Myocardial tissue is sensitive to insulin resistance (IR) due to its interactions with insulin levels. Previous studies demonstrated that heart failure prevalence was higher in IR patients. Evaluation of myocardial deformation by multilayer global longitudinal strain (MGLS) might provide more information about IR related left ventricular dysfunction. In this study, we aimed to investigate subclinical LV dysfunction with MGLS in patients with IR. The study was designed as a prospective cross-sectional study. The present study included 64 patients with IR (+), and 54 subjects without IR (-) prospectively. The homeostasis model of insulin resistance (HOMA-IR) was used to quantify insulin resistance. Specific analysis for endocardial, mid-myocardial and epicardial layers were performed by two-dimensional (2D) speckle tracking echocardiography (STE) for multilayer global longitudinal strain. MGLS (Endocard-Mid-myocard-Epicard) values were significantly lower in the IR (+) group compared to IR (-) group ((GLS-endocard; - 15.1 +/- 1.5 vs. - 18.7 +/- 1.3, p < 0.001), (GLS-mid-myocard; - 16.0 +/- 2.0 vs. - 18.0 +/- 2.0, p < 0.001), (GLS-epicard; - 17.0 +/- 1.7 vs. - 18.01 +/- 1.94, p = 0.004)). GLS-endocard levels were significantly and positively correlated with HOMA-IR levels (r = 0.643, p < 0.001). HOMA-IR and age were found to be independent factors in detecting a decrease in GLS-endocard level in regression analysis. In conclusion, our data reveal that IR (+) patients had significantly lower strain values compared to IR (-) group. Besides, we presented that the HOMA-IR value was an independent predictor of subclinical left ventricular dysfunction.