Implication of Polyhistidine, a Novel Apoptosis Inhibitor, in Inhibiting Lipopolysaccharide-Induced Apoptosis in Boar Sperm

SIMPLE SUMMARY: Gram-negative bacteria are the main pathogenic microorganisms found in human and animal semen. Lipopolysaccharide (LPS), a component of the cell wall of Gram-negative bacteria, has been linked to inducing apoptosis in human and rat sperm; however, little is known regarding LPS-induced apoptosis in boar sperm. This detrimental effect of LPS is potentially mediated via competitive bidding with toll-like receptor (TLR) 4 on the cytoplasmic membrane. Therefore, it is reasonable to elucidate the potential mechanisms by which the binding of LPS and TLR4 could be prevented. Polyhistidine is widely used for the delivery of nucleic acids and antibodies into the cell cytoplasm, and it is a novel TLR4 agonist. In the current study, we envisaged that pHis might also serve as an effective tool for inhibiting LPS-induced apoptosis in boar sperm. The new finding of our present study is that pHis could inhibit, to some extent, LPS-induced boar sperm apoptosis, and it could ameliorate the overall sperm quality parameters under liquid storage or at 37 °C incubation conditions. However, further investigation should be continued to fully elucidate the mechanistic basis of these ameliorative effects of pHis. ABSTRACT: Lipopolysaccharide (LPS) released from Gram-negative bacteria binds to toll-like receptor 4 (TLR4) and induces boar sperm apoptosis. Similarly, polyhistidine (pHis), a TLR4 agonist, can also bind to TLR4. We hypothesized that pHis could inhibit LPS-induced sperm apoptosis by competitively binding to TLR4 to then improve sperm quality. Therefore, the objective of this study was to examine whether pHis can inhibit LPS-induced sperm apoptosis and affect sperm quality. The results showed that the concentrations of bacterial colonies were significantly increased from 36 to 120 h under liquid storage conditions (p < 0.05); however, concentrations of LPS in boar semen showed a relatively constant trend (4.98 ± 1.55 EU/mL) following 120 h storage. The addition of 100 μg/mL pHis in the BTS extender significantly improved boar sperm motility and viability at 37 °C, and it significantly (p < 0.05) inhibited boar sperm apoptosis under liquid storage (17 °C) and at 37 °C incubation conditions. The co-treatment of LPS and pHis further confirmed that pHis played its role in inhibiting LPS-induced sperm apoptosis. In conclusion, our preliminary findings provide reasonable evidence that pHis could act as an inhibitor of LPS-induced apoptosis in boar sperm stored for longer periods of time. pHis might inhibit LPS-induced sperm apoptosis by competitively binding to TLR4. Nevertheless, further mechanistic studies are awaited to fully elucidate its potential implication in inhibiting LSP-induced apoptosis.