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Background Obesity and diabetes are two chronic metabolic diseases whose prevalence is increasing at an alarming rate globally. A close association between obesity, diabetes, and insulin resistance has been identified, and many studies have pinpointed obesity as a causal risk factor for insulin resistance. However, the mechanism underlying this association is not entirely understood. In the past decade, ceramides have gained attention due to their accumulation in certain tissues and their suggested role in initiating insulin resistance. This study aims to determine the association of specific ceramides and their major metabolizing enzymes with obesity-associated insulin resistance. Methods The samples comprised subcutaneous adipose tissues collected from three cohorts: lean non-diabetic (controls;  n  = 20), obese-non-diabetic ( n  = 66), and obese-diabetic ( n  = 32). Ceramide levels were quantified using LC-MS/MS and mRNA expression level for different enzymes were estimated using real-time PCR-based RNA expression analysis. Results C16-ceramide ( P  = 0.023), C16-dihydro-ceramide ( P  &lt; 0.005), C18-dihydro-ceramide ( P  = 0.009) and C24-ceramide ( P  = 0.040) levels were significantly increased in the obese cohort compared to the control group. However, stratification of the obese group revealed a significant increase in the C16-ceramide levels ( P  = 0.027) and mRNA over expression of the serine palmitoyl transferases enzyme subunit SPT1 ( P  &lt; 0.005) in the obese-diabetic cohort compared to the obese-non-diabetic cohort. Conclusions The present study indicates that C16-ceramide plays a pivotal role in inducing insulin resistance. Overexpression of  SPT1  in the obese-diabetic group and its positive correlation with C16-ceramide suggest that C16-ceramide was generated through the  de novo  pathway.

Insulin resistance induced by de novo pathway–generated C16-ceramide is associated with type 2 diabetes in an obese population