Yang Fu,1,2 Peipei Yuan,1,2 Manyv Wang,1 Yajuan Zheng,1 Yan Zhang,1 Lirui Zhao,1 Qingyun Ma,1 Pengsheng Wang,1 Xiaotian Sun,1 Xiaoke Zheng,1,2 Weisheng Feng1,2 

1Department of Pharmacy, Henan University of Chinese Medicine, Zhengzhou 450046, China, Zhengzhou, 450046, People’s Republic of China; 2The Engineering and Technology Center for Chinese Medicine Development of Henan Province, Zhengzhou, 450046, People’s Republic of China

Correspondence: Xiaoke Zheng; Weisheng Feng, Department of Pharmacy, Henan University of Chinese Medicine, The Engineering and Technology Center for Chinese Medicine Development of Henan Province, 156 Jinshui East Road, Zhengzhou, 450046, People’s Republic of China, Email zhengxk.2006@163.com; fwsh@hactcm.edu.cn

Purpose: Doxorubicin (DOX) precipitates cell apoptosis in testicular tissues, and it is imperative to develop drugs to alleviate the spermatogenic disorders it causes. Eugenia caryophyllata Thunb is often used to treat male sexual disorders. Eugenol, a major component of Eugenia caryophyllata Thunb. has inadequate stability and low solubility, which limits its pharmacological effects. Eugenol nanoparticles (NPs) (ENPs) are expected to overcome these limitations. The protective effects of ENPs against DOX-induced reproductive toxicity were studied in mice.
Methods: Eugenol was encapsulated in Methoxy-Poly(ethylene glycol)-Poly(lactide-co-glycolide) nanoparticles (mPEG-PLGA-NPs), and their role in ameliorating spermatogenic dysfunction was verified in vivo and in vitro.
Results: We present a promising delivery system that encapsulates eugenol into mPEG-PLGA-NPs and forms them into nanocomposites. In vitro, ENPs significantly reduced doxorubicin-induced ROS and inflammatory factors in GC-1 cells and regulated the expression of the mitochondrial autophagy protein PINK1 and meiosis-related protein SCP3. In vivo, ENPs significantly increased sperm motility in mice, reduced apoptosis and oxidative stress in the testes, inhibited the testicular PINK1/Parkin and BNIP3/NIX signaling pathways, and enhanced the expression of factors associated with meiosis.
Conclusion: Given their safety and efficacy, these ENPs have potential application prospects in mitigating doxorubicin-induced spermatogenic dysfunction.

Keywords: Eugenol nanoparticles, Doxorubicin, spermatogenic dysfunction, PINK1/Parkin and BNIP3/NIX signaling pathways, Mitochondrial autophagy