The Effects Of Metabolic And Endocrine Alterations On Uterine Endometrium And Offspring Metabolism

Part?:Hypertriglyceridemia in female rats during pregnancy induces obesity in male offspring via altering hypothalamic leptin signalingBackground:Obesity is risk factor for mother and child health.Maternal obesity influences the child's long-term development.Obesity may be genetic,epigenetics and may be due to alteration in hypothalamic feeding circuits.Though,the mechanism concerned in this process is still uncertain.In the present study,we proposed that overfeeding of a high-fat diet during pregnancy in female rats altered metabolic phenotypes in F1 generation and might be the contribution of hypothalamic leptin signaling.Methods:Female rats were purchased and housed for one week.When the rats were get pregnant,then provided high fat diet.At 21 days of age,offspring were scarified for hypothalamic immunofluorescence and western blot.For leptin sensitivity,leptin responsiveness and the number of immunopositive neurons for phosphorylated signal transducer and activator transcription 3?pSTAT3?were analyzed.Neuropeptide Y in the arcuate nucleus of the hypothalamus and in nucleus tractus solitaries was examined.Results:Triglycerides and leptin levels were increased in the high-fat diet mother as compared with control mother.The number of neuropeptide Y positive cell bodies and neurons was significantly increased in the high-fat diet-F1 offspring?HDF-F1?as compared to Chow-F1.Leptin administration significantly decreased the food intake and increased the pSTAT3 expression levels in neurons in the arcuate nucleus of Chow-F1,while there is no significant increase in the HFD-F1 group.Conclusions:From the present domino effect,we conclude that mothers exposed to high-fat diet during pregnancy may pass the obese phenotype to the succeeding generation via altering hypothalamic leptin signaling.Triglycerides acts centerly via their receptor in Arc and manipulate the feeding circuit genes.Part ?:Androgen-induced alterations in endometrial proteins crucial in recurrent miscarriagesBackground:Endometrium carries androgen receptors.High androgen level impairs endometrial receptivity in women experiences the recurrent miscarriage and implantation failure.The mechanism of androgen actions on endometrium is still uncertain.We hypothesized that androgen has a direct effect on the endometrium in women with recurrent miscarriage.Methods:In the present study,we assess the impact of androgen?A₂?at high concentration(10^-7 M)on Ishikawa cells compared with the physiological concentration of androgen(10^-9 M).To go into deeper analysis,we use global stable isotopes labeled profiling tactic using iTRAQ reagents,followed by 2D LC-MS/MS.Western blot was performed for iTRAQ results validation.Gene for CDKN2a was blocked with the specific siRNA.Results:We determine 175 non-redundant proteins,and 18 of these were quantified.The analysis of differentially expressed proteins?DEPs?identified 8 up-regulated proteins and 10 down-regulated in the high androgen group.These DEPs were examined by ingenuity pathway?IPA?analysis and established that these proteins might play vital roles in recurrent miscarriage and endometrium receptivity.In addition,proteins cyclin-dependent kinase inhibitor 2a?CDKN2a?,endothelial protein C receptor?EPCR?,armadillo repeat for velocardiofacial?ARVCF?were independently confirmed using western blot.Knockdown of CDKN2a significantly decreased the expression level of CDKN2a protein in ishikawa cells,and decreased migration?p<0.01?,invasion?p<0.05?,proliferation?p<0.05?,and the rate of Jar spheroid attachment?p<0.05?to Ishikawa cell monolayer.Conclusions:The present results suggest that androgen at high concentration could alter the expression levels of proteins related to endometrium development and embryo implantation,which might be a cause of the impaired endometrial receptivity and miscarriage.