Role of hydrogen sulphide in regulating invasion in trophoblast cells

Abstract

IntroductionHydrogen sulphide is recently being considered as endogenous gaseous mediator which has shown to be involved in regulating many physiological processes like cell proliferation migration, invasion and phenomena like oxidative stress, angiogenesis, vasoregulation and neuromodulation. However, its role in modulating invasion in trophoblast cells, the key component of placenta is not known. Hence, in the present study we sought to explore the effects of hydrogen sulphide (H2S) on invasion capacity of trophoblast cells using HTR‐8/SVneo cell lineMaterial and methodsThe invasion potential of HTR‐8/SVneo cells was assessed by transwell invasion assay. The Matrigel invasion assay was performed at 37°C using 24 well transwell inserts (corning Greiner Bio‐One) coated with semi solid Matrigel 10% (Becton and Dickinson). The HTR‐8/SVneo cells received various treatments and then were transferred to the transwell(Boyden's) chamber. The cells were exposed to NaHS (sodium hydrosulphide:Hydrogen sulphide chemical mimic), its inhibitor AOAA (amino‐oxiacetate) in different doses for various time periods and hypoxia /reoxygenation. Cells (2500) suspended in 200ul of serum free medium were seeded onto the upper chamber. However,400ul of 10% FBS was added to lower chamber. Matrigel was removed from upper surface. Cells that migrated and invaded through the membrane to the under surface were methanol fixed and stained by 0.5% crystal violet. The cells were counted under Nikon Ti‐S Eclipse microscope using NIS‐AR software. The percentage invasion was calculated by the formula: mean number of cells invading the Matrigel membrane (test group) / mean number of cells invading through the insert membrane (control group) X100ResultsThe Percentage invasion was much higher when the cells received hydrogen sulphide mimic i.e. NaHS 100mM for 10 minutes (220%) as compared to cells which did not receive any treatment (100%). The trophoblast cells which were subjected to hypoxia reoxygenation (two cycles) showed reduction in the percentage invasion (20%). However, the cells of hypoxia reoxygenation treatment group regained their invasion capacity (46%) after they were exposed to NaHS 100 μM. On the other hand, the treatment of cells using AOAA [Cystathionine beta synthase (H2S producing enzyme) inhibitor] treatment decreased the cell invasion capacity (32%)ConclusionThe results from present study point towards the role of H2S in invasion of trophoblast cells, thereby predicting its role if any in placentation. These findings may also indicate that decrease in H2S may contribute to the pathogenesis of preeclampsia and also the other clinical outcomes associated with it like Fetal growth restriction (FGR)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.