By predicting and elucidating the multi-component, multi-target, and multi-pathway therapeutic effects of MLST on varicocele-associated male infertility through network analysis, it was experimentally verified that MLST can inhibit the activation of the PI3K/Akt/mTOR signaling pathway, reduce the expression of HIF1α, and further attenuate VC-induced oxidative stress and apoptosis in the testis. This evidence concerns the gene PIK3CA and varicocele.