FPR1 Antagonist (BOC-MLF) Inhibits Amniotic Epithelial-mesenchymal Transition

Abstract

Objective

Premature rupture of membranes (PROM) is a common pregnancy disorder that is closely associated with structural weakening of fetal membranes. Studies have found that formyl peptide receptor 1 (FPR1) activates inflammatory pathways and amniotic epithelialmesenchymal transition (EMT), stimulates collagen degradation, and leads to membrane weakening and membrane rupture. The purpose of this study was to investigate the anti-inflammatory and EMT inhibitory effects of FPR1 antagonist (BOC-MLF) to provide a basis for clinical prevention of PROM.

Methods

The relationship between PROM, FPR1, and EMT was analyzed in human fetal membrane tissue and plasma samples using Western blotting, PCR, Masson staining, and ELISA assays. Lipopolysaccharide (LPS) was used to establish a fetal membrane inflammation model in pregnant rats, and BOC-MLF was used to treat the LPS rat model. We detected interleukin (IL)-6 in blood from the rat hearts to determine whether the inflammatory model was successful and whether the anti-inflammatory treatment was effective. We used electron microscopy to analyze the structure and collagen expression of rat fetal membrane.

Results

Western blotting, PCR and Masson staining indicated that the expression of FPR1 was significantly increased, the expression of collagen was decreased, and EMT appeared in PROM. The rat model indicated that LPS caused the collapse of fetal membrane epithelial cells, increased intercellular gaps, and decreased collagen. BOC-MLF promoted an increase in fetal membrane collagen, inhibited EMT, and reduced the weakening of fetal membranes.

Conclusion

The expression of FPR1 in the fetal membrane of PROM was significantly increased, and EMT of the amniotic membrane was obvious. BOC-MLF can treat inflammation and inhibit amniotic EMT.

Keywords

formyl peptide receptor 1 / BOC-MLF / epithelial-mesenchymal transition / premature rupture of membranes

References

[1] SchmitzT, SentilhesL, LortheE, et al.. Preterm premature rupture of the membranes: Guidelines for clinical practice from the French College of Gynaecologists and Obstetricians (CNGOF). Eur J Obstet Gynecol Reprod Biol, 2019, 236: 1-6

[2] Practice Bulletin No. 172: Premature Rupture of Membranes. Obstet Gynecol, 2016,128(4):e165–e177

[3] PasquierJC, DoretM. Fetal membranes: embryological development, structure and the physiopathology of the preterm premature rupture of membranes. J Gynecol Obstet Biol Reprod (Paris), 2008, 37(6): 579-588

[4] StrevensH, AllenK, ThorntonJG. Management of premature prelabor rupture of the membranes. Ann N Y Acad Sci, 2010, 1205: 123-129

[5] LannonSM, VanderhoevenJP, EschenbachDA, et al.. Synergy and interactions among biological pathways leading to preterm premature rupture of membranes. Reprod Sci, 2014, 21(10): 1215-1227

[6] MogamiH, WordRA. Healing Mechanism of Ruptured Fetal Membrane. Front Physiol, 2020, 11: 623

[7] PreveteN, LiottiF, ViscianoC, et al.. The formyl peptide receptor 1 exerts a tumor suppressor function in human gastric cancer by inhibiting angiogenesis. Oncogene, 2015, 34(29): 3826-3838

[8] Mogami H. Mini-review: Wound healing of amnion and macrophages. J Obstet Gynaecol Res, 48(3):563–567

[9] LeY, YangY, CuiY, et al.. Receptors for chemotactic formyl peptides as pharmacological targets. Int Immunopharmacol, 2002, 2(1): 1-13

[10] GaoL, ZengN, YuanZ, et al.. Knockout of Formyl Peptide Receptor-1 Attenuates Cigarette Smoke-Induced Airway Inflammation in Mice. Front Pharmacol, 2021, 12: 632225

[11] ChenH, ZhangL. Downregulation of FPR1 abates lipopolysaccharide-induced inflammatory injury and apoptosis by upregulating MAPK signaling pathway in murine chondrogenic ATDC5 cells. Allergol Immunopathol (Madr), 2021, 49(5): 57-63

[12] TchirikovM, Schlabritz-LoutsevitchN, MaherJ, et al.. Mid-trimester preterm premature rupture of membranes (PPROM): etiology, diagnosis, classification, international recommendations of treatment options and outcome. J Perinat Med, 2018, 46(5): 465-488

[13] HudallaH, KarenbergK, KuonRJ, et al.. LPS-induced maternal inflammation promotes fetal leukocyte recruitment and prenatal organ infiltration in mice. Pediatr Res, 2018, 84(5): 757-764

[14] ViziES, SzelényiJ, SelmeczyZS, et al.. Enhanced tumor necrosis factor-alpha-specific and decreased interleukin-10-specific immune responses to LPS during the third trimester of pregnancy in mice. J Endocrinol, 2001, 171(2): 355-361

[15] JanzenC, SenS, LeiMY, et al.. The Role of Epithelial to Mesenchymal Transition in Human Amniotic Membrane Rupture. J Clin Endocrinol Metab, 2017, 102(4): 1261-1269

[16] VanderhoevenJP, BierleCJ, KapurRP, et al.. Group B streptococcal infection of the choriodecidua induces dysfunction of the cytokeratin network in amniotic epithelium: a pathway to membrane weakening. PLoS Pathog, 2014, 10(3): e1003920

[17] MenonR, FortunatoSJ. The role of matrix degrading enzymes and apoptosis in rupture of membranes. J Soc Gynecol Investig, 2004, 11(7): 427-437

[18] Vadillo-OrtegaF, SadowskyDW, HaluskaGJ, et al.. Identification of matrix metalloproteinase-9 in amniotic fluid and amniochorion in spontaneous labor and after experimental intrauterine infection or interleukin-1 beta infusion in pregnant rhesus monkeys. Am J Obstet Gynecol, 2002, 186(1): 128-138

[19] ChattopadhyayI, AmbatiR, GundamarajuR. Exploring the Crosstalk between Inflammation and Epithelial-Mesenchymal Transition in Cancer. Mediators Inflamm, 2021, 2021: 9918379

[20] ZhangQ, RaoofM, ChenY, et al.. Circulating mitochondrial DAMPs cause inflammatory responses to injury. Nature, 2010, 464(7285): 104-107

[21] RabietMJ, HuetE, BoulayF. The N-formyl peptide receptors and the anaphylatoxin C5a receptors: an overview. Biochimie, 2007, 89(9): 1089-1106

[22] ChiangSF, HuangKC, ChenWT, et al.. Polymorphism of formyl peptide receptor 1 (FPR1) reduces the therapeutic efficiency and antitumor immunity after neoadjuvant chemoradiotherapy (CCRT) treatment in locally advanced rectal cancer. Cancer Immunol Immunother, 2021, 70(10): 2937-2950

[23] YangSC, ChangSH, HsiehPW, et al.. Dipeptide HCH6-1 inhibits neutrophil activation and protects against acute lung injury by blocking FPR1. Free Radic Biol Med, 2017, 106: 254-269