海藻糖负载对人红细胞膜的影响

Effect of Trehalose-loading on Red Blood Cell Membrane

本研究通过对海藻糖负载后红细胞膜各项理化指标检测,评价海藻糖对红细胞膜的保护作用。以海藻糖负载红细胞为实验组,未负载海藻糖红细胞为对照组,在不同渗透压的NaCl溶液中,检测2组红细胞膜渗透脆性变化,流式细胞术和红细胞变形仪分别检测2组红细胞膜的完整性和变形性。结果显示:对照组红细胞在渗透压为160 mOsm的NaCl溶液中溶血率为50%,而实验组红细胞出现50%溶血率的NaCl溶液渗透压为121.4 mOsm。流式细胞术检测结果显示,红细胞负载海藻糖后,细胞膜结合的AnnexinⅤ-FITC量很少,并且通过300 mOsm磷酸盐缓冲液的洗涤,破损细胞能被有效清除。负载后红细胞变形能力有所下降,2组之间存在显著差异(P<0.01)。结论:红细胞摄取海藻糖后能够在高渗环境中保持细胞膜稳定性和结构完整性。

This study was purposed to evaluate the effect of trehalose-loading on physiological and biochemistry properties of red blood cell （RBC） membrane. The samples were divided into the control group （RBC without trehalose loading） and the test group（ RBC with trehalose loading）. Osmotic fragility reaction was used to determine the osmotic fragility change of loaded RBC membrane in NaC1 solution of different osmotic concentration. Flow cytometry and deformeter were used to assay the integrality and deformability of the RBC, resectively. The results showed that the NaC1 solution osmotic concentrations were 160 mOsm and 121.4 mOsm, respectively when the haemolysis rate was 50% of the control group and the test group. Flow cytometry data demonstrated that incubation of RBC in a hypertortic trehalose solution resulted in a fraction of ceils with different complexity that attached to little Annexin V -FITC, and that it could be removed by washing and resuspending the RBC in an iso-osmotic （300 mOsm PBS ） medium. The deformability of the loaded RBC descend, the statistical difference was significant between control and test groups （ P 〈 0.01 ）. It is concluded that the membrane physiological and biochemistry stability and membrane integrality of RBC in a hyper osmotic pressure can be retained after trehalose loading.