摘要
目的探讨邻苯二甲酸二(2-乙基己基)酯(DEHP)对类固醇合成急性调节蛋白(St AR)基因表达水平和蛋白表达水平的影响。方法用不同剂量DEHP(0.05~0.80 mmol/L)对MCF-7细胞染毒24、48 h,采用荧光定量PCR检测St AR基因表达水平,观察DEHP染毒的剂量-效应关系;用单一剂量DEHP 0.80 mmol/L染毒细胞3、6、12、24、48 h,荧光定量PCR检测St AR基因表达水平,观察DEHP染毒的时间-效应关系;DEHP(0.10~0.80)mmol/L染毒24 h,western blot检测St AR蛋白表达水平的改变。结果 DEHP染毒24 h,(0.10~0.80)mmol/L剂量组St AR基因表达水平都显著高于对照组50%~130%,差异有统计学意义(P<0.01);DEHP染毒48 h,(0.05~0.80)mmol/L剂量组St AR基因表达水平比对照组升高30%~185%,差异有统计学意义(P<0.01或P<0.05)。DEHP 0.8 mmol/L染毒3 h后St AR基因表达比对照组升高35%(P<0.05),染毒6 h后St AR基因表达升高65%(P<0.01),染毒12 h升高130%(P<0.01),染毒24 h升高145%(P<0.01),染毒48 h升高260%(P<0.01)。Western blot结果显示,DEHP 0.10 mmol/L组St AR蛋白表达与对照组比较无明显变化,0.20 mmol/L组St AR蛋白表达比对照组明显升高35%(P<0.05),0.40 mmol/L组St AR蛋白表达升高55%(P<0.01),0.80 mmol/L组St AR蛋白表达升高70%(P<0.01)。结论 DEHP可引起St AR基因表达和蛋白表达水平上调,推测DEHP可能通过影响类固醇激素合成过程中St AR表达水平变化,从而产生生殖毒性作用。
[Objective] To study the effects of di（2-ethylhexyl） phthalate（DEHP） on the expression of steroidogenio acute regulatory protein（St AR）.[Methods]MCF-7 cells were treated with various doses of DEHP（0.05-0.80 mmol/L） for 24 hours and 48 hours. The expression of St AR gene was detected by real-time PCR,and the dose-effect relationship was observed.Additionally,the cells were treated by a single dose of DEHP at 0.8 mmol/L for 3 hours,6 hours,12 hours,24 hours and 48 hours,the expression of St AR gene was detected by real-time PCR,and the dose-effect relationship was observed. The cells were treated by DEHP at 0.10-0.80 mmol/L for 24 hours,and the expression of St AR protein was detected by western blot.[Results]After exposure to DEHP for 24 hours,the expression level of St AR gene in 0.10-0.80 mmol/L DEHP groups was30%-185% higher than that in the control group,and the differences were statistically significant（P〈0.01）. After exposure to DEHP for 48 hours,the expression level of St AR gene in 0.05-0.80 mmol/L DEHP groups was 30%-185% higher than that in the control group,and the differences were statistically significant（P〈0.01,P〈0.05）. In 0.80 mmol/L DEHP group,the expression level of St AR gene after 3 hours increased by 35% as compared with the control group（P〈0.05）,the expression level after6 hours increased by 65%（P〈0.01）,the expression level after 12 hours increased by 130%（P〈0.01）,the expression level after24 hours increased by 145%（P〈0.01）,and the expression level after 48 hours increased by 260%（P〈0.01）. The results of Western blot showed that there was no statistically significant difference in the expression levels of St AR protein between 0.10 mmol/L DEHP group and the control group,while the expression level of St AR protein in 0.20 mmol/L DEHP group increased by 35% as compared with the control group（P〈0.05）,the expression level in 0.40 mmol/L DEHP group increased by 55%（P〈0.01）,and the expression level in 0.80 mmol/L DEHP group increased by 70%（P〈0.01）.[Conclusion]DEHP could induce up-regulation of St AR gene expression and protein expression,and these findings indicated that DEHP probably induce reproductive toxicity through affecting the metabolism of steroidogenio acute regulatory protein during steroid hormone synthesis.
作者
彭鹏
王利
徐新云
袁建辉
黄海燕
PENG Peng;WANG Li;XU Xin-yun;YUAN Jian-hui;HUANG Hai-yan(School of Life Science, Shenzhen University, Shenzhen Guangdong,518060, China;1nstitute of Toxicology, Shenzhen Center for Disease Control and Prevention, Shenzhen Guangdong,518055,China;School of Public Health,South China University, Hengyang Hunan, 421001, China)
出处
《职业与健康》
CAS
2017年第24期3340-3343,共4页
Occupation and Health
基金
深圳市科技研发基础研究项目(JCYJ20170306160553495)
