The global infertility rate has been increasing annually, and male infertility becoming severe. The proportion of idiopathic infertility is high, with most patients with ambiguous clinical symptoms. These symptoms manifested as declining semen quality, and the effects of autoimmune factors on this measure of fertility cannot be ignored. Aside from anti-sperm antibody for autoimmune infertility, detection indexes for male infertility are lacking. We successfully collected seminal fluid via electric stimulation from an experimental model of autoimmune orchitis in rats. We used the most advanced isobaric tags for relative and absolute quantitative proteomics analysis to analyze the seminal plasma of autoimmune infertility.