ABSTRACT Purpose The present study aimed to determine the serum level of asprosin, a novel glucogenic and orexigenic hormone, in NAFLD and hypothyroidism patients. Methods This is a case–control study that included three groups of age and sex-matched subjects: hypothyroid patients with NAFLD (n = 26), hypothyroid patients without NAFLD (n = 25) and normal control subjects (n = 27). A detailed clinical history was obtained, and an examination was performed. Laboratory tests included fasting plasma glucose and insulin, lipid profile, TSH, free T4, AST, ALT, albumin, and asprosin. The homeostatic model assessment of insulin resistance (HOMA-IR) score was calculated. Abdominal ultrasonography was performed to calculate the hepatorenal index and define participants with NAFLD. The data were statistically analyzed. Results There was a statistically significant difference between the three groups regarding serum asprosin concentration (p < 0.01). Serum asprosin levels were significantly higher in the patients with hypothyroidism and NAFLD compared to both other groups [2.70 ng/mL (2.17–4.10), 1.26 ng/mL (0.96–2.50), and 1.72 ng/mL (1.41–2.12), respectively]. There was no significant difference between the other two groups. There was a significant positive correlation of moderate strength between asprosin and HOMA-IR, BMI, and waist circumference. Asprosin and HOMA-IR were significantly higher in patients with NAFLD compared to subjects with normal liver (p < 0.001). Conclusion The coexistence of NAFLD and hypothyroidism is more strongly associated with increased asprosin than is the presence of hypothyroidism alone. This could be due to increased insulin resistance as a possible link.