Background: Designing the primer pairs is one of the most important factors in the amplification and quantitative analysis of the nucleic acid sequences of interest. Using in silico methods, the present study intends to design highly specific primers for quantitative analysis of the genes with minimum expression. To achieve this aim, we selected two candidate genes with little expression, namely, G-protein coupled receptor 120 (GPR120) and peroxisome proliferator-activated receptor-γ (PPARγ), in peripheral blood leukocytes of healthy volunteers. Methods: Peripheral blood was collected from 30 healthy volunteers. Primers for GPR120 and PPARγ were designed using online websites (UCSC, OligoCalc, and OligoAnalyzer) and the primer designing tool (NCBI). Total RNA extraction and cDNA synthesis were done using commercially available kits based on manufacturer instructions. Finally, the melting curve analysis of GPR120 and PPARγ was assessed using the quantitative real-time PCR method. Results: The in silico gene expression investigation revealed that GPR120 and PPARγ have minimal leukocyte expression. Besides, the melting curves analysis for both genes in the studied individuals showed only one melting peak, confirming the specific amplification of the desired genes. Conclusion: Altogether, the study findings indicated that we could utilize the peripheral blood sample for assessing the gene expression and amplification of omega-3 fatty acids receptors, i. e. GPR120 and PPARγ as two candidate genes with very low expression in leukocytes.

The reverse cholesterol transfer process has a positive role in reducing atherosclerotic plaques during diabetes. Therefore, the aim of the present study was to evaluate the effect of eight weeks of high-intensity interval training on GPR120 and AMPK proteins in the cardiac tissue of mice with type 2 diabetes. In an experimental study, 40 three-month-old adult male Wistar rats with an average weight (250-300 g) were randomly divided into four groups of 10 series including: healthy control (C: intraperitoneal injection of saline), healthy training (T: running at 85-90 % of maximum speed in 6 to 12 bouts in two-minute periods,5 days per week for eight weeks), diabetic control (D: diabetic on a high-fat diet with intraperitoneal injection of streptozotocin) and trained diabetics (D+T: diabetic with training) were divided. A method based on Western blotting was used to determine changes in the expression profile of GPR120 and AMPK proteins in the heart muscle tissue (left ventricle) of rats. The two-way analysis of variance and Bonferroni post hoc test were used to analyze the data. Induction of diabetes (D) significantly reduced GPR120 and AMPK proteins (p ≤ 0. 05). While HIIT training apply in healthy group (T) increased 81% and 47% in GPR120 and AMPK compared to diabetic control group (D) respectively (p = 0. 001). Also, training intervention has the ability to prevent a decrease in both of proteins in the trained diabetic group (D+T) compared to the diabetic group (D) (p = 0. 023). Based on the findings of this study, it can be stated that eight weeks of high-intensity interval training (HIIT) is a preventive strategy against reducing the activity of GPR120 and AMPK proteins involved in the reverse cholesterol transmission during type 2 diabetes.