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.
Highlights
Sertoli cells (SCs), which are sustentacular cells in the mammalian testicular seminiferous tubule, play critical roles in the maintenance and regulation of spermatogenesis and provide a favorable microenvironment for sperm development (Griswold, 1998)
Our results suggested that BPFL has adverse effects in porcine SCs, leading to oxidative stress, mitochondriaER dysfunction, DNA damage and apoptosis, and that Chlorogenic acid (CA) supplementation has potential protective effects against the impairments induced by BPFL exposure
As increased oxidative stress was observed in BPFL-treated SCs, we argued that BPFL may induce an autophagic mode of cell death, but no significant increase in light chain 3 (LC3) expression was observed in the 50 μM-treated cells compared to the control group, indicating that the effect of BPFL on autophagy of SCs was weak
Summary
Sertoli cells (SCs), which are sustentacular cells in the mammalian testicular seminiferous tubule, play critical roles in the maintenance and regulation of spermatogenesis and provide a favorable microenvironment for sperm development (Griswold, 1998). Studies in different models of SC damage, such as male Fischer rats and mice, suggest that spermatogenesis is vulnerable to disruption when SCs are damaged and that targeting key SC functions can lead to rapid and massive germ cell death (Richburg and Boekelheide, 1996; Hasegawa et al, 1997). SCs cultured in vitro are often used as a cell model for studying the testicular toxicity of drugs and compounds (Alves et al, 2014; Zhao et al, 2021)
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