Busulfan, an indispensable medicine in cancer treatment, can cause serious reproductive system damage to males as a side effect of its otherwise excellent therapeutic results. Its widespread use has also caused its accumulation in the environment and subsequent ecotoxicology effects. As a Chinese medicine, Wulingzhi (WLZ) has the effects of promoting blood circulation and improving female reproductive function. However, the potential effects of WLZ in male reproduction and in counteracting busulfan-induced testis damage, as well as its probable mechanisms, are still ambiguous. In this study, busulfan was introduced in a mouse model to evaluate its production of the testicular damage. The components of different WLZ extracts were compared using an untargeted metabolome to select extracts with greater efficacy, which were further confirmed in vivo. Here, we demonstrate abnormal spermatogenesis and low sperm quality in busulfan-injured testes. The WLZ extracts showed a strong potential to rehabilitate the male reproductive system; this effect was more prominent in room-temperature extracts. Additionally, both water and ethanol WLZ extracts at room temperature alleviated various busulfan-induced adverse effects. In particular, WLZ recovered spermatogenesis, re-activated arginine biosynthesis, and alleviated the increased oxidative stress and inflammation in the testis, ultimately reversing the busulfan-induced testicular injury. Collectively, these results suggest a promising approach to protecting the male reproductive system from busulfan-induced adverse side effects, as well as those of other similar anti-cancer drugs.

2-Bromopalmitate (2-BP) is a palmitoylation inhibitor that can prevent the binding of palmitic acid to proteins, thereby exhibiting significant effects in promoting inflammation and regulating the immune system. However, limited research has been conducted regarding the direct effects of 2-BP on the animal organism. Therefore, we probed mice injected with 2-BP for altered expression of inflammatory genes, with a focus on demonstrating changes in the intestinal flora as well as damage to the reproductive system. Our findings indicate that 2-BP can induce substantial inflammatory responses in visceral organs and cause testicular dysfunction. The key changes in the gut microbiota were characterized by an abundance of Firmicutes, Clostridiales, Rikenellaceae_RC9_gut_group, Desulfovibrio, Muribaculaceae, and Alistipes, and their metabolism has been intricately linked to visceral inflammation. Overall, the findings of this study provide a sound scientific basis for understanding the impact of high doses of 2-BP in mammals, while also offering crucial support for the development of preclinical models to suppress palmitoylation.

Wulingzhi (WLZ), a traditional Chinese medicine with diverse chemical constituents, has shown antioxidant and cytoprotective activities; however, its potential to protect spermatogonial stem cells against chemotherapy-induced oxidative injury remains unclear. In this study, a busulfan-induced oxidative injury model was established in porcine spermatogonial stem cells (PSSCs) to investigate the protective effects of WLZ and explore potential bioactive compounds and associated molecular mechanisms by integrating ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS)-based chemical profiling, network pharmacology, molecular docking, and in vitro experiments. Chemical profiling of WLZ by UHPLC–MS/MS, followed by integration with constituents retrieved from BATMAN-TCM and HERB, established an integrated library of 89 candidate compounds. Network pharmacology and molecular docking computationally prioritized ursolic acid (UA) as a candidate WLZ-associated constituent and AKT1 as a candidate regulatory target for subsequent experimental evaluation. Experimental evaluation showed that both WLZ and UA attenuated busulfan-induced oxidative injury, as evidenced by improved cell viability, reduced ROS accumulation, and restored antioxidant capacity. The protective effects of UA were associated with changes in AKT1 transcript expression and were attenuated by the AKT inhibitor Miransertib. In contrast, the PI3K–AKT pathway agonist Recilisib independently improved antioxidant-related parameters in busulfan-exposed PSSCs in the absence of UA. UA treatment was also accompanied by increased transcript levels of Nrf2 and the antioxidant genes SOD, CAT, and GPX1. These findings indicate that UA, prioritized from the integrated WLZ candidate compound library, independently attenuates busulfan-induced oxidative injury in PSSCs and suggest a possible involvement of AKT-related signaling in the observed antioxidant responses. This study provides preliminary in vitro evidence that WLZ and UA attenuate busulfan-induced oxidative injury in PSSCs and supports further investigation of their potential protective effects against chemotherapy-associated male reproductive toxicity.

RNA editing is increasingly recognized as a post-transcriptional modification that directly affects viral infection by regulating RNA stability and recoding proteins. the duck hepatitis A virus genotype 3 (DHAV-3) infection is seriously detrimental to the Asian duck industry. However, the landscape and roles of RNA editing in the susceptibility and resistance of Pekin ducks to DHAV-3 remain unclear. Here, we profiled dynamic RNA editing events in liver tissue and investigated their potential functions during DHAV-3 infection in Pekin ducks. We identified 11,067 informative RNA editing sites in liver tissue from DHAV-3-susceptible and -resistant ducklings at three time points during virus infection. Differential RNA editing sites (DRESs) between S and R ducks were dynamically changed during infection, which were enriched in genes associated with vesicle-mediated transport and immune-related pathways. Moreover, we predicted and experimentally verified that RNA editing events in 3′-UTR could result in loss or gain of miRNA–mRNA interactions, thereby changing the expression of target genes. We also found a few DRESs in coding sequences (CDSs) that altered the amino acid sequences of several proteins that were vital for viral infection. Taken together, these data suggest that dynamic RNA editing has significant potential to tune physiological processes in response to virus infection in Pekin ducks, thus contributing to host differential susceptibility to DHAV-3.