Li T Z, Gong F, Zhang B Y, Sun J D, Zhang T, Kong L, Xue Y Y, Tang M
Key Laboratory of Environmental Medicine and Engineering, Ministry of Education, School of Public Health & Collaborative Innovation Center of Suzhou Nano Science and Technology, Southeast University, Jiangsu Key Laboratory for Biomaterials and Devices, Nanjing 210009, China.
Zhonghua Shao Shang Za Zhi. 2016 Oct 20;32(10):606-612. doi: 10.3760/cma.j.issn.1009-2587.2016.10.007.
To explore the acute toxic effect and the cumulative target organ of silver nitrate and nano-silver with two different particle diameters in rats. Thirty-six adult SD rats were divided into small particle size nano-silver group (SNS), large particle size nano-silver group (LNS), silver nitrate group (SN), and control group (C) according to the random number table, with 9 rats in each group. The rats of the four groups were respectively injected with 10 mg/mL nano-silver solution (particle diameter of 20 nm, prepared by saline) in silver dose of 30 mg/kg by tail vein for once, 10 mg/mL nano-silver solution (particle diameter of 100 nm, prepared by saline) in silver dose of 30 mg/kg, 1.67 mg/mL silver nitrate solution (prepared by glucose solution) in silver dose of 3 mg/kg, and 30 mg/mL polyvinylpyrrolidone solution (prepared by saline) in dose of 90 mg/kg. (1) Toxicity test. The general observation was performed within 14 days after injection, and the deviation between value of body mass before injection and each of that on post injection day (PID) 1, 7, and 14 were respectively recorded. On PID 1, 7, and 14, 3 rats of each group were harvested for determination of serum content of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein, and albumin by fully automatic biochemical analyzer. Then the rats were sacrificed immediately, and heart tissue, liver tissue, spleen tissue, lung tissue, kidney tissue, and brain tissue were collected to calculate the organ coefficient. Organ samples with obvious changes in organ coefficient were collected for histopathological observation by HE staining, with 3 samples in each group at each time point. (2) Bio-distribution. The specimens of heart, liver, spleen, lung, and kidney of rats from groups SNS, LNS, and SN were collected for detection of silver content by inductively coupled plasma mass spectrometry, with 3 samples in each group at each time point. Data were processed with analysis of variance of factorial design, LSD test, and Dunnett's T3 test. (1) The general condition of rats in groups C and SN after injection were normal. The state of rats of groups SNS and LNS was poor with black secretion in the eye and other phenomena on PID 1, which recovered from PID 3 on. (2) The deviations between values of body mass before injection and that on PID 14 in rats of groups LNS and SN were significantly decreased as compared with deviation of group C (with values below 0.01), but deviation of group SNS was not significantly changed (>0.05). The deviations between values of body mass before injection and each of that on PID 1 and 7 in rats in the other three groups were similar to those in group C (with values above 0.05). (3) Compared with those in group C, the serum content of total protein of rats in group SN on PID 1 was significantly decreased, and liver coefficient was significantly increased (with values below 0.05). On PID 1, the serum content of ALT of rats in group LNS was (61.0±8.7) U/L, which was significantly higher than that in group C [(40.0±4.6) U/L, <0.01]. Compared with those in group C [(126.0±3.5) U/L and 4.05±0.23], the serum content of AST of rats in groups SNS and LNS on PID 1[(249.7±107.2) and (237.0±38.3) U/L] was significantly increased, and liver coefficients (3.50±0.38 and 3.31±0.07) were significantly decreased, with values below 0.05. Compared with those in group C [(69.2±4.8) U/L and 4.32±0.39], the serum content of AST of rats in groups SNS and LNS on PID 7 [(181.0±51.5) and (167.7±16.5) U/L] was also significantly increased, and liver coefficients (3.55±0.18 and 3.62±0.21) were also significantly decreased, <0.05 or <0.01. On PID 14, the four liver biochemical indexes in serum and all organ coefficients of rats in the other three groups were similar to those in group C (with values above 0.05). (4) The liver of rats in group SN had slight degeneration on PID 1, the liver cells around the central vein of liver of rats in group SNS had slight degeneration on PID 7, and the liver cells got severely eosinophilic degeneration in liver of rats in group LNS on PID 7. There was no significant pathological change in the liver of rats in each group at the rest time points. (5) The silver content of lung and kidney in rats of group SNS on PID 1, that of spleen and kidney in rats of group LNS on PID 1, and that of heart and kidney in rats of groups LNS and SNS on PID 7 was significantly less than that of group SN (with values below 0.05). The silver content of liver and spleen in rats of group SNS on PID 14 was significantly more than that of group SN (with values below 0.05). Compared with that of group SN, the silver content of lung on PID 1 and liver on PID 7 in rats of group LNS was significantly increased (with values below 0.05). On PID 14, there was no significant change in the silver content of all organs of rats between group SN and group LNS (with values above 0.05). The silver content of heart, lung, and kidney on PID 1 and heart on PID 7 in rats of group LNS was significantly more than that of group SNS (with values below 0.05). On PID 14, the silver content of each organ of rats in group SNS was close to that in group LNS (with values above 0.05). Silver nitrate and nano-silver with two different particle diameters have a short acute toxic effect on the liver of rats, and the liver has certain ability of self-healing. Nano-silver is mainly accumulated in the liver. The distribution of nano-silver with large particle diameter in organs is more widely than that of nano-silver with small particle diameter.
探讨硝酸银及两种不同粒径纳米银对大鼠的急性毒性作用及累积靶器官。将36只成年SD大鼠按随机数字表分为小粒径纳米银组（SNS）、大粒径纳米银组（LNS）、硝酸银组（SN）和对照组（C），每组9只。四组大鼠分别经尾静脉注射10 mg/mL纳米银溶液（粒径20 nm，用生理盐水配制），银剂量为30 mg/kg，注射1次；10 mg/mL纳米银溶液（粒径100 nm，用生理盐水配制），银剂量为30 mg/kg；1.67 mg/mL硝酸银溶液（用葡萄糖溶液配制），银剂量为3 mg/kg；30 mg/mL聚乙烯吡咯烷酮溶液（用生理盐水配制），剂量为90 mg/kg。（1）毒性试验。注射后14天内进行一般观察，分别记录注射前体质量值与注射后第1、7、14天体质量值的差值。在注射后第1、7、14天，每组取3只大鼠，用全自动生化分析仪测定血清丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、总蛋白和白蛋白含量。然后立即处死大鼠，取心脏组织、肝脏组织、脾脏组织、肺组织、肾脏组织和脑组织计算脏器系数。对脏器系数有明显变化的脏器样本进行HE染色组织病理学观察，每个时间点每组取3个样本。（2）生物分布。收集SNS组、LNS组和SN组大鼠的心脏、肝脏、脾脏、肺和肾脏标本，用电感耦合等离子体质谱法检测银含量，每个时间点每组取3个样本。数据采用析因设计方差分析、LSD检验和Dunnett's T3检验进行处理。（1）C组和SN组大鼠注射后一般情况正常。SNS组和LNS组大鼠在注射后第1天状态较差，眼部有黑色分泌物等现象，从第3天开始恢复。（2）LNS组和SN组大鼠注射前体质量值与注射后第14天体质量值的差值与C组相比显著降低（P值<0.01），但SNS组差值无显著变化（P>0.05）。其他三组大鼠注射前体质量值与注射后第1天和第7天体质量值的差值与C组相似（P值>0.05）。（3）与C组相比，SN组大鼠注射后第1天血清总蛋白含量显著降低，肝脏系数显著升高（P值<0.05）。注射后第1天，LNS组大鼠血清ALT含量为（61.0±8.7）U/L，显著高于C组[（40.0±4.6）U/L，P<0.01]。与C组[（126.0±3.5）U/L和4.05±0.23]相比，SNS组和LNS组大鼠注射后第1天血清AST含量[（249.7±107.2）和（237.0±38.3）U/L]显著升高，肝脏系数（3.50±0.38和3.31±0.07）显著降低，P值<0.05。与C组[（69.2±4.8）U/L和4.32±0.39]相比，SNS组和LNS组大鼠注射后第7天血清AST含量[（181.0±51.5）和（167.7±16.5）U/L]也显著升高，肝脏系数（3.55±0.18和3.62±0.21）也显著降低，P<0.05或P<0.01。注射后第14天，其他三组大鼠血清中四项肝脏生化指标及所有脏器系数与C组相似（P值>0.05）。（4）SN组大鼠注射后第1天肝脏有轻度变性，SNS组大鼠注射后第7天肝中央静脉周围肝细胞有轻度变性，LNS组大鼠注射后第7天肝细胞出现严重嗜酸性变性。其余时间点各组大鼠肝脏均无明显病理变化。（5）SNS组大鼠注射后第1天肺和肾脏的银含量、LNS组大鼠注射后第1天脾脏和肾脏的银含量、LNS组和SNS组大鼠注射后第7天心和肾脏的银含量均显著低于SN组（P值<0.05）。SNS组大鼠注射后第1天肝脏和脾脏的银含量显著高于SN组（P值<0.05）。与SN组相比，LNS组大鼠注射后第1天肺和注射后第7天肝脏的银含量显著升高（P值<0.05）。注射后第14天，SN组和LNS组大鼠各脏器银含量无显著变化（P值>0.05）。LNS组大鼠注射后第1天心、肺和肾脏的银含量及注射后第7天心的银含量均显著高于SNS组（P值<0.05）。注射后第14天，SNS组大鼠各脏器银含量与LNS组相近（P值>0.05）。硝酸银及两种不同粒径纳米银对大鼠肝脏有短期急性毒性作用，肝脏有一定的自我修复能力。纳米银主要蓄积在肝脏。大粒径纳米银在各脏器中的分布比小粒径纳米银更广泛。
