孕期暴露DEHP对子代雌性大鼠下丘脑神经内分泌基因表达的影响

Effect of DEHP exposure during pregnancy on hypothalamic neuroendocrine gene expression in offspring female rats

目的 探究孕期DEHP暴露对后代下丘脑生殖和能量代谢相关神经内分泌基因的影响。方法 将16只受孕成功的SD大鼠随机分为溶剂对照组(D0)和DEHP剂量500 mg/kg组(D500),从妊娠第1天开始连续灌胃18 d,1次/d。F1代子鼠出生后第1天定为PND0,记录新生鼠体质量、死胎、畸胎情况。F1代子鼠于28日龄时处死,采用荧光定量PCR技术检测F1代雌鼠下丘脑生殖相关基因(Kiss1、GnRH)和能量代谢相关基因(Npy、Agrp、Pomc)的mRNA表达。同时,采用下丘脑冷冻切片免疫荧光技术分析上述基因的蛋白表达水平。结果 孕期接触DEHP可导致新生鼠体质量下降,下丘脑GnRH mRNA表达增加,腹前室旁核(AVPV)区Kiss1基因及其编码蛋白Kisspeptin表达增加。在下丘脑弓状核中,Kiss1和Npy基因的mRNA和蛋白表达无明显差异,Agrp基因mRNA和蛋白表达量显著增加,Pomc基因mRNA和蛋白表达量显著降低。结论 孕期DEHP暴露可影响子代雌性大鼠下丘脑生殖和能量代谢相关基因的转录和蛋白表达。

Objective To investigate the effect of DEHP exposure during pregnancy on hypothalamic reproduction and energy metabolism related neuroendocrine genes in offspring.Methods Sixteen pregnant SD rats were randomly divided into solvent control group(D0) and 500 mg/kg DEHP group(D500),which were given continuous gavage for 18 days from the first day of pregnancy,once a day.PND0 was set as the first day after birth of F1 generation offspring mice,and the body mass,litter size and sex ratio of newborn mice were recorded,as well as the existence of stillbirth and teratogenesis.The mice were sacrificed at 28 days of age,and the mRNA expression of reproductive genes(Kiss1,GnRH) and energy metabolism-related genes(Npy,Agrp,Pomc) in the hypothalamus of the offspring of female mice was detected by fluorescence quantitative PCR.Meanwhile,immunofluorescence technique was used to analyze the protein expression levels of the above genes.Results Exposure to DEHP during pregnancy resulted in body mass loss in neonatal mice.GnRH mRNA expression increased in hypothalamus,Kiss1 gene and its encoding protein kisspeptin expression increased in AVPV region.mRNA and protein expression of Kiss1 and Npy genes were not significantly different in the hypothalamic arcuate nucleus.The mRNA and protein expression of Agrp gene were significantly increased,while the mRNA and protein expression of Pomc gene were significantly decreased.Conclusion DEHP exposure during pregnancy can affect the transcription and protein expression of hypothalamic endocrine genes related to reproductive and energy metabolism in offspring female rats.