleinkofer L, Federlin K
Clinic of Rheumatology Wiesbaden II, Germany.
J Biolumin Chemilumin. 1994 May-Jun;9(3):165-70. doi: 10.1002/bio.1170090310.
In this study neutrophil (PMN) phagocytic capacity was investigated using a conventional radiometric ingestion assay (IN) in comparison with PMN respiratory burst activity assessed by luminol-enhanced chemiluminescence (LCL) in response to phorbolesters and LCL induction during phagocytosis of opsonized Staphylococcus aureus (STLCL) in diabetes mellitus and healthy controls. PMN ingestion was measured with 3H-thymidine-labelled S. aureus in a kinetic radiometric assay. LCL and STLCL were assessed in a parallel detecting microtitre-plate luminometer (MTP-Reader). PMN of diabetic subjects showed a highly significant reduction of peak LCL in response to PMA as well as during phagocytosis of S. aureus (STLCL) compared to non-diabetic controls (p < 0.001 respectively). PMN ingestion in diabetic patients (51.8 +/- 4.6%) was significantly reduced compared to controls (78.3 +/- 6.2%) (p < 0.01). The in vitro data displayed impaired PMN oxidative burst activity at glucose concentrations > or = 13.8 mmol/L, whereas PMN IN was significantly reduced at glucose levels > or = 27.75 mmol/L. The control group showed a positive correlation of peak LCL response and IN (p < 0.05) but not of STCL and IN; in diabetic patients this was also true, but did not reach statistical significance. The data obtained in this study clearly demonstrated impaired PMN respiratory burst activity and markedly reduced phagocytic PMN functions in diabetic patients ex vivo and in vitro as measured by LCL and by ingestion of 3H-thymidine-labelled S. aureus suggesting inhibitory effects of elevated glucose concentrations on various PMN-functions, which might be of clinical importance concerning altered host defence.
在本研究中,采用传统的放射性摄取测定法(IN)研究中性粒细胞(PMN)的吞噬能力,并与通过鲁米诺增强化学发光法(LCL)评估的PMN呼吸爆发活性进行比较,该活性是针对糖尿病患者和健康对照中佛波酯刺激的反应以及吞噬调理金黄色葡萄球菌过程中的LCL诱导(STLCL)。在动力学放射性测定中,用3H-胸腺嘧啶标记的金黄色葡萄球菌测量PMN摄取。在平行检测的微量滴定板发光计(MTP-Reader)中评估LCL和STLCL。与非糖尿病对照相比,糖尿病患者的PMN对PMA刺激以及吞噬金黄色葡萄球菌(STLCL)过程中的LCL峰值显著降低(分别为p < 0.001)。糖尿病患者的PMN摄取(51.8 +/- 4.6%)与对照(78.3 +/- 6.2%)相比显著降低(p < 0.01)。体外数据显示,当葡萄糖浓度≥13.8 mmol/L时PMN氧化爆发活性受损,而当葡萄糖水平≥27.75 mmol/L时PMN IN显著降低。对照组中LCL峰值反应与IN呈正相关(p < 0.05),但STCL与IN无相关性;糖尿病患者也是如此,但未达到统计学显著性。本研究获得的数据清楚地表明,通过LCL以及3H-胸腺嘧啶标记的金黄色葡萄球菌摄取测量,糖尿病患者体内外的PMN呼吸爆发活性受损,吞噬性PMN功能明显降低,提示高血糖浓度对各种PMN功能有抑制作用,这可能在宿主防御改变方面具有临床重要性。