Male fertility has been shown to be dependent on cholesterol homeostasis. This lipid is essential for testosterone synthesis and spermatogenesis, but its levels must be maintained in an optimal range for proper testicular function. In particular, sperm cells development is very sensitive to high cholesterol levels, noticeably during acrosomal formation. The aim of this work was to study whether the molecular pathway that regulates intracellular cholesterol, the Sterol Regulatory Element-Binding Protein (SREBP) pathway, is affected in the testicles of animals under a fat diet. To investigate this, we took advantage of the non-obese hypercholesterolemia (HC) model in New Zealand rabbits that displays poor sperm and seminal quality. The testicular expression of SREBP isoform 2 (SREBP2) and its target molecules 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGCR) and low-density lipoprotein receptor (LDLR) were studied under acute (6 months) and chronic (more than 12 months) fat intake by RT-PCR, western blot and immunofluorescence. Our findings showed that fat consumption promoted down regulation of the SREBP2 pathway in the testicle at 6 months, but upregulation after a chronic period. This was consistent with load of testicular cholesterol, assessed by filipin staining. In conclusion, the intracellular pathway that regulates cholesterol levels in the testicle is sensitive to dietary fats, and behaves differently depending on the duration of consumption: it has a short-term protective effect, but became deregulated in the long term, ultimately leading to a detrimental situation. These results will contribute to the understanding of the basic mechanisms of the effect of fat consumption in humans with idiopathic infertility.

Background Rabbits are sensitive to dietary cholesterol and rapidly develop hypercholesterolemia, leading to prominent subfertility. Sterol regulatory element‐binding protein isoform 2 drives the intracellular cholesterol pathway in many tissues, including the testicles. Its abnormal regulation could be the mainly responsible for the failure of suppressing cholesterol synthesis in a cholesterol‐enriched environment, ultimately leading to testicular and seminal alterations. However, extra‐virgin olive oil consumption has beneficial properties that promote lowering of cholesterol levels, including the recovery of seminal parameters altered under a high‐fat diet. Objectives Our goal was to investigate the effects of high‐fat diet supplementation with extra‐virgin olive oil at the molecular level on rabbit testes, by analyzing sterol regulatory element‐binding protein isoform 2 protein and its corresponding downstream effectors. Materials and methods During 12 months, male rabbits were fed a control diet, high‐fat diet, or 6‐month high‐fat diet followed by 6‐month high‐fat diet plus extra‐virgin olive oil. Serum lipids, testosterone levels, bodyweight, and seminal parameters were tested. The mRNA and protein levels of sterol regulatory element‐binding protein isoform 2, 3‐hydroxy‐3‐methyl‐glutaryl‐coenzyme A reductase, and low‐density lipoprotein receptor were determined by semi‐quantitative polymerase chain reaction and Western blotting techniques. The expression pattern of sterol regulatory element‐binding protein isoform 2 protein in the rabbit testicles was studied by indirect immunofluorescence. In addition, testicular cholesterol was detected and quantified by filipin staining and gas chromatography. Results The data showed that the addition of extra‐virgin olive oil to high‐fat diet reduced testicular cholesterol levels and recovered the expression of sterol regulatory element‐binding protein isoform 2, 3‐hydroxy‐3‐methyl‐glutaryl‐coenzyme A reductase, and low‐density lipoprotein receptor initially altered by the high‐fat diet. Discussion and conclusions The combination of high‐fat diet with extra‐virgin olive oil encourages testicular recovery by modifying the expression of the enzymes related to intracellular cholesterol management.