邻苯二甲酸单乙基己基酯对原代培养新生雌性大鼠下丘脑神经元生殖神经内分泌相关基因表达影响

Effects of MEHP on expression of reproductive neuroendocrine related genes in primary cultured hypothalamus neurons of neonatal female rats

目的探讨邻苯二甲酸单乙基己基酯(mono-2-ethylhexyl phthalate,MEHP)对新生雌性大鼠下丘脑生殖神经内分泌功能的影响。方法对出生24h内的新生雌性SD大鼠下丘脑神经元进行原代培养,分别暴露于含终浓度为0(对照)、10^(-15)、10^(-12)、10^(-9)、10^(-6)mol/L MEHP的培养基染毒24h。采用qRT-PCR法检测Gn RH、Gn RHr、Kiss1及Kiss1r基因的表达水平。结果与对照组相比,各浓度MEHP暴露组原代培养新生雌性大鼠下丘脑神经元Kiss1基因的表达水平均较低,差异有统计学意义(P<0.05);而Gn RH、Gn RHr和Kiss1r基因的表达水平均无明显改变。结论 MEHP有可能通过下调新生雌性大鼠下丘脑神经元Kiss1基因表达水平来干扰生殖神经的内分泌调控功能。

Objective To study the impact of mono-2-ethylhexyl phthalate （MEHP） exposure on the expression of reproductive neuroendocrine regulation related genes in primary cultured hypothalamic neurons of neonatal female rats. Methods The hypothalamus of neonatal Sprague Dawley rats were collected within 24 h and cultured for 6 days before 24 h MEHP treatment with different doses （0 mol/L, 10-15 mol/L, 10-12 mol/L, 10-9 mol/L and 10-6 mol/L respectively）. Total RNA was extracted by using TRIzol following the manufacturer＇s protocol, qRT-PCR was used to determine the expressions of reproductive neuroendocrine-related genes：GnRH, GnRHr, Kissl and Kisslr. Results MEHP treatment did not change the expressions of GnRH, GnRHr and Kisslr genes in primary cultured hypothalamic neurons. Compared with control group,the Kissl genes expression was significantly down-regulated in 10-j5 mol/L, 10-12 mol/L, 10-9 mol/L and 104 mol/L MEHP treatment groups. Conclusion MEHP exposure may change the Kissl expressions of primary cultured hypothalamic neurons of female neonatal rats. Neonatal MEHP exposure may impair reproductive neurocndocrine system by altering Kissl expression in hypothalamus.