To detect the serum silent mating type information regulation 2 homolog 1 (SIRT1) and high mobility group box 1 (HMGB1) levels of the patients with ovarian cancer, and to analyze relationship between the parameters of contrast-enhanced ultrasound (CEUS), and the serum SIRT1 and HMGB1 levels of patients and their lymph node metastasis. Methods: The clinical data of a total of 96 patients with ovarian cancer from June 2016 to December 2019 were collected in study group, which included 42 patients with lymph node metastasis in group A and 54 patients without lymph node metastasis in group B. Another 96 women without ovarian cancer were included in control group. The quantitative analysis of contrastenhanced color ultrasound was performed on all of these patients. The levels of serum SIRT1 and HMGB1 were detected by enzyme linked immunosorbent assay (ELISA). The parameters of contrast-enhanced ultrasound of the patients were compared among these groups. The values of the parameters of quantitative analysis by contrast-enhanced ultrasound and the serum SIRT1 and HMGB1 levels for diagnosing the lymph node metastasis of ovarian cancer were analyzed. Results: There were no significant differences in the arrival time and perfusion time of contrast agent of the patients among the control group, group A, and group B (P＞0.05), but the peak time and basic intensity of contrast agent of the patients in group A and group B were significantly less than those of the patients in the control group. The enhanced intensity of contrast agent of the patients in group A significantly higher than that of the patients in group B, but the peak time and basic intensity of contrast agent of the patients in group A were significantly lower (all P＜0.05). The serum SIRT1 level of the patients in the control group, group B, and group A had decreased gradually, but the serum HMGB1 level of the patients in the control group, group B, and group A had increased gradually (P＜0.05). The analysis of receiver operator characteristic curve showed that the peak time and the basal intensity of the parameters by contrast-enhanced ultrasound, and the serum SIRT1 and HMGB1 levels had certain values for diagnosing the lymph node metastasis of ovarian cancer. The combination of the parameters by contrast-enhanced ultrasound and the serum SIRT1 and HMGB1 levels had the highest diagnosis value, which AUC, sensitivity, and specificity were 0.952, 92.6%, and 92.9%, respectively. Logistic regression analysis showed that the peak time and the basal intensity of the parameters by contrast-enhanced ultrasound, and the serum HMGB1 level were the risk factors of the lymph node metastasis of ovarian cancer, and the serum SIRT1 level was a protective factor of the lymph node metastasis of ovarian cancer. Conclusion: Quantitative analysis of contrast-enhanced ultrasound combined with the levels of serum SIRT1 and HMGB1 can improve the diagnostic efficiency of the lymph node metastasis of ovarian cancer, which can provide the evidences for evaluating the prognosis after clinical diagnosis and treatment.