Introduction Immune thrombocytopenia (ITP) affects approximately 1 to10 per 10,000 pregnancies. It showed that bone marrow mesenchymal stem cells (BM-MSCs) in ITP patients during pregnancy exhibit significant impairment, including increased senescence, apoptosis, and compromised immunoregulatory functions as our previous study. Progesterone has been identified as a critical metabolite influencing the composition of gut microbiota, which in turn contributes to BM-MSCs injury in ITP during pregnancy (J Thromb Haemost,2025). Nevertheless, the precise mechanism by which progesterone-mediated alterations in gut microbiota lead to MSCs injury in ITP during pregnancy remains inadequately understood. It is also needed to explore the potential of targeting gut microbiota as a therapeutic strategy for managing pregnant ITP patients.

Methods Fresh fecal samples were collected from pregnant ITP patients, non-pregnant ITP patients,and healthy controls. Humanized fecal microbiota mice were development through fecal microbiota transplantation (FMT). The gut microbiota profiles of mice were analyzed using 16S rDNA sequencing technology. Subsequently, an ITP model was established. The senescence and apoptosis of BM-MSCs in mice with different fecal microbiota sources were assessed, and these MSCs were co-cultured with CD34+cells to examine their effects on megakaryocytes (MKs) maturation and platelet production. Serum levels of NOD1 ligand (NOD1L) in FMT mice were measured. C12-iE-DAP was used to treat MSCs in vitro to investigate changes in MSCs senescence, apoptosis, and the expression of RIP2, IRF3, JAK1 and STAT1 before and after C12-iE-DAP intervention. The levels of cytokines secreted by MSCs were also measured. In vivo, mice of different groups were administered probiotics or C12-iE-DAP via gavage. Platelet counts and serum NOD1L levels in the mice were subsequently determined.

Results Fresh fecal samples were collected from three groups: ITP patients during pregnancy, non-pregnant ITP patients, and healthy controls. Mice were then inoculated with fecal microbiota from these resources, resulting in three experimental groups: the ITP during pregnancy-FMT, the ITP-FMT and the HC-FMT group. The gut microbiota profiles analysis indicated that the α-diversity of the gut microbiota in the ITP during pregnancy-FMT group was lower compared to both the ITP-FMT and HC-FMT groups. Furthermore, the abundance of Gram-negative bacteria was significantly reduced in the ITP during pregnancy-FMT group. The abundances of unclassified Ruminococcaceae, Intestinibacter, Barnesiella, Staphylococcus, and Bacillus were increased in this group.

BM-MSCs were subsequently isolated from the recipient mice, revealing that the proportion of senescent and apoptotic cells was significantly higher in ITP during pregnancy-FMT-MSCs compared to other groups. The MSCs from different groups were co-cultured with CD34+ cells, which were induced to differentiate into MKs. In the group with ITP during pregnancy-FMT-MSCs, there was a significant reduction in the proportion of CD41+CD42b+ MKs and platelets. Concurrently, serum levels of NOD1L were significantly lower in ITP during pregnancy-FMT recipients. Subsequently, MSCs from ITP during pregnancy-FMT mice were stimulated in vitro with C12-iE-DAP. Following stimulation with C12-iE-DAP, there was a notable decrease in the proportion of senescent and apoptotic MSCs. Additionally, there was an upregulation in the expression of RIP2, IRF3, JAK1, STAT1, and an increase in the level of IFN-I in the supernatant. The proportion of CD41+CD42b+ MKs and platelets also significantly increased when co-cultured with bone marrow-derived CD34+ cells.

Finally, established mouse models of ITP during pregnancy mice and ITP during pregnancy-FMT mice were randomly assigned to control and experimental groups. The experimental groups received gavage administration of probiotics and C12-iE-DAP, respectively. Elevated levels of NOD1L and platelet counts were observed in the experimental groups.

Conclusions Supplementation with probiotics, elevation of NOD1L levels, and activation of the NOD1 receptor signaling pathway, which subsequently triggers the IFN-I/STAT1 signaling cascade, may ameliorate the senescence and apoptosis of MSCs in ITP during pregnancy, thereby promoting platelet production. This approach represents a potentially novel therapeutic strategy for managing ITP during pregnancy.

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2025
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