松果菊苷对细胞外组蛋白介导的细胞损伤及红细胞和血小板聚集的保护作用

Protective Effects of Echinacea on Extracellular Histone Mediated Cell Damage and Aggregation of Erythrocytes and Platelets

目的:探讨松果菊苷(ECH)对细胞外组蛋白介导的细胞损伤以及红细胞和血小板聚集的保护作用。方法:采用Calcein-AM和PI双染色法考察ECH与组蛋白共同使用时对组蛋白介导人微血管内皮细胞(HMEC)毒性作用影响以及ECH介导的HMEC损伤的逆转作用;考察ECH对组蛋白诱导的小鼠红细胞聚集作用的影响;进一步考察了ECH对组蛋白介导小鼠体内生物标志物(ALT、LDH、Crea)以及血液学参数(白细胞、血小板、红细胞、血红蛋白)变化的影响。结果:ECH对组蛋白介导的HMEC损伤作用呈剂量依赖性保护作用,并且ECH(100μg/mL)处理10min后可部分逆转组蛋白介导的HMEC细胞毒性作用;组蛋白可以加速未经处理的红细胞的自然聚集率,而加入200μg/mL的ECH溶液对红细胞聚集具有明显的抑制作用;ECH可以剂量依赖性的抑制血清中生物标记物循环丙氨酸转氨酶(ALT)、乳酸脱氢酶(LDH)和肌酐(Crea)表达,并显著增加小鼠体内的循环血小板、白细胞、红细胞和血红蛋白数量。结论:ECH可减轻组蛋白介导的细胞毒性,并抑制组蛋白介导的红细胞凝集以及血液学参数变化,ECH体外实验加上相关的体内模型,证实其可通过与细胞外组蛋白作用从而降低脓毒症患者器官损伤和死亡率的治疗潜力。

Objective:To observe the protective effects of Echinacea(ECH)on extracellular histone mediated cell damage and aggregation of erythrocytes and platelets.Methods:Calcein AM and PI double staining method was used to investigate the toxic effect of ECH on human microvascular endothelial cell(HMEC)and the reversal of HMEC injury induced by ECH.The effect of ECH on the aggregation of erythrocytes induced by histone,and the influence of mice internal biomarkers(ALT、LDH、Crea)and hematological parameters(WBC、Platelets、RBC and hemoglobin).Results:ECH showed a dose-dependent protective effect on HMEC injury by histone and could partially reverse the histone mediated cytotoxicity of HMEC.Histone could accelerate the natural aggregation rate of untreated red blood cells,while ECH(200μg/ml)could significantly inhibit the aggregation of red blood cells.ECH could be dose-dependent inhibited the expression of ALT,LDH and Crea,and increased the number of circulating platelets,leukocytes,erythrocytes and hemoglobin in mice.Conclusion:ECH can reduce histone mediated cytotoxicity,and inhibit histone mediated red blood cell agglutination and hematological parameter changes.Vitro experiment and related in-vivo model confirmed that ECH can reduce organ damage and mortality of sepsis patients through its effect on extracellular histone.