| Objective:To investigate the effects and the mechanism of prenatal DEHP exposure on the food intake and energy metabolism of offspring rats,the leptin levels and expression of genes and proteins related to food intake and energy metabolism in the hypothalamus ARC region were detected.Method:64 healthy adult SPF pregnant SD rats were randomly divided into 4 groups according to body weight: the control(corn oil)group and the 2,10,and 50 mg/kg DEHP exposure group.In G14 ～ G19,the DEHP was applied by gavage.The offspring were weaned 21 days after birth and reared into male and female cages until 42 and 70 days of age.Female and male were randomly selected for each age group.The blood was taken after femoral artery,The cervical spine was dislocated and sacrificed.The fresh brains were taken.The ARC area was taken according to the anatomical mapping of the rat brain.The expression levels of energy metabolism related genes in the thalamus ARC area were determined by the 48 gene chip RT2 Profiler PCR Array designed by ourselves to perform real-time quantitative PCR.Male and female(8 in each group,32 in total)were taken from each age group.The whole brains were taken after perfusion fixation of cardiac paraformaldehyde.Automatic oscillating microtome was used to cut to 40μm slices.The expression levels of energy metabolism related proteins NPY and POMC in the ARC area of the hypothalamus were determined by use immunofluorescence histochemistry.Serum leptin levels were determined by Luminex liquid chip technology.Result:(1)The weights of male offspring rats in PND21,PND28,PND35,PND42,PND49 were statistically significant.Compared with the control,the weight of PND21,in the 2 mg/kg DEHP group was reduced(P<0.05).the weight from PND21 to PND49 in the 10 mg / kg DEHP dose group was reduced(P<0.05).The weight of PND21,PND28,PND35 and PND49 in 50mg/kg DEHP dose group was reduced(P<0.05).The weight of PND21,PND49,and PND56 in female offspring rats was statistically significant.Compared with the control,the weight of PND49 and PND56 in the 2 mg / kg DEHP dose group was induced(P<0.05).The weight of PND21 in the 10 mg / kg DEHP dose group increased(P<0.05),and PND49 in the 10 mg / kg DEHP dose group increased(P<0.05).the weight of PND21 in the 50 mg / kg DEHP dose group decreased(P<0.05).(2)The food intake of male in PND35 was statistically significant.Compared with the control,the food intake of the 10 mg / kg DEHP dose group increased(P<0.05).The food intake of female in PND35 and PND63 had statistically significant differences.Compared with the control,food intake in PND35 and PND63 increased in the 2mg / kg DEHP dose group(P<0.05).(3)There was significant difference in serum Leptin levels between female adolescent rats among the various DEHP dose groups(P<0.05).Compared with the control group,the serum Leptin levels of the 10 mg / kg dose group decreased(P<0.01).There was significant difference in serum Leptin levels between male adult rats among the various DEHP dose groups(P<0.05).Compared with the control,the serum Leptin levels in the 10 mg/kg dose group decreased(P<0.05).(4)For adolescent male rats,the Cntfr gene expression level increased in the 2mg/kg DEHP group(P<0.05).The Cnr1 gene expression level of the 2mg/kg DEHP dose group decreased(P<0.05).The expression level of Hcrt increased in the 10 mg/kg DEHP group(P<0.05).The expression level of Insr,Atrn,Ghrl gene decreased in the 10mg/kg DEHP group(P<0.05).The expression level of Lepr decreased in the 50 mg/kg DEHP group(P<0.05).The expression level of Nmbr decreased in the 2 mg/kg and 10 mg/kg DEHP group(P<0.05).For adolescent female,rats the expression level of Cartpt decreased in the 2 and 10 mg/kg DEHP groups(P<0.05).The expression level of Crh increased in the 10 mg/kg DEHP group(P<0.05).The expression levels of Npy and Ghsr in the 10mg/kg DEHP group increased(P<0.05).The expression level of Adipor2 decreased in the 50 mg/kg DEHP group(P<0.05).For adult male rats,the expression levels of Npy decreased in the 10 mg/kg and 50 mg/kg DEHP groups(P<0.05).The expression levels of Pomc decreased in the 2,10,and 50 mg/kg DEHP groups(P <0.05).The expression level of Prl increased in the 50 mg/kg DEHP dose group(P <0.05).The expression level of Lepr increased in the 50 mg/kg DEHP group(P<0.05).The expressions of Gal and Cartpt in adult female rats were statistically significant.Compared with the control,the expression levels of Gal in the 50 mg/kg DEHP group increased(P<0.05).The expressions of Cartpt in the 50 mg/kg DEHP group increased(P<0.05).(5)The expression of NPY protein in the hypothalamic ARC region of adolescent male was statistically significant.Compared with the control,the expression of NPY in the 50 mg/kg DEHP group increased(P<0.05).The expression of NPY protein of female was statistically significant.Compared with the control,the expression of NPY in the 2 mg/kg DEHP group increased(P<0.05).The expression of POMC protein of adolescent male rats was statistically significant.Compared with the control,the relative expression of POMC in the 2 mg/kg DEHP dose group decreased(P<0.05).There was no significant difference in POMC protein of female.The expression of NPY protein of adult male rats was statistically significant.Compared with the control,the expression of NPY in the 10 mg/kg DEHP group increased(P<0.05).There was no significant difference in the expression of NPY protein in female rats.The expression of POMC protein adult male rats was statistically significant.Compared with the control,the expression of POMC in the 10 mg/kg DEHP group decreased(P<0.05).There was no significant difference in the relative expression of POMC protein of female.Conclusion:(1)Prenatal DEHP exposure can affect the growth and development of offspring rats;(2)Prenatal DEHP exposure can affect serum leptin levels in offspring;(3)Prenatal DEHP exposure can affect food intake and energy metabolism-related genes and Protein expression;(4)Prenatal DEHP exposure can affect feeding and energy metabolism in offspring rats,and the related mechanisms need to be further studied. |
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