Abstract
Spontaneous preterm birth (sPTB) has been increasingly associated with alterations in vaginal microbiota. While Lactobacillus spp., which physiologically dominate the cervical microbiota, are considered protective with a lower risk of intra-amniotic infection and chorioamnionitis, other microbes such as Gardnerella vaginalis are associated with an increased risk of sPTB. Although this association is well documented, the host mechanisms that regulate the composition of vaginal microbial communities remain poorly understood. Emerging evidence suggests that host-derived exosomes may play a critical role in shaping the microbial environment. This study hypothesized that endothelial cell-derived exosomes may modulate the growth of probiotic Lactobacillus spp. through changes in microRNA cargo, thereby influencing the risk of sPTB. To investigate this hypothesis, HEC-1-A cells were stimulated with lipopolysaccharides (LPS), and exosomes were isolated from these cells. These exosomes were then applied to four Lactobacillus strains (L. crispatus, L. gasseri, L. jensenii, L. reuteri) to evaluate how LPS-induced alterations in exosomal microRNA content affect probiotic growth. The results indicate that exosomes derived from LPS-stimulated HEC-1-A cells inhibited the four bacterial strains and facilitated the expansion of the opportunistic pathogen G. vaginalis in a mixed-culture system. MicroRNA sequencing revealed that LPS stimulation increased the levels of miR-181d-5p and miR-181c in these exosomes, both of which may contribute to the suppression of Lactobacillus spp. growth. Taken together, these findings suggest a novel regulatory pathway in which host-derived exosomes influence the vaginal microbiota, suggesting that disruptions in this mechanism may contribute to vaginal dysbiosis and increase the risk of sPTB.
Data availability
All data generated or analysed during this study are included in this published article.
References
Ohuma, E. O. et al. National, regional, and global estimates of preterm birth in 2020, with trends from 2010: a systematic analysis. Lancet 402 (10409), 1261–1271. https://doi.org/10.1016/S0140-6736(23)00878-4 (2023).
Zivaljevic, J., Jovandaric, M. Z., Babic, S. & Raus, M. Complications of preterm birth-the importance of care for the outcome: A narrative review. Med. (Kaunas) 60 (6). https://doi.org/10.3390/medicina60061014 (2024).
Perin, J. et al. Global, regional, and national causes of under-5 mortality in 2000-19: An updated systematic analysis with implications for the sustainable development goals. Lancet Child. Adolesc. Health. 6 (2), 106–115. https://doi.org/10.1016/S2352-4642(21)00311-4 (2022).
Green, E. S. & Arck, P. C. Pathogenesis of preterm birth: bidirectional inflammation in mother and fetus. Semin Immunopathol. 42 (4), 413–429. https://doi.org/10.1007/s00281-020-00807-y (2020).
Daskalakis, G. et al. Maternal infection and preterm birth: From molecular basis to clinical implications. Child. (Basel) 10 (5). https://doi.org/10.3390/children10050907 (2023).
Migale, R. et al. Specific lipopolysaccharide serotypes induce differential maternal and neonatal inflammatory responses in a murine model of preterm labor. Am. J. Pathol. 185 (9), 2390–2401. https://doi.org/10.1016/j.ajpath.2015.05.015 (2015).
Bayar, E., Bennett, P. R., Chan, D., Sykes, L. & MacIntyre, D. A. The pregnancy microbiome and preterm birth. Semin Immunopathol. 42 (4), 487–499. https://doi.org/10.1007/s00281-020-00817-w (2020).
Kindinger, L. M. et al. The interaction between vaginal microbiota, cervical length, and vaginal progesterone treatment for preterm birth risk. Microbiome 5 (1), 6. https://doi.org/10.1186/s40168-016-0223-9 (2017).
Callahan, B. J. et al. Replication and refinement of a vaginal microbial signature of preterm birth in two racially distinct cohorts of US women. Proc. Natl. Acad. Sci. U S A. 114 (37), 9966–9971. https://doi.org/10.1073/pnas.1705899114 (2017).
DiGiulio, D. B. et al. Temporal and spatial variation of the human microbiota during pregnancy. Proc. Natl. Acad. Sci. U S A. 112 (35), 11060–11065. https://doi.org/10.1073/pnas.1502875112 (2015).
Fettweis, J. M. et al. The vaginal microbiome and preterm birth. Nat. Med. 25 (6), 1012–1021. https://doi.org/10.1038/s41591-019-0450-2 (2019).
Werter, D. E. et al. The Risk of Preterm Birth in Low Risk Pregnant Women with Urinary Tract Infections. Am. J. Perinatol. 40 (14), 1558–1566. https://doi.org/10.1055/s-0041-1739289 (2023).
Wang, E., Tang, P. & Chen, C. Urinary tract infections and risk of preterm birth: a systematic review and meta-analysis. Rev. Inst. Med. Trop. Sao Paulo. 66, e54. https://doi.org/10.1590/S1678-9946202466054 (2024).
Mohanty, T., Doke, P. P. & Khuroo, S. R. Effect of bacterial vaginosis on preterm birth: a meta-analysis. Arch. Gynecol. Obstet. 308 (4), 1247–1255. https://doi.org/10.1007/s00404-022-06817-5 (2023).
Kalluri, R. & LeBleu, V. S. The biology, function, and biomedical applications of exosomes. Science 367 (6478). https://doi.org/10.1126/science.aau6977 (2020).
Sultan, S., Mottawea, W., Yeo, J. & Hammami, R. Gut microbiota extracellular vesicles as signaling molecules mediating host-microbiota communications. Int. J. Mol. Sci. 22 (23). https://doi.org/10.3390/ijms222313166 (2021).
Song, Y., Shi, M. & Wang, Y. Deciphering the role of host-gut microbiota crosstalk via diverse sources of extracellular vesicles in colorectal cancer. Mol. Med. 30 (1), 200. https://doi.org/10.1186/s10020-024-00976-8 (2024).
Foster, B. P. et al. Extracellular vesicles in blood, milk and body fluids of the female and male urogenital tract and with special regard to reproduction. Crit. Rev. Clin. Lab. Sci. 53 (6), 379–395. https://doi.org/10.1080/10408363.2016.1190682 (2016).
Wang, L-M., Lee, B-H., Hou, C-Y., Hsu, W-H. & Tai, C-J. Probiotics-derived extracellular vesicles protect oxidative stress against h2o2 induction in placental cells. Fermentation 8 (2). https://doi.org/10.3390/fermentation8020074 (2022).
Lee, B. H. et al. The applications of Lactobacillus plantarum-derived extracellular vesicles as a novel natural antibacterial agent for improving quality and safety in tuna fish. Food Chem. 340, 128104. https://doi.org/10.1016/j.foodchem.2020.128104 (2021).
Lin, L. T., Shi, Y. C., Choong, C. Y. & Tai, C. J. The fruits of paris polyphylla inhibit colorectal cancer cell migration induced by fusobacterium nucleatum-derived extracellular vesicles. Molecules 26 (13). https://doi.org/10.3390/molecules26134081 (2021).
Chin, W. L. et al. Small intestine-residing probiotics suppress neurotoxic bile acid production via extracellular vesicle-mediated inhibition of Clostridium scindens. Food Res. Int. 207, 116049. https://doi.org/10.1016/j.foodres.2025.116049 (2025).
Rueda-Robles, A., Rodriguez-Lara, A., Meyers, M. S., Saez-Lara, M. J. & Alvarez-Mercado, A. I. Effect of probiotics on host-microbiota in bacterial infections. Pathogens 11 (9). https://doi.org/10.3390/pathogens11090986 (2022).
Smythies, L. E. & Smythies, J. R. Exosomes in the gut. Front. Immunol. 5, 104. https://doi.org/10.3389/fimmu.2014.00104 (2014).
Li, L., Kang, J. & Lei, W. Role of Toll-like receptor 4 in inflammation-induced preterm delivery. Mol. Hum. Reprod. 16 (4), 267–272. https://doi.org/10.1093/molehr/gap106 (2010).
Edey, L. F. et al. The local and systemic immune response to intrauterine LPS in the prepartum mouse. Biol. Reprod. 95 (6), 125. https://doi.org/10.1095/biolreprod.116.143289 (2016).
Okawa, T. et al. Effect of lipopolysaccharide on uterine contractions and prostaglandin production in pregnant rats. Am. J. Obstet. Gynecol. 184 (2), 84–89. https://doi.org/10.1067/mob.2001.108083 (2001).
Torbe, A. et al. Maternal plasma lipopolysaccharide binding protein (LBP) concentrations in pregnancy complicated by preterm premature rupture of membranes. Eur. J. Obstet. Gynecol. Reprod. Biol. 156 (2), 153–157. https://doi.org/10.1016/j.ejogrb.2011.01.024 (2011).
Joo, E. et al. Maternal plasma and amniotic pentraxin 3, resistin, and IGFBP-3: biomarkers of microbial invasion of amniotic cavity and/or intra-amniotic inflammation in women with preterm premature rupture of membranes. J. Korean Med. Sci. 36 (44), e279. https://doi.org/10.3346/jkms.2021.36.e279 (2021).
Wang, J. et al. Translocation of vaginal microbiota is involved in impairment and protection of uterine health. Nat. Commun. 12 (1), 4191. https://doi.org/10.1038/s41467-021-24516-8 (2021).
Ciesielska, A., Matyjek, M. & Kwiatkowska, K. TLR4 and CD14 trafficking and its influence on LPS-induced pro-inflammatory signaling. Cell. Mol. Life Sci. 78 (4), 1233–1261. https://doi.org/10.1007/s00018-020-03656-y (2021).
Bell-Hensley, A., Das, S. & McAlinden, A. The miR-181 family: Wide-ranging pathophysiological effects on cell fate and function. J. Cell. Physiol. 238 (4), 698–713. https://doi.org/10.1002/jcp.30969 (2023).
Rezaei, T. et al. microRNA-181 serves as a dual-role regulator in the development of human cancers. Free Radic. Biol. Med. 152, 432–454. https://doi.org/10.1016/j.freeradbiomed.2019.12.043 (2020).
Maggie R., Williams Robert D., Stedtfeld James M., Tiedje Syed A., Hashsham (2017) MicroRNAs-Based Inter-Domain Communication between the Host and Members of the Gut Microbiome Frontiers in Microbiology 810.3389/fmicb.2017.01896
Ce, Yuan Michael B., Burns Subbaya, Subramanian Ran, Blekhman Interaction between Host MicroRNAs and the Gut Microbiota in Colorectal Cancer ABSTRACT mSystems 3(3), https://doi.org/10.1128/mSystems.00205-17 (2018).
Funding
This study was supported by the Taipei Municipal Wan Fang Hospital, Taiwan, R.O.C. (112-wf-phd-01). LTL is supported by the National Science and Technology Council (NSTC) of Taiwan (NSTC 114-2320-B-038-044).
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LMW and YCS conceived the study and performed the formal analysis. BHL contributed to the methodology, investigation, and visualization, with additional methodological input from EWL. CJT and LTL supervised the work. YCS, CHL, and LTL contributed to the writing. All authors reviewed the manuscript.
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YCS is employed as a Research and Development Manager at BioX Biotech CO., Ltd., which supplied materials used in this study. The company had no role in the design, data collection, analysis, interpretation, or writing of the manuscript. The authors declare no other competing interests.
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Wang, LM., Shi, YC., Lee, BH. et al. LPS-induced endometrial cell-derived exosomes suppress probiotic Lactobacillus growth. Sci Rep (2026). https://doi.org/10.1038/s41598-026-44830-9
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DOI: https://doi.org/10.1038/s41598-026-44830-9
