Purpose: To compare the microenvironment change of three type thermal ablation: transperineal laser ablation (TPLA), radiofrequency ablation (RFA) and microwave ablation (MWA) application canine prostate and their clinical significances. Methods: Nine beagles were randomly assigned to TPLA, RFA and MWA group. The ablation was operated in prostate. The level of cytokine (TNF-α, IL-12, TGF-β1 and IL-10) in serum was detected at four time point (before ablation, immediately, one week and one month after ablation). The positive cells of macrophages in the border of ablation lesions were detected. Results: One week after ablation, the levels of pro-inflammatory cytokine (TNF-α) in the TPLA group was the highest, followed by the RFA group, and the MWA group was the lowest (159.11 ± 53.26 pg/ml, 129.57 ± 23.42 pg/ml, 53.65 ± 21.34 pg/ml, P<0.03). However, one month later, the level of TNF-α had a continuous increase in TPLA group while which showed a significant decrease trend in other groups. The levels of anti-inflammatory cytokine (TGF-β1) in the TPLA group was significantly lower than other two groups (48.92 ± 48.67 pg/ml, 218.85 ± 27.56 pg/ml, 167.67 ± 45.67 pg/ml, P<0.05). M1-like macrophages were dominant in TPLA group, and M2-like macrophages were dominant in RFA and MWA groups. Conclusion: Microenvironment changes in the TPLA group were mainly pro-inflammatory, while anti-inflammatory responses were observed in the RFA and MWA groups.