摘要
目的观察ATM蛋白激酶-检测点激酶2(ATM-Chk2)通路在氟抑制大鼠曲细精管上皮支持细胞增殖中的作用。方法将18日龄SPF级雄性SD大鼠曲细精管上皮支持细胞分别暴露于含0(对照)、0.25、0.50、1.00、2.00、4.00 mmol/L氟化钠的血清培养基。分别于染毒24、48、72 h,采用MTT法测定支持细胞的增殖情况;于染毒24 h,采用实时荧光定量PCR技术检测支持细胞内ATM、Chk2 m RNA的表达水平。结果与对照组比较,染毒24、48、72 h时,4.00 mmol/L氟化钠染毒组大鼠曲细精管上皮支持细胞的抑制率均较高,差异有统计学意义(P<0.05);且随着氟化钠染毒剂量的升高和染毒时间的延长,氟对曲细精管上皮支持细胞的抑制率均呈上升趋势。与对照组比较,1、2 mmol/L氟化钠染毒组曲细精管上皮支持细胞ATM和Chk2 m RNA的表达水平均升高,而4 mmol/L氟化钠染毒组ATM和Chk2 m RNA的表达水平均降低,差异有统计学意义(P<0.05);且随着氟化钠染毒剂量的升高,曲细精管上皮支持细胞ATM、Chk2 m RNA的表达水平均呈下降趋势。结论不同剂量氟抑制生殖细胞增殖可能是通过干扰曲细精管上皮支持细胞中DNA损伤检验点ATM、Chk2的表达而实现的。
Objective To explore the role of ATM kinase/checkpoint kinase 2 pathway in inhibiting proliferation of epithelial Sertoli cell in the seminiferous tubules of rats exposed to fluoride. Methods The Sertoli ceils in seminiferous tubules were isolated from !8-day-old SPF male SD rats and were exposed to serum medium containing sodium fluoride at the doses of 0,0.25,0.50,1.00,2.00 and 4.00 mmol/L, respectively. After 24,48,72 h of exposure repeetively, the proliferation of Sertoli cell was detected by MTT method. After 24 h of exposure, mRNA relative expression levels of A TM and Chk2 gene were detected by real-time RT-PCR technique. Results Compared with the control group, sodium fluoride at the dose of 4.00 mmol/L could significantly inhibit the proliferation of Sertoli cells after 24,48 and 72 h of exposure respectively. The inhibition effect of fluoride on the Sertoli cell proliferation showed a rising trend with the increase of exposure dosage and time. Compared with the control group,the mRNA expression levels of ATM and Chk2 gene in 1 and 2 mmol/L fluoride exposure groups significantly increased but decreased significantly in 4 mmol/L fluoride exposure group, and the relative expressions of the two gene in all exposure groups presented a downward trend. Conclusion The proliferation inhibition of fluoride on Sertoli cell may be closely related with interference of ATbl and Chk2 expression.
出处
《环境与健康杂志》
CAS
北大核心
2015年第1期29-32,共4页
Journal of Environment and Health
基金
2013年地方高校国家级大学生创新训练计划项目(201310573008)
关键词
氟
曲细精管上皮支持细胞
增殖
ATM蛋白激酶
检测点激酶2
Fluoride
Epithelial Sertol
cell in the seminiferous tubules
Proliferation
ATM kinase
Checkpoint kinase 2（Chk2）
