当归补血汤干预化疗贫血小鼠的实验研究

【作者】 白媛媛；

【导师】 初杰；

【作者基本信息】 辽宁中医药大学 ， 方剂学， 2011， 硕士

【摘要】 目的:本实验研究借助环磷酰胺所致化疗贫血小鼠模型,考察当归补血汤对化疗性贫血小鼠的干预作用,比较当归补血汤高、中、低三个剂量水煎液干预化疗贫血小鼠的效应强度,为临床应用提供实验依据。材料与方法:按外周血血红蛋白浓度随机分组的原则,将60只BALB/c小鼠分为5组,分别是空白组、模型组、当归补血汤高、中、低剂量组。采用预防性给药方式,自实验前一天起,模型组给以蒸馏水,其他给药组给以相应药液灌胃,直到实验结束。实验第1天开始造模,造模组小鼠于实验第1、3天给模型组小鼠无菌腹腔注射环磷酰胺150mg/kg,第5天腹腔注射环磷酰胺200 mg/kg,每天注射1次,连续给药3次。第15天取材,眶静脉取抗凝血,采用全自动五分类多物种血液分析仪检测白细胞(WBC)、红细胞(RBC)计数及血红蛋白(Hb)浓度;摘眼球取血后,沉淀离心,提取血清,用ELISA法检测血清中的白细胞介素-2(IL-2)的浓度;脱颈处死小鼠后,立即解剖,取出肾脏,制成肾组织匀浆液,用ELISA法检测肾组织匀浆中促红细胞生成素(EPO)浓度;取小鼠一侧股骨,用6号针头以1ml生理盐水冲出骨髓细胞,4号针头过滤制成单个骨髓细胞悬液,用ELISA法检测骨髓中粒/巨噬细胞集落刺激因子(GM-CSF)浓度;取出脾脏,用电子天平称重,计算脾脏指数。结果:1模型组小鼠在第3次腹腔注射环磷酰胺后出现精神萎靡,行动迟缓,对外周刺激反应迟钝、蜷缩、弓背、毛蓬竖、少光泽,面、唇、眼、耳、尾部呈苍白色等现象;外周血象显示:与空白组比较,模型组小鼠白细胞、红细胞计数及血红蛋白浓度均显著下降,且血红蛋白浓度下降了正常值的34%。这表明了化疗贫血小鼠模型制备的成功。2当归补血汤各剂量组均能改善化疗贫血小鼠精神萎靡,行动迟缓,对外周刺激反应迟钝、蜷缩、弓背、毛蓬竖、少光泽,面、唇、眼、耳、尾部呈苍白色等现象,其中中剂量组改善程度最为明显。3与模型组相比,当归补血汤各剂量组均能提高外周血象白细胞、红细胞计数、血红蛋白浓度,其中当归补血汤中剂量水煎液的疗效最佳。4与模型组相比,当归补血汤各剂量组均能提高化疗贫血小鼠粒/巨噬细胞集落刺激因子(GM-CSF)、促红细胞生成素(EPO)浓度,促进造血祖细胞增殖,从而提高骨髓抑制小鼠的外周血象白细胞、红细胞计数及血红蛋白浓度,其中当归补血汤中剂量组疗效最佳。5与模型组相比,当归补血汤各剂量组均能提高化疗贫血小鼠血清中白细胞介素-2(IL-2)浓度和脾脏指数,从而提高化疗贫血小鼠的免疫功能,其中当归补血汤中剂量组疗效最佳。结论:1当归补血汤高、中、低三个剂量组对化疗药环磷酰胺所致的骨髓抑制和免疫抑制等毒副作用均有拮抗作用。2当归补血汤中剂量改善化疗骨髓抑制和免疫抑制优于当归补血汤的高剂量与低剂量。更多还原

【Abstract】 Purpose:This experiment chooses chemotherapy drugs cyclophosphamide to establish chemotherapy anemia model mice, observe different doses of Dangguibuxue decoction’s effect on chemotherapy anemia, compares the intervention of high, middle and low doses of Dangguibuxue decoction on chemotherapy anemia, and provide laboratory basis for clinical use.Material and method: 60 BALB/c mice were divided into five groups according to the peripheral blood cell count and hemoglobin randomly: the blank group, the model group, angelica blood soup high, medium and low dose group. before the experiment, preventively inject distilled water into mice’stomach in model group , other groups were injected different medicine until the end of experiment. making mould form experimental day 1.In experimental day 1&3 day to model mice sterile Irrigate stomach with cyclophosphamide 150mg/kg, In experimental day 5 adds cyclophosphamide to 200 mg/ kg, daily 1 times, continuous three days. experimental day 15 take materials, take anticoagulant from orbitofrontal vein, using automatic five classification many species blood analyzer WBC (WBC), detecting red blood cells (RBC) count and hemoglobin (Hb) concentration; After picking eyeball take blood sedimentation centrifuge, extraction, with ELISA test serum, serum interleukin - 2 (IL - 2) concentration; Take off immediately after death mice neck dissection, remove kidneys, make kidney tissues evenly grout,test erythropoietin (EPO) concentration by ELISA; Take thigh bone mice form the mice, filtrate marrow cells with 1ml NS by No. 6 injector, apart bone marrow cells and make levitation liquid, test concentration of marrow grain/ (GM-CSF) with ELISA; Remove the spleen, weighing with electronic balance and calculate the spleen index.Results:1. The mice in model group were insensitive to peripheral stimulation , spirits were drooping, feather were less sheen, curling up the body, arching the back, face, lips, eyes, ears, tail are pale etc. because of injected cyclophosphamide. Peripheral blood test shows: white, red blood cells in mice model group were drop than the mice in blank group. And the hemoglobin concentrations level dropped to 34% of the normal. This suggests that proof the chemotherapy anemia model mice preparation of success.2. The groups of different doses of dang gui bu xue tang improved immune function of chemotherapy anemia mice, The symptom just like insensitive to peripheral stimulation , spirits were drooping, feather were less sheen, curling up the body, arching the back, face, lips, eyes, ears, tail are pale were improved.3. Compared with model group, different doses of dang gui bu xue tang, the count of red and white blood cells, content of hemoglobin was enhanced. Group of medium dose attain the best effect.4 .Compared with model group, the different doses of dang gui bu xue tang are improvement chemotherapy anemia mice grain/macrophage stimulating factor (GM-CSF), erythropoietin (EPO) level, promote the hemopoietic progenitor cell to procreate, thus improving the level of marrow of peripheral blood on mice like red and white, the count of the hemoglobin content of blood, group of medium doses gain the best effect.5 . Compared with model group, each dose group of Dangguibuxue decoction improved IL– 2 level in serum and spleen index of chemotherapy anemia mice, thus improved the immune function of chemotherapy anemia mice. group of medium doses gain the best effect.Conclusion:1. The three doses of Dangguibuxue decoction can inhibit the side effects of bone marrow suppression and immunosuppression caused by cyclophosphamide.2 .For relieving bone marrow suppression and immunosuppression, the middle dose is the best among the three doses of Dangguibuxue decoction.更多还原