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目的探讨纳米羟基磷灰石(n-HAP)溶胶饱和溶液在淋巴细胞性白血病荷瘤小鼠(P388)体内抗瘤作用及其机制。方法 (1)n-HAP 溶胶饱和溶液的制备:高纯0.1 mol 氢氧化钙混悬液和0.06 mol 磷酸溶液,经中和反应制备出稳定的 n-HAP 溶胶饱和溶液,浓度为167 mg/L(室温、pH>7)。经透射电镜、X-衍射图谱和红外光谱测定其表征。(2)建立 P388实体瘤型荷瘤小鼠模型:将60只 P388荷瘤小鼠随机分为治疗组36只和对照组24只。治疗组包括:①n-HAP 低剂量组(35 mg/kg)、②n-HAP 高剂量组(53 mg/kg)、③联合用药组(n-HAP 53 mg/kg 与环磷酰胺35 mg/kg 联合应用),每组各12只。对照组分为阴性对照组(生理盐水0.1 ml/g)和阳性对照组(环磷酰胺35 mg/kg),腹腔注射,连续3 d;用细胞形态学、病理学和透射电镜等方法,观察药物对各组荷瘤小鼠生长、瘤体大小及细胞凋亡等影响。结果 (1)自制 n-HAP 溶胶饱和溶液,经透射电镜观测 n-HAP 形状为球形,平均粒径<50 nm。X-衍射图谱表明 n-HAP 为一种透明或液态的典型的非结晶结构,通过谱线增宽分析,粒径<20 nm(Scherrer 理论);X-衍射图谱还可见到 n-HAP 在800℃结晶作用下形成的磷灰石晶体结构。n-HAP 经红外光谱检测含氢氧化物离子结构,证实是羟基磷灰石纳米晶体。(2)动物体内实验:n-HAP 低剂量组、n-HAP 高剂量组和联合用药组对 P388实体瘤的抑制率分别为14.95%、32.67%和60.45%,与阴性对照组比较,差异均有统计学意义(P<0.01)。体外透射电镜结果显示,n-HAP 对 P388细胞有抑制生长、诱导凋亡作用。结论 n-HAP 溶胶饱和溶液对淋巴细胞白血病实体瘤荷瘤小鼠有显著的抑瘤作用,且呈明显的量效关系,联合用药抑瘤作用更为显著。n-HAP 有可能用于恶性血液病及肿瘤的治疗和联合化疗。
Objective To investigate the anti-tumor effect and mechanism of nano-hydroxyapatite (n-HAP) sol saturated solution in lymphocytic leukemia bearing mice (P388) in vivo. Methods (1) Preparation of n-HAP sol saturated solution: Highly pure 0.1 mol calcium hydroxide suspension and 0.06 mol phosphoric acid solution were used to prepare a stable saturated solution of n-HAP sol by neutralization at a concentration of 167 mg / L (Room temperature, pH> 7). The characterization was carried out by transmission electron microscopy, X-ray diffraction and infrared spectroscopy. (2) To establish P388 solid tumor-bearing mice model: 60 P388 tumor-bearing mice were randomly divided into treatment group 36 and control group 24. The treatment group included: ①n-HAP low-dose group (35 mg / kg); ②n-HAP high-dose group (53 mg / kg); ③the combination of n-HAP 53 mg / kg and cyclophosphamide 35 mg / Combined application), each group of 12. The control group consisted of a negative control group (saline 0.1 ml / g) and a positive control group (cyclophosphamide 35 mg / kg) for 3 days. The morphological, pathological and transmission electron microscope The effects of drugs on the growth, tumor size and apoptosis of tumor-bearing mice in each group. Results (1) Self-made n-HAP sol saturated solution, observed by transmission electron microscopy n-HAP spherical shape, the average particle size <50 nm. X-ray diffraction showed that n-HAP was a typical amorphous or liquid amorphous structure with a particle size of <20 nm (Scherrer theory) by X-ray broadening analysis; X-ray diffraction showed that n-HAP was at 800 Crystallization of apatite crystals formed under the crystal structure. n-HAP was detected by infrared spectroscopy with hydroxide ion structure, confirmed to be hydroxyapatite nanocrystals. (2) Animal experiments: The inhibitory rates of low dose n-HAP group, n-HAP high dose group and combination group on P388 solid tumor were 14.95%, 32.67% and 60.45%, respectively. Compared with the negative control group, There was statistical significance (P <0.01). In vitro transmission electron microscopy showed that n-HAP inhibited the growth of P388 cells and induced apoptosis. Conclusion The n-HAP sol saturated solution has significant anti-tumor effect on solid tumor-bearing mice with lymphocytic leukemia, and has a significant dose-effect relationship. The combined anti-tumor effect is more significant. n-HAP may be used for the treatment of haematological malignancies and tumors and in combination with chemotherapy.