The level of IL-6 expression was determined in adipose cells by real-time quantitative PCR. To achieve this, the total RNA was extracted according to the kit protocol using an RNA extraction kit (Total RNA purification kit, Yekta Tajhiz Azma, Iran). The purity of extracted RNA was determined by calculating the 260/280 nm absorbance ratio, and the quality was evaluated by electrophoresis on agarose gel and observing the 28s and 18s ribosomal RNA bands. Using a cDNA Synthesis Kit (Yekta Tajhiz Azma), complementary DNAs (cDNA) were synthesized according to the kit's protocol and used for further investigation. Real-time PCR was performed using a real-time PCR kit (SYBER Green qPCR MasterMix 2×, Yekta Tajhiz Azma). PCR was performed at initial denaturation and enzyme activation (94 °C for 3 min), denaturation (95 °C for 5 s), annealing (62 °C for 10 s), and extension (72 °C for 15 s). After plotting the standard curves of the target (IL-6) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) reference genes, and calculating and comparing the performance of PCR, the 2−ΔΔCT equation was used to compare gene expression in different groups [22 (link)]. The primer sequences are presented in Table 1.