Additional file 1 of The gut metabolite 3-hydroxyphenylacetic acid rejuvenates spermatogenic dysfunction in aged mice through GPX4-mediated ferroptosis

Additional file 1: Figure S1. The sperm motility parameters of all FMT mice. Figure S2. Sperm quality and spermatogenesis of young and old donor mice. Figure S3. Different microbiota between young and old mice. Figure S4. Characterization of metabolites of cecum feces in young and old donor mice. Figure S5. Spearman correlation analyses between gut microbiota and differentiated gut metabolites. Figure S6. KEGG pathway analysis of the significantly regulated metabolites between groups in the testis tissues of the y FMT o and o FMT o groups. Figure S7. Sperm motility parameters of 3-HPAA-treated-old mice. Figure S8. Ferroptosis-related protein expression of o FMT y and y FMT o mice. Figure S9. Expression and knockdown efficiency of GPX4 siRNA. Table S1. PCR primer sequences. Table S2. GPX4 siRNA nucleotide sequences

Additional file 2 of The gut metabolite 3-hydroxyphenylacetic acid rejuvenates spermatogenic dysfunction in aged mice through GPX4-mediated ferroptosis

Additional file 2: Data S1. Non-targeted metabolome in the microbiome, plasma, and testis samples of donor or FMT mice

Additional file 4 of The gut metabolite 3-hydroxyphenylacetic acid rejuvenates spermatogenic dysfunction in aged mice through GPX4-mediated ferroptosis

Additional file 4: Data S3. RNA sequencing of the testis tissues of the 3-HPAA and vehicle mice

Additional file 3 of The gut metabolite 3-hydroxyphenylacetic acid rejuvenates spermatogenic dysfunction in aged mice through GPX4-mediated ferroptosis

Additional file 3: Data S2. Targeted metabolome in the plasma and testis of the 3-HPAA and vehicle mice

Hepatitis E virus infects human testicular tissue and Sertoli cells

Globally, hepatitis E virus (HEV) infections are prevalent. The finding of high viral loads and persistent viral shedding in ejaculate suggests that HEV replicates within the human male genital tract, but its target organ is unknown and appropriate models are lacking. We aimed to determine the HEV tropism in the human testis and its potential influence on male reproductive health. We conducted an ex vivo culture of human testis explants and in vitro culture of primary human Sertoli cells. Clinically derived HEV genotype 1 (HEV1) and HEV3 virions, as well as rat-derived HEV-C1, were used for inoculation. Transcriptomic analysis was performed on testis tissues collected from tacrolimus-treated rabbits with chronic HEV3 infection. Our findings reveal that HEV3, but not HEV1 or HEV-C1, can replicate in human testis explants and primary human Sertoli cells. Tacrolimus treatment significantly enhanced the replication efficiency of HEV3 in testis explants and enabled successful HEV1 infection in Sertoli cells. HEV3 infection disrupted the secretion of several soluble factors and altered the cytokine microenvironment within primary human Sertoli cells. Finally, intratesticular transcriptomic analysis of immunocompromised rabbits with chronic HEV infection indicated downregulation of genes associated with spermatogenesis. HEV can infect the human testicular tissues and Sertoli cells, with increased replication efficiency when exposed to tacrolimus treatment. These findings shed light on how HEV may persist in the ejaculate of patients with chronic hepatitis E and provide valuable ex vivo tools for studying countermeasures.</p