Quantitative Detection Of The PPARγ In Pregnant Swine Placenta Using FQ-PCR Assay

Objective:To investigate the expression of PPARy in sow placenta at different periods of pregnancy and in different fetal survival state; through the analysis of its correlation with fetal weight, fetal length, the sac length, explore the impact of PPARy on sow placenta and fetal development.Methods:The primers of PPARy and GAPDH for ordinary PCR and FQ-PCR were designed; RNA was extracted from tissue and reverse transcribed into cDNA; target gene fragments were amplified through ordinary PCR, the PCR products were purified and connected to the vector and then transformed into competent cells; PPARy and GAPDH standard plasmid were extracted and used to make FQ-PCR standard curve. Absolute and relative quantification methods of FQ-PCR were used to detect the expression of PPARy in sow placenta at different periods of pregnancy.Results:A FQ-PCR which can be used for absolute and relative quantification of PPARy was established, the detection sensitivity can be up to20copies. During pregnancy25d,40d and70d in the endometrium, the relative expression of PPARy were1.11±0.37,8.65±5.59,6.26±1.93, the change from25d to70d is significant (P<0.01), the change from25d to40d is notable (0.01<P<0.05), the change from25d to70d is not significant. During pregnancy25d,40d and70d in the chorion, the relative expression of PPARy were1.1±0.12,28.67±6.07,17.4±2.37, the changes of PPARy expression are significant (P<0.01)among three stages. During pregnancy25d and40d in endometrial and chorionic, the expression levels of PPARy in embryonic survival group were higher than the embryo death group. Correlation analysis shows that the expression of PPARy in the endometrium and chorionic has no association with fetal weight、fetal length or sac length.Conclusion:During pregnancy25d,40d and70d in the chorion and endometrium of sow, the expression of PPARy shows a decreasing trend after the first rise. There is an correlation between the expression of PPARy in placenta and embryonic survival status, the expression levels of PPARy in embryonic survival group were higher than the embryo death group. The expression of PPARy in placenta had no association with fetal weight、fetal length or sac length.