Exploring Pathogenesis and Biomarkers Through Establishment of a Rat Model of Male Infertility with Liver Depression and Kidney Deficiency

Abstract

Abstract Objectives To establish a rat model that accurately replicates the clinical characteristics of Liver Depression and Kidney Deficiency (LD & KD) in male infertility (MI) and investigate the pathogenesis. Methods After subjecting the rats to chronic restraint stress (CRS) and adenine treatment, a series of tests were conducted, including ethological assessments, evaluations of reproductive characteristics, measurements of biochemical parameters, histopathological examinations, and analyses of urinary metabolites. Results Compared to the control, the model exhibited significant manifestations of LD & KD in MI, including reduced responsiveness, diminished frequency of capturing estrous female rats, and absence of mounting behavior. Additionally, the kidney coefficient increased markedly, while the coefficients of the testis and epididymis decreased significantly. Sperm counts and viabilities decreased notably, accompanied by an increase in sperm abnormalities. Dysregulation of reproductive hormone levels in the serum was observed, accompanied by an upregulation of proinflammatory cytokines expressions in the liver and kidney, as well as exacerbated oxidative stress in the penile corpus cavernosum and testis. The seminiferous tubules in the testis exhibited a loose arrangement, loss of germ cells, and infiltration of inflammatory cells. Furthermore, utilizing urinary metabolomics and bioinformatics analysis, 5 key biomarkers and 2 crucial targets most closely linked to MI were revealed. Conclusion The study successfully established a clinically relevant animal model of LD & KD in MI. It elucidates the pathogenesis of the condition, identifies key biomarkers and targets, and provides a robust scientific foundation for the prediction, diagnosis, and treatment of LD & KD in MI.