The present study was aimed to discuss the role and clinical significance of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the deterioration of chronic hepatitis B patients superinfected with acute hepatitis E. Serum TNF-α and IL-6 in chronic hepatitis B patients superinfected with acute hepatitis E (n=30), chronic hepatitis B patients (n=20), acute hepatitis E patients(n=20), and healthy volunteers (n=20) were detected by enzyme-linked immunosorbent assay (ELISA). Main biochemical quantitative analyses of serum levels of alanine aminotransferase (ALT), aspartamine transferase (AST) and total bilirubin (TBIL), cholesterol ester (CHE), albumin (ALB),and prothrombin activity (PTA) were detected by automatic biochemical analyzer. Compared to that of chronic hepatitis B patients, acute hepatitis E patients, and healthy volunteers, the serum levels of TNF-α and IL-6 in chronic hepatitis B patients superinfected with acute hepatitis E were significantly increased (P <0.01). Compared to that of chronic hepatitis B patients and acute hepatitis E patients, the serum levels of ALT, AST and TBIL in chronic hepatitis B patients superinfected with acute hepatitis E were significantly increased (P <0.05), the serum levels of CHE, ALB and PTA reduced obviously (P <0.05). The serum levels of TNF-α and IL-6 in chronic hepatitis B patients superinfected with acute hepatitis E showed a significant positive correlation with ALT and TBIL and a significant negative correlation with PTA (P <0.05). TNF-α and IL-6 participate in the process of immune injury in chronic hepatitis B patients superinfected with acute hepatitis E. Detection of the serum levels of TNF-α and IL-6 has practical clinical value in determining the degree of liver injury and prognosis of chronic hepatitis B patients superinfected with acute hepatitis E.