Qiangjing tablets ameliorate asthenozoospermia via mitochondrial ubiquitination and mitophagy mediated by LKB1/AMPK/ULK1 signaling

Abstract

Context

Therapeutic effects of Qiangjing tablets (QJT) on sperm vitality and asthenozoospermia (AZS) have been confirmed. However, the mechanism of action remains unclear.

Objective

This study investigates the effects of QJT on AZS and the underlying mechanism of action.

Materials and methods

Sixty Sprague–Dawley rats were randomly divided into six groups: Control, ORN (ornidazole; 200 mg/kg), ORN + QJT-low (0.17 g/mL), ORN + QJT-middle (0.33 g/mL), ORN + QJT-high (0.67 g/mL), and ORN + QJT + Radicicol (0.67 g/mL QJT and 20 mg/kg radicicol) groups. Pathological evaluation and analysis of mitophagy were conducted by H&E staining and transmission electron microscopy, respectively. Reactive oxygen species were detected by flow cytometry. Protein expression was determined by Western blotting.

Results

QJT significantly improved ORN-treated sperm motility and kinematic parameters, as well as the pathological symptoms of testicular and epididymal tissues. In particular, QJT mitigated impaired mitochondrial morphology, and increased the PHB, Beclin-1, LC3-II protein, and ROS levels (p < 0.05), and reduced the protein expression levels of LC3-I and p62 (p < 0.05). Mechanistically, QJT antagonized the downregulation of SCF and Parkin protein levels (p < 0.05). Furthermore, QJT significantly increased the protein expressions levels of LKB1, AMPKα, p-AMPKα, ULK1 and p-ULK1 (p < 0.05). The ameliorative effect of QJT on pathological manifestations, mitochondrial morphology, and the expressions of mitophagy and mitochondrial ubiquitination-related proteins was counteracted by radicicol.

Discussion and conclusions

QJT improved AZS via mitochondrial ubiquitination and mitophagy mediated by the LKB1/AMPK/ULK1 signaling pathway. Our study provides a theoretical basis for the treatment of AZS and male infertility.

Keywords:

• Mitophagy
• LKB1/AMPK/ULK1
• sperm motility
• mitochondria
• traditional Chinese medicine

Authors’ contributions

Approval of final manuscript: All authors. GL, JC, YX, YL, DC, PZ, ZM, DC, YY, and XH conceptualized the study, methodology, software, and data curation. GL and JC writing-original draft preparation, visualization, investigation, and supervision. GL and JC validation, writing-reviewing, and editing.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This research was funded by Science and Technology Development Fund of The Affiliated Hospital of Chengdu University of Traditional Chinese Medicine [18PY25]; "Xinglin Scholar" Scientific Research Promotion Plan of Chengdu University of Traditional Chinese Medicine [QNXZ2019013]; Special research project of Sichuan Administration of Traditional Chinese Medicine [2021MS474].