Plant-derived exosome-like nanovesicles improve testicular injury by alleviating cell cycle arrest in Sertoli cells

The testis plays a vital role in male reproduction, and its function directly impacts spermatogenesis and male fertility. Cyclophosphamide, a commonly used chemotherapeutic agent, induces severe testicular toxicity; however, effective strategies for prevention or treatment are currently lacking. In this study, we isolated and identified exosome-like nanovesicles derived from Cistanche deserticola (CDELNs), characterized their composition, and investigated their therapeutic effects and molecular mechanisms on cyclophosphamide-induced testicular injury. Specifically, we found that CDELNs are preferentially taken up by testicular Sertoli cells, and this uptake process is mediated by heparan sulfate proteoglycans (HSPG). Mechanistically, miR159b-3p derived from CDELNs alleviates cell cycle arrest and restores testicular function by inhibiting the expression of the cell cycle inhibitor P21, thereby promoting the phosphorylation-dependent activation of cyclin-dependent kinase 1 (CDK1). Furthermore, single-cell transcriptomic analysis of testicular tissues from patients with non-obstructive azoospermia (NOA) in published database revealed that Sertoli cells and P21 are critically involved in male reproductive disorders, suggesting that CDELNs-based intervention strategy targeting P21 holds broad prospects for clinical translation. Collectively, our study reveals firstly that CDELNs, a novel bioactive substrate of Cistanche deserticola, exert therapeutic effects on male testicular injury by regulating the cell cycle pathway through their miRNA.