Objective: To analyze the expression levels of multiple serum biochemical and immune indicators in patients with chronic heart failure, explore the correlation between each indicator and the severity of the patient’s condition, and screen objective serological reference indicators that can be used to evaluate the grade and prognostic risk of heart failure. Methods: The research investigation period was from October 2024 to February 2026. The selected research subjects were 126 patients with chronic heart failure who were treated in our hospital. According to the New York Heart Association (NYHA) cardiac function classification standards, the patients were divided into the grade II group with 42 cases, the grade III group with 45 cases, and the grade IV group with 39 cases. In addition, 60 healthy people who underwent physical examination in our hospital during the same period were selected as the control group. Venous blood was collected from all study subjects on an empty stomach to detect serum uric acid (UA), creatinine (Scr), urea nitrogen (BUN), high-sensitivity C-reactive protein (hs-CRP), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) levels. The differences in indicators between heart failure patients with different cardiac function grades and healthy people were compared. Pearson correlation was used to analyze the relationship between each serum indicator and the severity of heart failure. Results: The serum UA, Scr, BUN, hs-CRP, TNF-α, and IL-6 levels of the patients in the observation group were all higher than those of the control group (P < 0.05); as the NYHA cardiac function class increased, the levels of various serum biochemical and immune-inflammatory indicators in the patients increased in a stepwise manner. The comparison difference value between grade II, grade III, and grade IV groups was P<0.05; Pearson correlation analysis results showed that serum UA, Scr, BUN, hs-CRP, TNF-α, and IL-6 levels were positively correlated with the severity of chronic heart failure. Conclusion: A number of serum biochemical and immune-inflammatory indicators are abnormally highly expressed in patients with chronic heart failure, and the expression levels of the indicators increase as the patient's cardiac function injury worsens, and are closely related to the severity of the disease. Clinically, the above serum indicators can be jointly detected to objectively evaluate the progression of heart failure patients, providing a reliable laboratory basis for clinical hierarchical diagnosis and treatment, intervention plan adjustment, and prognosis assessment.