Ab142044

In order to evaluate the effect of cinnamon samples on the activity of HepG-2 and BJ-1, cells were seeded in RPMI-1640 medium containing 25 mg/mL of each cinnamon sample, with one 106 cell per flask, and incubated for 48 h at 37 °C under a humidified atmosphere of 5% CO₂. After this, the cell medium was converted to serum-free medium (SFM) comprising 10 µL/mL of yogurt extract. Then, trypsin 0.05%/0.53 mM EDTA solutions were used to trypsinize the cell cultures after incubation. After washing in PBS, the cells were centrifuged at 2000 rpm for 5 min at 4 °C and resuspended in 1 mL PBS containing 0.1% Triton X-100. Using a 1.5 mL micro centrifuge tube, cells were sonicated twice for 10 s at 100 Hz (Vibra-cell, Sonics & Material) and centrifuged for 30 min at 4 °C at 14,000 rpm. The cells were sonicated for 2 min at 100 Hz in a 1.5 mL micro centrifuge tube placed on ice, then centrifuged at 14,000 rpm for 30 min at 4 °C after the sonication (Vibra-cell, Sonics & Material). Enzyme activity was tested in the supernatant of the tube. We measured the enzyme activity of SOD, CAT, GSH, and GPx in cell culture according to the manufacturer’s instructions for colorimetric kits ab65354, ab83464, ab142044, and ab102530 Abcam.

Estimation of pancreatic tissue content of malondialdehyde (MDA) was done by using the thiobarbituric acid reaction explained by Draper et al., and the result was stated in nmol/g tissue [20 (link)]. Total antioxidants were analyzed according to the method described by Koracevic et al. via the kit (Cat. No. # TA 25 12) delivered from Bio-Diagnostics, Dokki, Giza, Egypt. The result was presented as mmol/L [21 (link)].
Catalase activity (Catalase Activity Assay Kit-ab83464, Abcam, Hong Kong, China) reduced glutathione (Reduced Glutathione (GSH), Endogenous antioxidant ab142044, Abcam) and Superoxide Dismutase (SOD) (Superoxide Dismutase Activity Assay Colorimetric Kit-ab65354, Abcam) were determined according to the manufacturer’s instruction in serum samples [22 ,23 ,24 (link)]. The parameters were represented as U/mL, μmol/mL, and U/mL, respectively.