硫酸镍对体外培养大鼠睾丸间质细胞超微结构的影响  

Effects of nickel sulfate on ultramicrostructure alterations of rat leydig cells in vitro

作　　者：郑菁[1] 张志刚[1] 孙应彪[1] 苏莉[1] 夏胡[1] 李成云[1] 张瑞[1] 李晋[1] 

机构地区：[1]兰州大学公共卫生学院,甘肃兰州730000

出　　处：《工业卫生与职业病》2012年第1期29-32,共4页Industrial Health and Occupational Diseases

基　　金：中央高校基本科研业务费专项资金(Lzujbky-2009-89)

摘　　要：目的通过观察硫酸镍(NiSO4)对体外培养大鼠睾丸间质细胞超微结构的改变,初步探讨镍致睾丸间质细胞损伤的特点。方法采用胶原酶消化、Percoll分离液密度梯度离心和差速贴壁法体外分离、纯化睾丸间质细胞,采用3β-HSD染色法鉴定其纯度;采用MTT法观察NiSO4(浓度为0.0、62.5、125.0、250.0、500.0和1000.0μmol/L)染毒3、6和12h后,对睾丸间质细胞生长抑制的影响;采用透射电镜观察染毒6h后大鼠睾丸间质细胞超微结构的变化。结果纯化的睾丸间质细胞采用3β-HSD染色后,镜下可见睾丸间质细胞被染成蓝黑色,细胞纯度可达到95%以上;与对照组比较,染毒6h,NiSO4浓度为125.0μmol/L及其以上浓度组对间质细胞生长具有明显的抑制作用,差异有统计学意义(P<0.05或P<0.01);NiSO4可引起大鼠睾丸间质细胞线粒体和内质网超微结构改变,主要表现为线粒体肿胀,滑面内质网扩张及脱颗粒现象,溶酶体增多,尤其NiSO4浓度为500.0和1000.0μmol/L变化明显。结论 NiSO4可引起大鼠睾丸间质细胞超微结构改变。Objective By observing the ultramicrostructure alterations of leydig cells to investigate the injury features of leydig cells induced by nickel sulfate（NiSO4 ）in vitro. Methods Leydig cells were isolated using collagenase dispersion. The density gradient centrifugation with Percoll separating medium and differential adhesion assay were used to purify leydig cells, then the leydig cells were identified with 3β-HSD staining; Leydig cells of exponential growth phase were treated with different concentrations of nickel sulfate at doses of 0. 0, 62.5, 125.0, 250.0, 500.0 and 1000.0 μmol/L, and the proliferation of leydig cells was detected with MTT chromatography after separately exposed for 3 h, 6 h and 12 h. Transmission electron microscopy（TEM）was used to observe the ultramicrostrueture changes of leydig cells after exposed for 6 h. Results The cytoplasm of leydig cells were stained specifically in blue black with 3β-HSD staining, and the purity of leydig cells was above 95%; Compared with the control group, 125.0μmol /L and above concentrations could significantly inhibit the growth of leydig cells （ P 〈0.05 or P〈 0.01 ） ; The various abnormal changes of mitochodria and endoplasmic reticulum were observed with TEM including mitochondrial swelling, ectasia and degranulation of smooth endoplasmic reticulum and increased lysomal in NiSO4 500.0μmol/L and 1000.0 μmol/L group especially. Conclusions NiSO4 can induce the uhramicrostructure alterations of rat leydig cells.

关 键 词：硫酸镍 睾丸间质细胞 超微结构 

分 类 号：R114[医药卫生—卫生毒理学]