猪链球菌2型对红细胞的黏附及溶血作用

发布时间：2018-11-17 11:18

【摘要】：为探讨猪链球菌2型(Streptococcus suis type 2,SS2)感染过程中对血液红细胞(RBC)损害及其机理,本研究采取无特定病原微生物感染的猪血液制备RBC悬液,根据预试验选用SS2∶RBC感染比(MOI)为100的SS2,分别体外感染RBC 0.5、1、1.5、2 h,光镜下观察BRC的形态变化;用G iem sa染色、CF D A-SE荧光标记检测SS2对红细胞的黏附和损伤,流式细胞仪分析不同作用时间的黏附率,real-time PCR检测SS2相关黏附因子溶菌酶释放蛋白(MRP)、荚膜多糖(CPS)、甘油醛-3-磷酸脱氢酶(GAPDH)、纤连蛋白和纤维蛋白结合蛋白(FBPS)及溶血因子溶血素(Sly)的转录水平。结果显示,SS2感染后第0.5~1小时,其主要黏附RBC,细胞呈现棘突样病变,少量细胞肿胀,出现轻微溶血,黏附率分别为18.7%和46.3%;感染后第1.5小时,SS2不仅可以黏附RBC,且多数细胞溶血,RBC开始发生破裂,黏附率可达到91.7%;感染后第2小时,细胞几乎全部破裂。MRP和CPS2J转录水平在感染后第0.5~1小时增高,并与黏附率呈正相关,感染后第2小时表达水平下降;Sly转录水平在感染后第0.5~2小时持续增加,与溶血现象呈正相关;GAPDH和FBPS在感染过程中变化不显著(P0.05)。结果表明,SS2感染早期可对RBC产生黏附和溶血作用,先为黏附作用,后为溶血作用,且作用过程较快;主要参与黏附因子为MRP和CPS2J,溶血因子为Sly;GAPDH和FBPS是否参与SS2对RBC的黏附与溶血仍需证实;本试验结果为研究SS2传播和致病机制提供了重要的理论依据。
[Abstract]:In order to investigate the (RBC) damage of erythrocytes caused by Streptococcus suis type 2 (SS2) infection and its mechanism, RBC suspension was prepared from porcine blood without specific pathogenic microorganisms. According to the pre-test, the SS2, with SS2:RBC infection ratio of 100 to (MOI) was infected with RBC 0.5A 1. 5U for 2 h in vitro respectively. The morphologic changes of BRC were observed under light microscope. The adhesion and injury of SS2 to erythrocytes were detected by G iem sa staining, CF D A-SE fluorescent labeling, the adhesion rate of erythrocytes was analyzed by flow cytometry, and the SS2 related adhesion factor lysozyme releasing protein (MRP), was detected by real-time PCR. The transcriptional levels of (GAPDH), fibronectin, fibrin binding protein (FBPS) and hemolysin (Sly) in capsule polysaccharide (CPS), glyceraldehyde 3 phosphate dehydrogenase. The results showed that at 0. 5 h after SS2 infection, the main adherent RBC, cells showed spinous process lesions, a small number of cells were swollen and slightly hemolytic, the adhesion rates were 18. 7% and 46. 3%, respectively. At 1. 5 hours after infection, SS2 not only adhered to RBC, but also hemolysis of most cells. RBC began to rupture, and the adhesion rate reached 91.7%. At the 2nd hour after infection, almost all of the cells ruptured. The transcription levels of MRP and CPS2J increased 1 hour after infection, and were positively correlated with the adhesion rate, and the expression level decreased at the second hour after infection. The transcription level of Sly increased continuously at 0.5h after infection, which was positively correlated with hemolysis, while GAPDH and FBPS did not change significantly during infection (P0.05). The results showed that SS2 infection could produce adhesion and hemolysis to RBC in the early stage, first adhesion, then hemolysis, and the course of action was faster, and the main factors involved in the adhesion were MRP and CPS2J, hemolytic factor Sly;. It is necessary to confirm whether GAPDH and FBPS are involved in the adhesion and hemolysis of SS2 to RBC, and the results provide an important theoretical basis for studying the transmission and pathogenesis of SS2.
【作者单位】：中国农业大学动物医学院;中国农业大学生物学院;
【基金】：国家自然科学基金项目(31260616)
【分类号】：S852.611

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