Differential Expression Of Galectin-3 In Fetal Rat Penis Following Maternal Exposure To Di(nbutyl) Phthalate

Objectives: To isolate and identify differentially expression proteins in fetal rat penis following maternal exposure to di(nbutyl)phthalate (DBP), and to verify the differential expression of galectin-3 in order to further explore the mechanism of hypospadias.Methods: Twenty pregnant rats were randomly divided into two groups and were daily treated by gavage with 800mg/kg DBP or 2ml soybean oil from the 14th day to the 18th day of pregnancy. The penis of normal and hypospadiac male fetal rats were respectively taken out on gestation day 19, and total penis proteins were extracted. The proteins were separated by two-dimensional gel electrophoresis and identified by mass spectrometry, and then to verify the differential expression of galectin-3 by Western blotting and immunohistochemistry.Results: The incidence of hypospadias was 40%.Fifty-one differentially expressed protein spots have been found and 21 of which were identified by mass spectrometry and searched in the SwissProt database, such as galectin-3, Lamin B1, LDH-B, etc. These proteins are mainly related to tissue differentiation and growth regulation and so on. Using Western blotting, we found that the expression of galectin-3 in the experimental group was significantly lower than the control group, by immunohistochemistry and found that galectin-3 was mainly localized in the epithelial cells of the urethra, the control group staining intensity was stronger than the experimental group.Conclusions: Proteomics is an effective method for studying the reproductive and developmental toxicity of DBP, some of these differentially expressed proteins may provide evidence for us to further investigate the molecular mechanism of hypospadias induced by DBP. DBP can reduce the galectin-3 expression in epithelium, affect cellular proliferation, apoptosis and urethral fold fusion, then result in the occurrence of hypospadias. But its function needs further study, other proteins in this experiment also needs to be validated and in-depth study.