The aim of the study is to investigate the effects of sub-chronic poisoning with arsenic on the testes of chickens treated. Seventy-two 1-day-old chickens were randomly divided into 4 groups and provided food with different doses arsenic. The histological changes were examined. The mRNA levels of inflammatory factors, including transcription factor nuclear factor-κB (NF-κB), tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), prostaglandin E synthase (PTGEs), and heat shock proteins (Hsps), including Hsp70, Hsp90, Hsp60, Hsp40, and Hsp27 were assessed by quantitative real-time PCR in the testes of chickens. The protein expressions of iNOS, Hsp60, and Hsp70 were detected by western blot. Increased mRNA and protein levels of inflammatory factors and Hsps with testicular damage showed that arsenic-induced testicular toxicity includes inflammatory and heat shock response in chickens, and that increased Hsps levels may play a protective role in inflammation damage induced by arsenic on the testes of chickens.