Marinobufagenin is an upstream modulator of Gadd45a stress signaling in preeclampsia

Preeclampsia (PE) is a hypertensive disorder of pregnancy, in which marinobufagenin (MBG), a circulating cardiotonic steroid, is increased. The Gadd45a stress sensor protein is an upstream modulator of the pathophysiological changes observed in PE. However, the effects of MBG on Gadd45a stress signaling remain unknown. We examined the expression of Gadd45a, the sFlt-1 receptor, and p38, as well as caspase 3 and 8 activities in placental samples from four groups of rats. These were: normal pregnant (NP, n = 8); pregnant rats which received weekly injections of desoxycorticosterone acetate and 0.9% saline as their drinking water (PDS, n = 9); normal pregnant rats injected with MBG (NPM, n = 8); and PDS rats injected with resibufogenin (RBG), an in vivo antagonist of MBG (PDSR, n = 8). Utilizing human cytotrophoblast (CTB) cells, we examined the effect of MBG on these stress signaling proteins in vitro. Placental Gadd45a expression, caspase 3 and 8 activities, sFlt-1 concentrations, and sFlt-1 receptor expression were significantly higher in PDS and NPM compared to NP and PDSR rats. Gadd45a protein was significantly upregulated in the CTB cells when MBG was present in concentrations ≥1 nM. Treatment with MBG (≥ 1 nM) also significantly arrested cell cycle progression and activated the expression of the Gadd45a-mediated stress signaling proteins. Inhibition of Gadd45a through RNAi-mediation attenuated MBG-induced CTB cell stress signaling. In conclusion, MBG is involved in the alteration in Gadd45a stress signaling both in vivo and in vitro and RBG prevents these changes when administered in vivo.

Research Highlights
► Placental Gadd45a expression was significantly higher in a rat model of preeclampsia.
► Caspase 3 and 8 activities and sFlt-1 receptor expression were higher in the rat model.
► MBG is involved in the alteration in Gadd45a stress signaling both in vivo and in vitro.
► RBG prevents these changes when administered in vivo.