Objective To explore the association of testosterone (T), testosterone receptor (AR), and heat shock protein 90 (HSP90) levels in the peripheral blood with those in the testicular tissue of azoospermia patients. Methods A total of 45 azoospermia patients were selected in this study. Their blood levels of luteotropic hormone (LH), follicle-stimulating hormone (FSH), T, AR, and HSP90 were measured. The testicular tissues were biopsied to observe the presence of sperm under microscope. Then the biopsied tissues were ground, centrifuged to obtain the supernatant, which was then subjected to detection of T, AR, and HSP90 levels. According to the biopsied results and FSH value, the patients were divided into three groups: group A (had sperm, FSH<8 IU/L), B (no sperm, FSH<8 IU/L), and C (no sperm, FSH>8 IU/L). The blood and testicular levels of T, AR, and HSP90 were compared between the three groups, and the relationship between FSH, LH, T, AR, and HSP90 levels was analyzed. Results There were significant differences between the three groups regarding the levels of AR (F=5.478,P=0.026) and HSP90 (F=5.972,P=0.019) in the blood samples, and levels of T (F=6.757,P=0.006) and AR (F=4.988,P=0.047) in the testis tissue samples. We also found that FSH level was positively correlated with blood HSP90 level (r=0.729,P=0.001) and testicular T level (r=0.722,P=0.002); LH level was positively correlated with blood (r=0.629,P=0.012) level and testicular T levels (r=0.610,P=0.016); blood T level was negatively correlated with blood AR level (r=-0.665,P=0.005) while testicular T level was positively correlated with testicular AR level (r=0.544,P=0.029); and blood (r=0.559,P=0.042) and testicular (r=0.803,P=0.000) AR levels were positively correlated with their corresponding HSP90 levels. Conclusion Azoospermia patients with normal testicular volume have increased blood HSP90 and AR levels, and it has a feedback effect upon FSH and LH levels, with gradually increased testicular T and AR levels and unchanged HSP90 level.