Background: Mangiferin (MA), a bioactive C-glucosyl xanthone with a wide range of interesting therapeutic properties, has recently attracted considerable attention. However, its application in biomedicine is limited by poor solubility and bioavailability. Carbon dots (CDs), novel nanomaterials, have immense promise as carriers for improving the biopharmaceutical properties of active components because of their outstanding characteristics. Methods: In this study, a novel water-soluble carbon dot (MC-CDs) was prepared for the first time from an aqueous extract of Moutan Cortex Carbonisata, and characterized by various spectroscopies, zeta potential and high-resolution transmission electron microscopy (HRTEM). The toxicity effect was investigated using the CCK-8 assay in vitro. In addition, the potential of MC-CDs as carriers for improving the pharmacokinetic parameters was evaluated in vivo. Results: The results indicated that MC-CDs with a uniform spherical particle size of 1– 5 nm were successfully prepared, which significantly increased the solubility of MA in water. The MC-CDs exhibited low toxicity in HT-22 cells. Most importantly, the MC-CDs effectively affected the pharmacokinetic parameters of MA in normal rats. UPLC-MS analysis indicated that the area under the maximum blood concentration of MA from mangiferin-MC-CDs (MA-MC-CDs) was 1.6-fold higher than that from the MA suspension liquid (MA control) after oral administration at a dose of 20 mg/kg. Conclusion: Moutan Cortex-derived novel CDs exhibited superior performance in improving the solubility and bioavailability of MA. This study not only opens new possibilities for the future clinical application of MA but also provides evidence for the development of green biological carbon dots as a drug delivery system to improve the biopharmaceutical properties of insoluble drugs.

Background: Obesity-induced oligoasthenospermia is associated with testicular lipid metabolism. However, the mechanisms underlying the lipid homeostasis imbalance of obesity-induced oligoasthenospermia are unclear. Methods: Male C57BL/6 mice fed a high-fat diet were established for the in vivo model. TM3/TM4 cells were treated with palmitic acid (PA) in vitro. Proteomics analyzed differential proteins in the testis. The Oil red O and Nile red were used to observe lipid droplets (LDs). Filipin staining was used to observe free cholesterol (FC). Hormone-sensitive lipase (HSL), Low-density lipoprotein receptor (LDLr), Scavenger receptor class B type I (SR-BI), and ATP-binding cassette transporter A1 (ABCA1) expressions were analyzed using qRT-PCR, WB, Immunofluorescence, and immunohistochemistry. Testosterone synthesis and Blood-testis barrier (BTB) integrity were evaluated by ELISA, Transmission electron microscope, and WB. Results: HFD mice exhibited elevated blood lipid, reduced sperm quality, and hormonal imbalances. Meanwhile, testosterone synthesis was impaired, and BTB was damaged in HFD mice. In vivo and in vitro models, LDs deposition was observed, and HSL expression was down-regulated in Leydig and Sertoli cells. However, the expressions of cholesterol uptake receptors and efflux transporters, as well as the levels of FC, in the two cells were inconsistent. In Leydig cells, the expression of cholesterol uptake receptors (LDLr and SR-BI) was upregulated, resulting in increased FC levels. In Sertoli cells, cholesterol efflux transporter (ABCA1) expression was upregulated, and FC levels decreased. Overexpression of HSL ameliorated LDs accumulation and increased testosterone levels. Conclusion: The down-regulation of HSL and dysregulation of cholesterol receptors/transporter may affect lipid homeostasis, thereby damaging testicular function.

Background: Oligoasthenospermia is one of the main causes of male infertility. Researchers usually use chemical drugs to directly damage germ cells to prepare oligoasthenospermia models, which disregards the adhesion and migration between spermatogenic cells and Sertoli cells. TAp73 is a critical regulator of the adhesin of germ cell; thus, we sought to explore a novel oligoasthenospermia model based on TAp73 gene suppression.Methods: Mice in the Pifithrin‐α group were injected intraperitoneally with 2.5 mg/kg Pifithrin‐α (TAp73 inhibitor) daily for 30 consecutive days. Reproductive hormone levels and epididymal sperm quality, as well as the network morphology of Sertoli cells were tested.Results: Sperm density, motility, and the relative protein and mRNA expression of TAp73 and Nectin 2 were obviously decreased in the Pifithrin‐α group compared with the normal control group. No significant distinction was observed in the relative mRNA and protein expression of ZO‐1. Furthermore, the tight junctions (TJs) and apical ectoplasmic specialization (ES) were destroyed in the Pifithrin‐α group.Conclusion: The above results indicate that we successfully established a new oligoasthenospermia mouse model. This study provides a foundation for further exploration of the roles of TAp73 genes during spermatogenesis and provides new research objects for further oligospermia research and future drug discovery.

Objective: This study investigates the relationship between TAp73 protein overexpression and sperm quality, and evaluates the prophylactic and therapeutic effects of Wuzi-Yanzong-Wan (WZYZW) on oligoasthenospermia (OA) induced by tripterygium glycosides (TGs) and etoposide in rats. Furthermore, it elucidates the underlying mechanism by analyzing the intervention’s role in downregulating TAp73 protein expression and modulating the P38 MAPK-ADAM17 signaling pathway, ultimately providing empirical evidence to support its clinical application.Methods: Sprague-Dawley (SD) rats received TGs (40 mg/kg/d for 8 weeks) to induce OA. Intervention groups were treated with either WZYZW (1 or 2 g/kg/d) or Shengjing Capsule (SJJN). Meanwhile, a TAp73-overexpressing Sertoli-germ cell co-culture system was established using etoposide (200 μM for 48 h) and treated with WZYZW-containing serum (2.5%–10%), to explore the mechanisms associated with TAp73.Results: The in vivo experimental results demonstrated that WZYZW treatment significantly ameliorated testicular atrophy in model rats, evidenced by increased testicular volume and elevated testicular/epididymal indices. Simultaneously, WZYZW effectively reversed sperm quality impairment in the model group, manifesting as significantly increased sperm density, enhanced sperm motility, and reduced sperm abnormality rate. Furthermore, WZYZW treatment significantly upregulated serum levels of key sex hormones (e.g., testosterone, follicle-stimulating hormone, luteinizing hormone). Mechanistic investigations revealed that WZYZW markedly suppressed germ cell apoptosis (flow cytometry confirmed a significant decrease in apoptotic cell proportion) and attenuated DNA damage (indicated by significantly reduced γ-H2AX (a DNA damage marker) fluorescence intensity). WZYZW effectively restored impaired mitochondrial function and suppressed the expression of key proteins in the mitochondrial apoptotic pathway (e.g., decreased Bax/Bcl-2 ratio, reduced Cyt-c release, inhibited activation of Caspase-9 and Caspase-3). In vivo experimental results showed that WZYZW significantly inhibited cell apoptosis, effectively suppressed membrane potential depolarization in TAp73-overexpressing model cells, inhibited the opening of the mitochondrial permeability transition pore, and concurrently suppressed the expression of proteins associated with the mitochondrial apoptotic pathway. Collectively, these changes blocked the TAp73-p38 MAPK/ADAM17 axis-activated mitochondrial apoptotic cascade, thereby exerting its testicular protective effects.Conclusion: This study highlights that WZYZW effectively alleviates testicular DNA damage and mitochondrial apoptosis, suggesting that its mechanism may be associated with reduced TAp73 expression. These experimental findings provide a novel therapeutic target for the clinical application of WZYZW in OA treatment.