摘要
目的纳米颗粒因为较小尺寸、较大的表面积使其具有特殊的活性和内在毒性。文中研究纳米ZnO对雄性小鼠的体内作用,观察其对雄性小鼠生殖系统的影响。方法45只6~7周龄的雄性ICR小鼠随机均分3组,每组15只,纳米ZnO处理组隔天腹腔分别注射200、500mg/kg的纳米ZnO,对照组注射等体积等渗盐水,共5次。停药1周后,测量心脏、肝、肾、脾、睾丸和附睾的器官系数,血清生化指标、睾酮和雌二醇水平;显微镜下观察附睾精子数量、活率、畸形率和睾丸内精子计数;主要器官做病理切片和HE染色;电子显微镜观察肝和睾丸内纳米颗粒的分布。TUNEL法检测睾丸生殖细胞凋亡。结果对照组和纳米ZnO200mg/kg组小鼠无死亡,500mg/kg组小鼠死亡2只。与对照组相比,纳米ZnO200mg/kg组器官质量系数均无明显改变（P〉0.05）;500mg/kg组小鼠脾和肾质量系数明显升高（P〈0.05）。纳米ZnO200mg/kg组丙氨酸氨基转移酶（ALT）、总胆红素（TBIL）、胆碱（CHO）、尿酸（UA）、肌酸激酶（CK）显著性改变,P〈0.05;500mg/kg组肝功能酶ALT、天门冬氨酸氨基转移酶（AST）、碱性磷酸酶（ALP）、TG、CHO,肾功能的指标尿素氮（BUN）、肌酐（Cr）、UA和心功能指标CK均明显升高,P〈0.05。与对照组比较,纳米ZnO处理组血清睾酮（T）和雌二醇（E2）水平未见显著变化（P〉0.05）。纳米ZnO200mg/kg组附睾精子计数和睾丸内精子计数均有所降低,但P〉0.05;活精子率降低、精子畸形率升高,P〈0.05;500mg/kg组上述指标变化差异均有显著性意义（P〈0.05）。TURNEL检测显示,纳米ZnO处理后睾丸生精细胞凋亡率增加（P〈0.05）。纳米ZnO200mg/kg组各器官病理切片未见明显改变;500mg/kg组病理切片可以看到肝细胞轻度肿胀,空泡化,见点状坏死。肾小管上皮细胞肿胀,核空泡变,核仁明显。脾红髓内多核巨细胞浸润。心、睾丸和附睾的病理切片HE染色观察未见明显改变。电镜可见纳米ZnO颗粒在肝细胞和睾丸细胞分布,并在溶酶体、线粒体等细胞器中聚集。结论腹腔注射纳米ZnO200mg/kg对小鼠的肝、肾和心功能有轻度影响,并影响活精子率和精子畸形率;500mg/kg纳米ZnO对小鼠的肝、肾和心功能有明显影响,影响小鼠精子数量和质量,诱导睾丸生精细胞的凋亡。
Objective While research on the potential toxic properties of nanomaterials is now increasing, the in vivo effects of zinc oxide （ZnO） on the main organs were emergent to know. In this study, the effects of nanometer ZnO on male mice, particularly the reproductive system were evaluated. Methods Forty-five male ICR mice, aged 6-7weeks, were randomly divided into control group and two experimental groups which were intraperitoneally injected with 200 （n=15）and 500 mg/kg ZnO（n=15） every other day for 5 times. Control group （n=15）was injected equal volume saline. After one week of treatment, the coefficients of organs and serum biochemical parameters, gonadal hormone （T and E2） were measured, respectively. Main organs were observed in pathology. Sperm count, sperm motility rate, abnormal sperm rate were observed under microscope. The distribution of nano-particles in liver and testis were observed by Electron microscopy. Germ cell apoptosis was evaluated by TUNEL. Results Compared with the control group, the organ coefficient were not significantly changed after lower-dose（200mg/kg） nano-ZnO administration, but the higher dose （500mg/kg） nano-ZnO administrated mice show significantly elevated quality coefficient of spleen and kidney （P〈0.05）. In low-dose group, serum ALT、TBIL、CHO、UA、CK were significantly changed （P〈0.05）. In contrast, the high-dose treated mice showed significant changed levels of serum ALT、AST、TBA、AKP、TG、CHO （P〈0.05） indicating the liver damage. Moreover, serum BUN、Cr and UA increased obviously, suggesting the kidney damage. In both of these two administrations, the levels of serum testosterone （T）and estradiol （E2） were equivalent with the control （P〉0.05）. The observation of reproductive function found that the decreased sperm count and function in both of two treatments. Meanwhile, the results of TURNEL assay showed that the apoptosis of testicular germ cell increased especially in high-dose group with the significant change of apoptosis index （P〈0.05）. Edema and degeneration in hepatocytes of the mice exposed to high-dose zinc oxide were found. Renal pathological changes and inflammatory cells infiltration in pancreas interstice were obvious. Besides the above organic tissues, no other significant histopathological changes were found in the heart, testis and epididymis. Nano-ZnO were gathered in lysosomes and mitochondria of the liver cells and testicular cells, which can be found by Electron microscopy. Conclusion The low dose of nano-ZnO may have mild effects on the mouse liver, kidney and heart function, moreover, the live sperm rate and the rate of sperm deformity were affected in this group. For high dose of nano-ZnO treated mice, the damage on the liver, kidney and heart were obvious. Especially, the reproductive system was damaged with the decreased sperm count and function as well as testicular germ cell apoptosis.
出处
《医学研究生学报》
CAS
2010年第4期357-364,共8页
Journal of Medical Postgraduates
基金
江苏省卫生厅开放课题(XK02200901-NG09)
关键词
纳米颗粒
氧化锌
雄性小鼠
生殖系统
细胞凋亡
Nanoparticle
Zinc oxide
Male mouse
Reproductive system
Cell apoptosis
