2型糖尿病患者中性粒细胞趋化功能受损及可能机制

Neutrophil chemotaxis dysfunction and potential mechanisms in diabetes mellitus

目的:探讨2型糖尿病患者中性粒细胞趋化功能是否受损及其潜在机制。方法:按WHO诊断标准,招募确诊的2型糖尿病患者及健康同龄志愿者各45例,受检测的中性粒细胞均取自受试者肘静脉外周血。使用改良的琼脂糖下趋化模型检测两组患者中性粒细胞趋化功能,流式细胞术及免疫荧光法检测中性粒细胞膜表面P2X1受体表达水平。在干预实验中,分别使用稀释的2型糖尿病患者或健康对照的血浆(90%、20%),不同葡萄糖含量(5、12和25 mmol/L)的培养基,糖基化终产物(0.5、1、2和4 mmol/L)以及棕榈酸(50、100和200μmol/L)刺激健康对照中性粒细胞后检测趋化功能。结果:与健康对照组相比,2型糖尿病患者中性粒细胞趋化功能显著降低(P<0.001),膜表面P2X1受体表达明显增加(P<0.001),二者呈负相关(r=-0.60,P<0.001)。相较于对照组,高浓度2型糖尿病患者血浆显著抑制中性粒细胞趋化功能(P<0.001);不同浓度的糖基化终产物及葡萄糖对中性粒细胞趋化功能没有明显抑制(P>0.05);高浓度棕榈酸(200μmol/L)明显抑制中性粒细胞趋化功能(P<0.05),同时促进细胞膜表面P2X1受体表达(P<0.05),二者呈负相关(r=-0.67,P<0.05)。结论:2型糖尿病患者中性粒细胞趋化功能受损,高浓度棕榈酸通过P2X1受体抑制中性粒细胞趋化功能。

Objective:To investigate whether the chemotactic function of neutrophils in type 2 diabetes is impaired and its underlying mechanism.Methods:According to the WHO diagnostic criteria,a total of 45 patients with diagnosed type 2 diabetes and 45 healthy controls of the same age were recruited.The tested neutrophils were all collected from the peripheral blood of the subjects′cubital veins.The chemotactic function of neutrophils in the two groups was detected by the modified chemotaxis method under agarose,and the expression level of P2X1 receptor on the surface of neutrophil membrane was detected by flow cytometry and immunofluorescence.In the intervention experiments,the normal neutrophils were stimulated by diluted plasma of patients with type 2 diabetes or healthy controls(90%,20%),media with different glucose contents(5,12 and 25 mmol/L),end products of glycation(0.5,1,2 and 4 mmol/L)and palmitic acid(50,100 and 200μmol/L)and the chemotaxis were detected.Results:Compared with healthy controls,the chemotactic function of neutrophils type 2 diabetes was significantly reduced(P<0.001),and the expression of P2X1 receptors increased(P<0.001).There was a significantly negative correlation between them(r=-0.60,P<0.001).Compared with the control group,the patients′high-concentration plasma significantly inhibited the chemotactic function of neutrophils(P<0.001).Each concentration of glycation end products and glucose was not significantly inhibited neutrophils chemotaxis function(P>0.05).While high concentrations of palmitic acid(200μmol/L)significantly inhibited the chemotaxis of neutrophils(P<0.05),the expression of P2X1 receptors on the cell surface increased(P<0.05),and there was a significantly negative correlation(r=-0.67,P<0.05)between them.Conclusion:The chemotaxis of neutrophils is impaired in type 2 diabetes,and high concentrations of palmitic acid impairs the chemotaxis of neutrophils through P2X1 receptors.