Arachidonic acid impairs the function of the blood-testis barrier via triggering mitochondrial complex-ROS-P38 MAPK axis in hyperthermal Sertoli cells

The death of Sertoli cells (SCs) under condition of heat stress (HS) affects spermatogenesis and is associated with impaired function of the blood-testis barrier (BTB). The fatty acid arachidonic acid (AA) is essential for the maintenance of cellular function. However, excessive release of AA during HS may adversely affect the repro-ductive function. The molecular mechanisms through which AA modulates the BTB in SCs are unclear. In this study, we found that 100 mu M AA damaged testicular morphology and accelerated SC apoptosis during HS, reducing the stability of tight junction proteins (TJPs), shown by measurement of the levels of Claudin 11, 5, Occludin, and trans-epithelial electrical resistance (TEER). It was also found that AA adversely affected TJPs by increasing the accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA), activating p38 mitogen-activated protein kinases (P38 MAPK) and reducing mitochondria DNA (mtDNA) and the expression of mitochondrial complexes I and III. In contrast, pretreatment with SB203508 (a P38 MAPK inhibitor), Rotenone (an inhibitor of complex I) and Antimycin A1 (an inhibitor of complex III) reversed TJPs degradation induced by AA. Interestingly, pretreatment of cells with 10 mu M Baicalein, a 12/15 lipoxygenase (12/15-LOX)-dependent inhibitor of AA production, protected against AA-induced TJPs degradation, restored mitochondrial function, and reduced apoptosis. These results suggested an intriguing link between the induction of TJPs degradation induced by AA overload and mitochondrial antioxidant function during HS, which was found to be regulated by the mitochondrial complex-ROS-P38 MAPK axis.

Automated summary

The death of Sertoli cells (SCs) under heat stress (HS) affects spermatogenesis and the blood-testis barrier (BTB). Excessive release of arachidonic acid (AA) during HS damages testicular morphology and accelerates SC apoptosis, reducing the stability of tight junction proteins (TJPs). AA adversely affects TJPs through the accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA), activation of p38 mitogen-activated protein kinases (P38 MAPK), and reduction of mitochondrial function. However, pretreatment with inhibitors reversed the degradation of TJPs and protected against AA-induced damage, suggesting the involvement of the mitochondrial complex-ROS-P38 MAPK axis.