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. 2025 Jan 12;14(2):214.
doi: 10.3390/foods14020214.

Combined BPA and DIBP Exposure Induced Intestinal Mucosal Barrier Impairment Through the Notch Pathway and Gut Microbiota Dysbiosis in Mice

Mengge Duan  1 , Yuting Wang  1 , Shiyu Chen  1 , Jiawen Lu  1 , Ruihong Dong  1 , Qiang Yu  1 , Jianhua Xie  1 , Yi Chen  1
Affiliations

Combined BPA and DIBP Exposure Induced Intestinal Mucosal Barrier Impairment Through the Notch Pathway and Gut Microbiota Dysbiosis in Mice

Mengge Duan et al. Foods. .

Abstract

Bisphenol A (BPA) and diisobutyl (DIBP) phthalate are widely used as typical plasticizers in food packaging. Plasticizers can be released from polymers, migrate into food, and be ingested by humans, leading to various health problems. However, little research has investigated the combined toxicity of BPA and DIBP, particularly their intestinal toxicity. Our goal is to analyse the combined toxicity of BPA (50 mg/kg) and DIBP (500 mg/kg) on the intestines of KM mice. Additionally, we tried to find natural products that can inhibit or prevent the combined toxicity of BPA and DIBP. The results indicated that the combination of BPA and DIBP exposure resulted in a reduction of beneficial flora, an increase in D-Lac levels (136 ± 14 μmol/L), an increase in intestinal permeability, activation of the notch pathway, and a decline in intestinal stem cells (ISCs) to goblet cells, compared to single-exposure sources. Nevertheless, Rubus chingii Hu phenolic extract (RHPE) (200, 400 and 600 mg/kg) ameliorated the BPA and DIBP-induced intestinal microbiota disruption and intestinal mucosal barrier impairment by inhibiting the overactivation of the notch pathway. The results of this study highlight the potential risks to human health posed by the combination of BPA and DIBP and may help explain the potential pathways of enterotoxicity caused by combined ingestion.

Keywords: BPA; DIBP; Rubus chingii Hu phenolic extract; gut microbiota; intestinal mucosal barrier; notch pathway.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Figure 1
Figure 1
Experimental grouping scheme (A) Schematic diagram of the animal experiment. (B) Cell grouping.
Figure 2
Figure 2
Effect of RHPE on organ index in mice. (A) Body weight change of the mice. (B) Liver index. Different letters indicated significant differences among groups (p < 0.05).
Figure 3
Figure 3
RHPE improved the pathological damage caused by BPA + DIBP combined exposure in mice. (A) Serum D-Lac levels in mice. (B) Representative images of H&E staining of ileum sections. (C,D) Villus height and crypt depth. (E,F) E-cad immunohistochemical staining analysis. Different letters indicated significant differences among groups (p < 0.05).
Figure 4
Figure 4
RHPE promotes the proliferation and differentiation of mouse intestinal stem cells. Immunohisto-chemical staining and relative positive area of (A,B) Ki67. (C,D) Lgr5. (E,F) Lyz. (G,H) Muc2. (I,J) AB–PAS staining of goblet cells. Different letters indicated significant differences among groups (p < 0.05).
Figure 4
Figure 4
RHPE promotes the proliferation and differentiation of mouse intestinal stem cells. Immunohisto-chemical staining and relative positive area of (A,B) Ki67. (C,D) Lgr5. (E,F) Lyz. (G,H) Muc2. (I,J) AB–PAS staining of goblet cells. Different letters indicated significant differences among groups (p < 0.05).
Figure 5
Figure 5
Caco-2/HT29-MTX Co-culture cytotoxicity of BPA, DIBP and RHPE. (A) BPA cell viability (24 h). (B) DIBP cell viability (24 h). (C) BPA + DIBP cell viability (24 h). (D) BPA and DIBP CI index analysis. (E) Effect of RHPE on cell viability. (F) BBR and NAC cell viability. Different letters indicated significant differences among groups (p < 0.05).
Figure 6
Figure 6
RHPE improved monolayer integrity and permeability of Caco-2/HT29-MTX co-cultured cells. (A) Monolayer integrity. (B) Permeability. Different letters indicated significant differences among groups (p < 0.05).
Figure 7
Figure 7
RHPE alleviates goblet cell damage through DLL4–Notch1–Hes1–Math1 pathway. (A,B) Relative protein expression of Muc2. (C) Notch pathway key protein expression was measured by Western blotting in Caco-2/HT29-MTX. (D–G) Notch1, DLL4, Hes1, Math1. Different letters indicated significant differences among groups (p < 0.05).
Figure 8
Figure 8
Effects of RHPE on gut microbiota in BPA- and DIBP-induced mice. (A) Chao1 index. (B) Observed-species index. (C) Simpson index. (D) Shannon index. (E,F) Principal coordinate analysis (PcoA) and NMDS of β diversity. Different letters indicated significant differences among groups (p < 0.05)
Figure 9
Figure 9
Effects of RHPE on intestinal microbial disorders in mice induced by combined exposure to BPA and DIBP. (A) Taxonomic composition at phylum level. (B) Taxonomic composition at genus level. (C) LDA analysis.
Figure 10
Figure 10
Effects of RHPE on the content of SCFAs in mice. (A) Relative content of SCFAs. (B) Acetic acid. (C) Propionic acid. (D) Butyric acid. Different letters indicated significant differences among groups (p < 0.05).
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