温病气分证、营血分证大鼠模型血液流变学、D-二聚体及T细胞亚群的变化规律研究

Research on Changes of Blood Rheology,D-dimer and T cell subsets of Rat Models with Qi Stage Syndrome and Yingxue Stage Syndrome of Febrile Disease

目的观察大鼠气分证、营血分证血液流变学、D-二聚体及T细胞亚群的变化规律,探讨气分证、营血分证的病机实质,并为温病的治疗提供实验依据。方法将清洁级雄性SD大鼠30只随机分成三组:空白组、气分组、营血分组,采用尾静脉注射大肠杆菌内毒素的方法复制气分证及营血分证模型,应用粘度计测定全血粘度及血浆粘度,以压积管检测血沉及红细胞压积,采用酶联免疫吸附试验法(ELISA法)检测大鼠外周血D-二聚体水平,通过流式细胞仪测定大鼠外周血CD4+、CD8+阳性细胞数及CD4+/CD8+比值。结果与空白组相比,气分组及营血分组全血粘度、血沉显著升高(P<0.01),红细胞压积显著降低(P<0.01),而营血分组血浆粘度升高(P<0.05),营血分组与气分组比较,全血粘度、血沉显著升高(P<0.01),血浆粘度升高(P<0.05);与空白组相比,气分组与营血分组血浆D-二聚体水平显著上升(P<0.01),并且营血分组D-二聚体水平明显高于气分组(P<0.01);与空白组相比,气分组及营血分组CD4+阳性细胞数上升,而营血分组显著高于空白组(P<0.01),气分组及营血分组CD8+阳性细胞数显著下降(P<0.01),此外,气分组及营血分组与空白组相比,CD4+/CD8+比值明显升高(P<0.01),并且营血分组高于气分组(P<0.05)。结论血液流变学、D-二聚体及T细胞亚群的变化规律为温病气分证、营血分证的病机实质提供客观指标,并为临床提供诊疗依据。

Objective To observe changes of blood rheology,D-dimer and T cell subsets of Rat Models with Qifen Syndrome and Yingxuefen Syndrome of febrile disease,to investigate pathogenic essence of Qi Stage Syndrome and Yingxue Stage Syndrome and provide experimental evidence for the treatment of febrile diseases. Methods 30 SD rats were randomly divided into three groups：control group,Qi Stage Syndrome group and Yingxue Stage Syndrome group. The model rats were injected LPS from auricular vein to make the Qi Stage Syndrome and Yingxue Stage Syndrome. Viscometer was applied to test whole blood viscosity and plasma viscosity,hematocrit tubes were used to detect ESR and hematocrit,enzyme-linked immunosorbent assay（ ELISA） assay was used to test D-dimer levels in peripheral blood and flow cytometry was used to test CD4＋,CD8＋positive cells and CD4＋/ CD8＋ratio in peripheral blood of rats. Results Compared with the control group,the whole blood viscosity and erythrocyte sedimentation rate of Qi Stage Syndrome group and Yingxue Stage Syndrome group were significantly higher（P〈0. 01）,hematocrit decreased significantly（P〈0. 01）,and plasma viscosity of Yingxue Stage Syndrome group elevated（P〈0. 05）. Compared with Qi Stage Syndrome group,whole blood viscosity and ESR were significantly higher（P〈0. 01）,plasma viscosity increased（P〈0. 05）.Compared with the control group,D-dimer levels in the plasma of Qi Stage Syndrome group and Yingxue Stage Syndrome group increased significantly（P〈0. 01）,and D-dimer levels of Ying Stage Syndrome group was significantly higher than that of Qi Stage Syndrome group（P〈0. 01）. Compared with the control group,CD4＋positive cells of Qi Stage Syndrome group and Yingxue Stage Syndrome group increased,Yingxue Stage group was significantly higher than the control group（P〈0. 01）. CD8＋positive cells of Qi Stage Syndrome group and Yingxue Stage Syndrome group was significantly decreased（P〈0. 01）. In addition,compared with the control group,CD4＋/ CD8＋ratio of Qi Stage Syndrome group and Yingxue Stage Syndrome group were significantly higher（P〈0. 01）,and Yingxue Stage Syndrome group was higher than Qi Stage Syndrome（P〈0. 05）. Conclusion Changes of blood rheology,D-dimer and T cell subsets provide objective indicators for the pathogenic essence of Qi Stage Syndrome and Yingxue Stage Syndrome and basis for clinical diagnosis and treatment.