纳米氧化镍亚慢性染毒致雄性SD大鼠生殖毒性机制  

Mechanismof reproductive toxicity of sub-chronic exposure to nickel oxide nanoparticles in male SD rats

作　　者：范兴君[1] 谷红梅[1] 杨勇[1] 于凤波[2,3] 陈琦 由丽梅 高金霞[1] 牛莹莹[1] 刘凤海[1] FAN Xing-jun;GU Hong-mei;YANG Yong;YU Feng-bo;CHEN Qi;YOU Li-mei;GAO Jin-xia;NIU Ying-ying;LIU Feng-hai(School of Public Health,MuDanJiang Medical Colleage,MuDanJiang,Heilongjiang 157011,China;不详)

机构地区：[1]牡丹江医学院公共卫生学院,黑龙江牡丹江157011 [2]牡丹江医学院药学院,黑龙江牡丹江157011 [3]广东省高州市高州人民医院,广东高州525200

出　　处：《中国职业医学》2022年第2期140-147,共8页China Occupational Medicine

基　　金：黑龙江省省属高校基本科研业务项目生物医药类支持计划项目(2020-KYYWFMY-0744)。

摘　　要：目的探讨纳米氧化镍(Nano NiO)亚慢性染毒对雄性SD大鼠内分泌功能的影响,并分析其对睾丸细胞毒作用与机制。方法将无特定病原体级雄性SD大鼠随机分为5组,每组10只。采用非暴露式气管滴注法染毒,低、中、高剂量组大鼠分别予质量浓度为0.16、0.80、4.00 g/L的Nano NiO混悬液,空白对照组大鼠予等体积0.9%氯化钠溶液,阳性对照组大鼠予质量浓度为4.00 g/L的微米氧化镍混悬液,每3天滴注1次,持续9周。染毒结束后,采用原子荧光光谱法测定血液和睾丸组织中镍的水平,采用酶联免疫吸附实验检测血清性激素水平,采用流式细胞术分析睾丸细胞倍体比值、细胞周期和细胞凋亡率,采用蛋白质印迹法检测睾丸组织中凋亡相关蛋白相对表达水平。结果阳性对照组与3个剂量组大鼠血液和睾丸组织中镍水平均高于空白对照组(P值均<0.05);中、高剂量组大鼠血液和睾丸组织中镍水平均高于阳性对照组(P值均<0.05);血液与睾丸组织中镍水平呈正相关(P<0.01)。中、高剂量组大鼠血清睾酮、卵泡刺激素(FSH)和黄体生成素(LH)水平均低于空白对照组(P值均<0.05);但各组大鼠血清促性腺激素释放激素水平比较,差异无统计学意义(P>0.05)。与空白对照组比较,中、高剂量组睾丸生殖细胞单倍体、二倍体细胞构成比和G0/G1期、S期细胞比值均下降(P值均<0.05),睾丸细胞四倍体细胞构成比、G2/M期细胞比值和早期凋亡率均增加(P值均<0.05)。与空白对照组比较,中、高剂量组大鼠睾丸组织中B淋巴细胞瘤-2(BCL-2)蛋白相对表达水平和BCL-2/BCL-2相关X蛋白(BAX)比值均下降(P值均<0.05),BAX和半胱氨酸天冬氨酸蛋白酶-3蛋白相对表达水平均升高(P值均<0.05)。与阳性对照组比较,高剂量组大鼠血液和睾丸组织中镍水平均升高(P值均<0.05),睾丸细胞单倍体构成比和G0/G1期、S期比值以及睾丸组织中BCL2/BAX比值均下降(P值均<0.05),�Objective endocrine function of male SD rats,and to analyze the toxicity and mechanism of Nano NiO on testicular cells.Methods The specific pathogens free male SD rats were randomly divided into five groups with ten rats in each group.Rats in low-,mediumand high-dose groups were given Nano NiO suspension with the mass concentration of 0.16,0.80 and 4.00 g/L,respectively;rats in blank control group were given equal volume of 0.9%sodium chloride solution;rats in positive control group were given micron nickel oxide suspension with the mass concentration of 4.00 g/L.Drip every three days for nine weeks.After the Nano NiO exposure,atomic fluorescence spectrometry was used to determine the levels of nickel in the blood and testicular tissue.The enzyme-linked immunosorbent assay was used to detect serum level of sex hormone.The ploidy ratio,cell cycle and apoptosis rate of testicular cells were analyzed by flow cytometry.Western blotting was used to detect the relative expression of apoptosisrelated proteins in the testis.Results The level of nickel in blood and testicular tissue of rats in positive control group and the three doses groups were higher than that of blank control group(all P<0.05).The level of nickel in blood and testicular tissue of rats in the medium-dose and high-dose groups were higher than that in the positive control group(all P<0.05).There was a positive correlation between the level of nickel in blood and testicular tissue(P<0.01).The serum levels of testosterone,folliclestimulating hormone(FSH)and luteinizing hormone(LH)in the medium-and high-dose groups were lower than that in blank control group(all P<0.05).However,there was no significant difference in serum gonadotropin-releasing hormone among all groups(P>0.05).Compared with the blank control group,the proportion of haploid and diploid cells and the ratio of cells in G0/G1 and S phase decreased in the medium-and high-dose groups(all P<0.05),the tetraploid cell ratio,G2/M cell ratio and early apoptotic rate of testicular cells increased(all

关 键 词：纳米氧化镍 睾丸 性激素 B淋巴细胞瘤-2(BCL-2) BCL-2相关X蛋白 细胞周期 细胞凋亡 大鼠 

分 类 号：R134.3[医药卫生—劳动卫生]