Effect of cinnamon water extract on monocyte-to-macrophage differentiation and scavenger receptor activity

Kang, Hee; Park, Sung-Hyun; Yun, Jeong-Moon; Nam, Tae-Gyu; Kim, Young-Eun; Kim, Dae-Ok; Kim, Youn Jung

THP-1 cells (2 × 10 6 cells) were cultured with PMA (100 nM) in the presence of CWE for 48 h. Total RNA was extracted using an RNeasy mini kit (Qiagen, Germany) according to the manufacturer's instructions, and revere transcribed using Superscript III reverse transcriptase (Invitrogen, CA, USA). The cDNA obtained was mixed with Power SYBR Green PCR Master mix (Applied Biosystems, CA, USA) and 2 pmol primers in a final volume of 20 μl. The following forward and reverse primer sequences were used: SRA, forward: 5′-GCA GTG GGA TCA CTT TCA CAA-3′ and reverse: 5′ AGC TGT CAT TGA GCG AGC ATC-3′; CD36, forward: 5′-GCC AAG GAA AAT GTA ACC CAG G −3′ and reverse: 5′-GCC TCT GTT CCA ACT GAT AGT GA-3′; and actin, forward: 5′-GCAAATGCTTCTAGG CGGACTAT-3′ and reverse: 5′-TGTTTTCTGCGC AAGTTAGGTTT-3′. PCR was performed in triplicate using a StepOne Real-time PCR system (Applied Biosystems). After an initial heat denaturation at 95°C for 10 min, the PCR conditions were set for 40 cycles at 95°C for 15 s and 60°C for 1 min. Relative gene expression was determined using the standard curve method and normalized to actin.