Exposure to Progestin 17-OHPC Induces Gastrointestinal Dysfunction through Claudin-1 Suppression in Female Mice with Increased Anxiety-Like Behaviors

Abstract

Introduction

Progestin, commonly used in oral contraception and preventing preterm birth, elicits various off-target side effects on brain and gastrointestinal (GI) functions, yet the precise mechanisms remain elusive. This study aims to probe progestin’s impact on GI function and anxiety-like behaviors in female mice.

Methods

Colon stem cells were utilized to explore the mechanism underlying progestin 17-hydroxyprogesterone caproate (17-OHPC)-mediated suppression of claudin-1 (CLDN1), crucial for epithelial integrity. Chromatin immunoprecipitation and luciferase assays identified potential progestin-response elements on the CLDN1 promoter, with subsequent assessment of oxidative stress and pro-inflammatory cytokine release. Manipulation of vitamin D receptor (VDR) or estrogen receptor β (ERβ) expression elucidated their roles in 17-OHPC-mediated effects. Intestine-specific VDR deficient mice were generated to evaluate 17-OHPC’s impact on GI dysfunction and anxiety-like behaviors in female mice. Additionally, gene expression was analyzed in various brain regions, including the amygdala, hypothalamus, and hippocampus.

Results

Exposure to 17-OHPC suppressed CLDN1 expression via epigenetic modifications and VDR dissociation from the CLDN1 promoter. Furthermore, 17-OHPC intensified oxidative stress and pro-inflammatory cytokine release. VDR knockdown partly mimicked, while overexpression of either VDR or ERβ partly restored 17-OHPC-mediated effects. Intestinal VDR deficiency partly mirrored 17-OHPC-induced GI dysfunction, with minimal impact on 17-OHPC-mediated anxiety-like behaviors.

Conclusions

17-OHPC suppresses CLDN1 expression through VDR, contributing to GI dysfunction in female mice, distinct from 17-OHPC-induced anxiety-like behaviors. This study reveals a new mechanism and potential negative impact of progestin exposure on the GI tract, alongside inducing anxiety-like behaviors in female mice.

Plain Language Summary

Progestin is commonly used in oral contraceptives and for preventing preterm birth, but it exhibits various off-target side effects on brain and gastrointestinal (GI) function, the full extent of which remains largely unclear. In this study, we investigate the potential impact of progestin on the GI function in female mice with anxiety-like behaviors. Our findings reveal that exposure to the progestin 17-hydroxyprogesterone caproate (17-OHPC) suppresses the expression of claudin-1 (CLDN1) through epigenetic modifications and the dissociation of the vitamin D receptor (VDR) from the CLDN1 promoter. Additionally, 17-OHPC exposure exacerbates oxidative stress and the release of pro-inflammatory cytokines. Partial VDR deficiency in the intestine partly replicates the enhanced intestinal permeability and altered gut microbiota induced by 17-OHPC, though it has minimal effect on the anxiety-like behaviors triggered by 17-OHPC in female mice. In summary, progestin 17-OHPC suppresses CLDN1 expression via epigenetic alterations, contributing to GI dysfunction, distinct from progestin-induced anxiety-like behaviors. This sheds light on a novel mechanism and potential side effect of progestin exposure on GI system, alongside eliciting anxiety-like behaviors in female mice.