Combining experiments and bioinformatics to identify transforming growth factor-β1 as a key regulator in angiotensin II-induced trophoblast senescence.

INTRODUCTION: Accelerated senescence of trophoblast may cause several diverse pregnancy outcomes; however, the cause of accelerated trophoblast senescence remains unclear. The renin-angiotensin system (RAS) is closely related to organ senescence. Therefore, in the present study, we hypothesized that angiotensin (Ang)II, one of the most important RAS family members, accelerates trophoblast senescence through the transforming growth factor β-1 (TGF-β1) pathway.

METHODS: AngII and Ang1-7 were used to stimulate pregnant rats. AngII and its inhibitor olmesartan were used to stimulate trophoblast. Thereafter, senescence levels were measured. Furthermore, we used AngII to stimulate trophoblast and utilized RNA-sequencing (RNAseq) to analyze the expression of differentially expressed genes (DEGs). After identifying the overlapping genes by comparing the DEGs and senescence-related genes, we employed CytoHubba software to calculate the top five hub genes and selected TGF-β1 as the target gene. We transfected the AngII-stimulated trophoblast with TGF-β1 small interfering RNA (siRNA) and measured the senescence levels.

RESULTS: Senescence markers were upregulated in the AngII group compared with that in the control group. Furthermore, following AngII stimulation and RNAseq measurement, we identified 607 DEGs and 13 overlapping genes. The top five hub genes were as follows: PLAU, PTGS2, PDGF-β, TGF-β1, and FOXO3. Upon knockdown of TGF-β1 expression in AngII-stimulated trophoblast using TGF-β1 siRNA, we observed a downregulation of p53 and p62 mRNA expression.

DISCUSSION: AngII accelerates trophoblast senescence through the TGF-β1 pathway.