Bone Marrow Mesenchymal Stem Cells Attenuate Mitochondria Damage Induced by Hypoxia in Mouse Trophoblasts

Abstract

ObjectiveWe aimed to observe the change of mitochondrial function and structure as well as the cell function induced by hypoxia in mouse trophoblasts, and moreover, to validate the restoration of these changes after co-culture with bone marrow mesenchymal stem cells (hereinafter referred to as “MSCs”). Further, we explored the mechanism of MSCs attenuating the functional damage of trophoblasts caused by hypoxia.MethodsCells were divided into two groups, trophoblasts and MSCs+trophoblasts respectively, and the two groups of cells were incubated with normoxia or hypoxia. Chemiluminescence was used to assay the β-HCG and progesterone in cell culture supernatants quantitatively. Western blotting and PCR were applied to detect the expression of Mfn2, MMP-2, MMP-9 and integrin β1 in the two groups. The mitochondrial membrane potential of each group of cells was detected with JC-1 dye and the ATP content was measured by the phosphomolybdic acid colorimetric method. We utilized transmission electron microscopy for observing the ultrastructure of mitochondria in trophoblasts. Finally, we assessed the cell apoptosis with flow cytometry (FCM) and analyzed the expression of the apoptosis related genes—Bcl-2, Bax, Caspase3 and Caspase9 by western blotting.ResultsThe results showed that the Mfn2 expression was reduced after 4 h in hypoxia compared with that in normoxia, but increased in the co-culture group when compared with that in the separated-culture group (p<0.05). In addition, compared with the separated-culture group, theβ-HCG and progesterone levels in the co-culture group were significantly enhanced (p<0.05), and so were the expressions of MMP-2, MMP-9 and integrin β1 (p<0.05). Moreover, it exhibited significantly higher in ATP levels and intensified about the mitochondrial membrane potential in the co-culture group. TEM revealed disorders of the mitochondrial cristae and presented short rod-like structure and spheroids in hypoxia, however, in the co-culture group, the mitochondrial cristae had a relatively regular arrangement and the mitochondrial ultrastructure showed hyperfusion. The expression of Bax, Caspase3 and Caspase9 was decreased in the co-culture group when compared with that in trophoblast cells cultured alone (p<0.05), while the Bcl-2 levels and the Apoptosis Index (AI) were markedly increased in the co-culture group (p<0.05).ConclusionBone marrow mesenchymal stem cells can attenuate mitochondria damage and cell apoptosis induced by hypoxia; the mechanism could be upregulating the expression of Mfn2 in mouse trophoblasts and changing mitochondrial structure.

Diagnostic biomolecules and combination therapy for pre-eclampsia

“…In this case, MSCs can significantly improve the symptoms of Th1 cells of PE mouse model, implying that MSCs may play a significant role in maternal interface immunity [ 158 ]. Consistent with these, another study demonstrated that MSCs could improve the functions of mouse trophoblasts and endothelial cells to promote angiogenesis through attenuating the hypoxia-induced mitochondria damage [ 159 ]. As for the underlying mechanisms, there is increasing evidence that the beneficial effects of MSCs are exerted through the secretory bodies and reducing SFLT-1 to restore angiogenesis.…”

Phenylbutyric acid inhibits hypoxia‐induced trophoblast apoptosis and autophagy in preeclampsia via the PERK/ATF‐4/CHOP pathway

“…Apoptosis is frequently accompanied by abnormal mitochondrial morphology and function (Lv et al, 2021; Wang et al, 2022). It has been found that hypoxia triggers mitochondrial damage in mouse trophoblast cells (Wang et al, 2016). Inhibition of the mitochondrial apoptotic pathway attenuates hypoxia‐induced trophoblast cell damage (Ma et al, 2020).…”

The pathological and therapeutic roles of mesenchymal stem cells in preeclampsia

“…Hypoxia brings on disarrangement of mitochondrial ultrastructure in mice trophoblasts in vitro. bm-MSCs intensify mitochondrial membrane potential and increase ATP production/consumption in trophoblast and support cell survival against hypoxia (100). PD−MSCs reduce 10.3389/fmed.2022.923334 mitochondrial damage by downregulating Heat shock protein 60 (HSP60) (inducted under mitochondrial stress) and upregulating prohibitin 1 (PHB1) (involved in stabilizing mitochondria) expression and promoting ATP generation/consumption by upregulating VDAC in the mitochondria of trophoblast cells (101).…”