Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Jun 9:12:678772.
doi: 10.3389/fphar.2021.678772. eCollection 2021.

Chlorogenic Acid Ameliorates Damage Induced by Fluorene-9-Bisphenol in Porcine Sertoli Cells

Shaoxuan Zhang  1 , Boxing Sun  1 , Dali Wang  1 , Ying Liu  1 , Jing Li  1 , Jiajia Qi  1 , Yonghong Zhang  1 , Chunyan Bai  1 , Shuang Liang  1
Affiliations

Chlorogenic Acid Ameliorates Damage Induced by Fluorene-9-Bisphenol in Porcine Sertoli Cells

Shaoxuan Zhang et al. Front Pharmacol. .

Abstract

4,4'-(9-Fluorenylidene) diphenol (BPFL, also known as BHPF and fluorene-9-bisphenol) is a novel bisphenol A substitute that is used in the plastics industry as an organic synthesis intermediate and is a potential endocrine disruptor. However, the deleterious effects of BPFL on porcine Sertoli cells (SCs) and the possible underlying mechanisms are still unclear. Chlorogenic acid (CA) is a free radical scavenger in the cellular antioxidant system that prevents oxidative damage and apoptosis. In the present research, we found that BPFL induced impairments in porcine SCs in a dose-dependent manner and that CA protected porcine SCs against BPFL exposure-induced impairments. Cell viability, proliferation and apoptosis assay results revealed that BPFL exposure could inhibit porcine SC proliferation and induce apoptosis, while CA supplementation ameliorated the effects of BPFL. Further analysis revealed that BPFL exposure induced oxidative stress, mitochondrial membrane potential dysfunction and DNA damage accumulation. Transcriptome analysis and further real-time quantitative PCR and Western blot results showed that BPFL exposure induced endoplasmic reticulum stress and apoptosis. Supplementation with CA dramatically ameliorated these phenotypes in BPFL-exposed porcine SCs. Overall, the present research reveals the possible underlying mechanisms by which BPFL exposure induced impairments and CA supplementation protected against these impairments in porcine SCs.

Keywords: BPFL; ameliorates; chlorogenic acid; impairments; porcine sertoli cells.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Chlorogenic acid (CA) ameliorates the BPFL exposure-induced decrease in cell viability in porcine Sertoli cells (SCs). (A, B) Porcine SCs were exposed to 0–100 μM for 24 and 48 h at 37°C, and cell viability was detected by CCK-8 assay respectively. (C) The viability of porcine SCs after 48 h of treatment with BPFL (50 μM) with/without CA (0–200 μM). (**p < 0.01) indicates a significant difference from the control (0.1% DMSO).
FIGURE 2
FIGURE 2
CA ameliorates BPFL exposure-induced proliferation arrest and apoptosis in porcine SCs. (A) Micrographs showing the incorporation of EDU in porcine SCs treated with BPFL and CA after 48 h. The proliferating cells fluoresce in red. The nuclei (Hoechst) are shown in blue. (B) Quantitative analysis of proliferation level was calculated by the percentage of red cells versus blue cells. (C) Cell apoptosis was detected by flow cytometry with Annexin V-FITC/PI double staining. (D) Analysis of the percentage of apoptotic cells after different treatments is shown in a histogram. (**p < 0.01) indicates a significant difference from the control (0.1% DMSO).
FIGURE 3
FIGURE 3
RNA-seq analysis of the effects of BPFL exposure on SCs. (A) The number of genes differentially expressed after BPFL treatment. (B) Volcano plot analysis of genes differentially expressed after BPFL treatment. (C) Gene Ontology enrichment analysis of the differential gene expression after BPFL treatment. (D) Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of genes downregulated after BPFL treatment. (E) KEGG enrichment analysis of genes upregulated after BPFL treatment.
FIGURE 4
FIGURE 4
CA ameliorates BPFL exposure-induced oxidative stress and DNA damage in porcine SCs. (A) Fluorescence photomicrographs of reactive oxygen species in cultured SCs after BPFL and CA treatment for 48 h. (B) Analysis of the relative reactive oxygen species levels in different treatment groups. (C) SOD activity of SCs after treatment. (D) Representative images of DNA damage in porcine SCs after single-cell gel electrophoresis assay. (E, F) The tail moment values and the tail DNA% of the comet tails were analyzed by ImageJ. (*p < 0.05) indicates a difference from the control (0.1% DMSO); (**p < 0.01) indicates a significant difference from the control (0.1% DMSO).
FIGURE 5
FIGURE 5
CA ameliorates BPFL exposure-induced mitochondrial dysfunction, ERS, apoptosis and autophagy in porcine SCs. (A) Fluorescence photomicrographs of JC-1 aggregates (green) and JC-1 monomers (red) of cultured SCs after BPFL and CA treatment for 48 h. (B) Analysis of the proportion of mitochondrial depolarization in different treatment groups. (C) Western blotting was used to detect the protein expression levels in each treatment group. (D) Histogram showing the relative expression of several proteins in each group compared to the reference proteins in the control group. (E) Relative mRNA expression of genes in different treatment groups. ns indicates no difference from the control (0.1% DMSO); (*p < 0.05) indicates a difference from the control; and (**p < 0.01) indicates a significant difference from the control.
FIGURE 6
FIGURE 6
Schematic diagram illustrating the effect of CA in ameliorating BPFL exposure-induced impairments in porcine SCs. BPFL exposure impaired porcine SCs by inducing oxidative stress, mitochondrial function, DNA damage, ERS and apoptosis. CA supplementation ameliorated the impairments induced by BPFL exposure in porcine SCs.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Alfhili M. A., Hussein H. A. M., Park Y., Lee M. H., Akula S. M. (2021). Triclosan Induces Apoptosis in Burkitt Lymphoma-Derived BJAB Cells through Caspase and JNK/MAPK Pathways. Apoptosis 26, 96–110. 10.1007/s10495-020-01650-0 - DOI - PubMed
    1. Alves M. G., Martins A. D., Vaz C. V., Correia S., Moreira P. I., Oliveira P. F., et al. (2014). Metformin and Male Reproduction: Effects on Sertoli Cell Metabolism. Br. J. Pharmacol. 171 (4), 1033–1042. 10.1111/bph.12522 - DOI - PMC - PubMed
    1. Chen Q., Vazquez E. J., Moghaddas S., Hoppel C. L., Lesnefsky E. J. (2003). Production of Reactive Oxygen Species by Mitochondria. J. Biol. Chem. 278 (38), 36027–36031. 10.1074/jbc.M304854200 - DOI - PubMed
    1. Chiarelli R., Martino C., Agnello M., Bosco L., Roccheri M. C. (2016). Autophagy as a Defense Strategy Against Stress: Focus on Paracentrotus lividus Sea Urchin Embryos Exposed to Cadmium. Cell Stress and Chaperones 21 (1), 19–27. 10.1007/s12192-015-0639-3 - DOI - PMC - PubMed
    1. Crisóstomo L., Alves M. G., Gorga A., Sousa M., Riera M. F., Galardo M. N., et al. (2018). Molecular Mechanisms and Signaling Pathways Involved in the Nutritional Support of Spermatogenesis by Sertoli Cells. Methods Mol. Biol. 1748, 129–155. 10.1007/978-1-4939-7698-0_11 - DOI - PubMed

LinkOut - more resources