苜蓿皂苷对小白鼠体内溶血的研究

为了探究摄入苜蓿皂苷(alfalfa saponin, AS)是否会引起血管内溶血,选取180只健康小白鼠,按体重和性别随机分为6个处理,每个处理3个重复,每个重复10只。将1mL 0.0125、0.025、0.05、0.1、0.2 g·mL^-1皂苷分别灌胃Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ组小鼠,对照组灌胃等量的生理盐水(1 mL),试验期为30 d。检测红细胞数目(red blood cell, RBC)、形态、压积(hematocrit, HCT)、平均红细胞体积(erythrocyte mean corpuscular volume, MCV)、平均血红蛋白浓度(mean corpuscular hemoglobin, MCH)、血浆游离血红蛋白(free hemoglobin, FHb)、网织红细胞计数(reticulocyte count, RET)及红细胞渗透脆性最大、最小抵抗值(maximum/minimum osmotic fragility of red blood cells, OF_max/OF_min),采血和骨髓制作观察血涂片及骨髓涂片,制作实质性器官心、肝的切片。结果表明: 1)灌胃苜蓿皂苷后,小白鼠血常规中红细胞数目和血细胞比容均降低且Ⅴ组显著低于对照组(P<0.05),平均血红蛋白浓度均高于对照组且Ⅳ、Ⅴ组与对照组相比差异显著(P<0.05)。2)Ⅳ、Ⅴ组血浆游离血红蛋白含量极显著高于对照组(P<0.01);Ⅳ组网织红细胞计数极显著高于对照组(P<0.01)。3)随着灌胃剂量的加大,红细胞逐渐出现轻度大小不等,着色较深,形态异常的现象。4)红细胞渗透脆性最大抵抗值显著升高且差异显著(P<0.05),最小抵抗值有逐渐升高的趋势。5)灌胃后粒系比有不同程度的降低,Ⅴ组与对照组相比差异显著(P<0.05);红系比则有不同程度的升高,Ⅴ组与对照组相比差异显著(P<0.05);粒红比不但降低而且变化明显,Ⅳ、Ⅴ组显著低于对照组(P<0.05)。 6)灌胃对实质性器官没有影响但是组织更易碎。综上,高剂量的苜蓿皂苷能够引起血管内溶血。

Effects of alfalfa saponins on the blood and hematopoietic system of mice

In order to explore whether the intake of alfalfa saponins can cause intravascular hemolysis. A total of 180 mice were selected and divided into 6 treatment groups according to their body weight and sex, with 3 replicates in each treatment, and with 10 mice in each replicate. A 1 mL gavage containing 0.0125, 0.025, 0.05, 0.1, and 0.2 g·mL^-1 saponin, respectively was given to the mice of Groups Ⅰ, Ⅱ, Ⅲ, Ⅳ, and Ⅴ, and the remaining group (Control) was given 1 mL normal saline. The test period was 30 days. On the final day of the experiment, red blood cell (RBC), hematocrit (HCT), erythrocyte mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), free hemoglobin (FHb), reticulocyte count (RET), maximum osmotic fragility of red blood cells (OF_max) and minimum osmotic fragility of red blood cells (OF_min) were measured. Blood and bone marrow were collected to observe blood smear and bone marrow smear while heart and liver slices were made. The results show: 1) After intragastric administration of alfalfa saponins, RBC and HCT were decreased with values for Group Ⅴ being significantly lower than those of the control group (P<0.05). MCV was elevated compared with the control group, with Groups Ⅳ and Ⅴ being significantly different from the control group (P<0.05). 2) FHb values in Groups Ⅳ and Ⅴ and RET in Group Ⅳ were significantly higher than those in the control group (P<0.01). 3) With increase in the intragastric dose, the red blood cell appearance gradually changed, becoming slightly different in size, darker in color, and abnormal in shape. 4) OF_max was significantly increased (P<0.05) and OF_min sshowed a trend towards increase. 5) The proportion of granulocytes was reduced to varying degrees after gavage, and the erythrocyte proportion was increased. Thus, Granulocyte: erythrocyte ratio decreased markedly and in Groups Ⅳ and Ⅴ became significantly lower than in the control group (P<0.05). Compared with the control group, both OF_max and OF_min in Group Ⅴ differed significantly from the Control mice (P<0.05). 6) Parenchymal organs were not effected after gavage but were easily broken. In summary, a high dose of alfalfa saponins can cause intravascular hemolysis.