Cafestol

Cell lines

Human renal carcinoma cells (Caki), human breast carcinoma cells (MDA-MB231), human glioma cells (U251MG), human colon carcinoma cells (HCT116), human leukemia cells (U937), and human prostate carcinoma (PC3) cells

Preparation Method

Human renal carcinoma cells (Caki), human breast carcinoma cells (MDA-MB231), human glioma cells (U251MG), human colon carcinoma cells (HCT116), human leukemia cells (U937), and human prostate carcinoma (PC3) cells were obtained from the American Type Culture Collection. All cells were cultured in Dulbecco’s Modified Eagle’s Medium containing 10% fetal bovine serum, 20mM HEPES buffer, 100U/ml penicillin, 100μg/ml streptomycin, and 100μg/ml gentamicin. A cell death detection ELISA plus kit was used for assessing apoptotic activity by detecting fragmented DNA within the nucleus in ABT-737-, cafestol-, and combination of ABT-737 and cafestol-treated cells. Briefly, cells were treated with different concentrations of cafestol(20 or 30μM) and ABT-737 alone or in combination. After 24h, each culture plate was centrifuged for 10min at 200×g, the supernatant was removed, and the pellet was lysed for 30min. After centrifuging the plate again at 200×g for 10min, the supernatant that contained the cytoplasmic histoneassociated DNA fragments was collected and incubated with an immobilized anti-histone antibody. The reaction products were incubated with a peroxidase substrate for 5min and measured by spectrophotometry at 405 and 490nm (reference wavelength) with a microplate reader. The signals in the wells containing the substrate alone were subtracted as the background.

Reaction Conditions

20 or 30μM; 24h

Applications

Cafestol significantly potentiated ABT-737-induced apoptosis.

Animal models

Wistar albino experimental rats

Preparation Method

A total of 32 Wistar albino experimental rats of both sexes aged 6 weeks with an average weight of 150g were used in this study; animals were acquired and kept under controlled conditions of a light/dark cycle (12h) at a temperature of 23°C±2°C and humidity of 50% ± 5%. Experimental animals were randomly assigned into four groups (n = 8) as follows: Group I: each rat was given vehicle only (5% Tween in DDW) orally via oral gavage for 14 consecutive days. Then, a single dose of NaCl (0.9%), 10mL/kg, was injected intraperitoneally 1h after the last vehicle administration on day 14. This group served as the normal (negative control) group. Group II: each rat was orally given cafestol (5mg/kg/day) for 14 consecutive days. Group III: each rat was given vehicle only (5% Tween in DDW) via oral gavage for 14 consecutive days. Then, a single dose of doxorubicin (15mg/kg) was injected intraperitoneally 1h after the last vehicle administration on day 14 to serve as the positive control group. Group IV: each rat received cafestol (5mg/kg/day) orally for 14 consecutive days, and then, a single dose of doxorubicin (15mg/kg) was injected intraperitoneally 1h after the last cafestol treatment on day 14. 24 hours after doxorubicin dose administration (i.e., day 15), the animals were anaesthetized and samples were collected for further analysis.

Dosage form

5mg/kg/day for 14 days; p.o.

Applications

Cafestol significantly protected against doxorubicin-induced cardiotoxicity by attenuating oxidative stress, activating the Nrf2/HO-1/NQO-1 pathway, suppressing inflammation, and improving cardiac biomarkers and histopathology.

References:
[1] Woo, S. M., Min, K. J., Seo, B. R., Nam, J. O., Choi, K. S., Yoo, Y. H., & Kwon, T. K. (2014). Cafestol overcomes ABT-737 resistance in Mcl-1-overexpressed renal carcinoma Caki cells through downregulation of Mcl-1 expression and upregulation of Bim expression. Cell death & disease, 5(11), e1514.
[2] Al-Kenany, S. A., & Al-Shawi, N. N. (2023). Protective effect of cafestol against doxorubicin-induced cardiotoxicity in rats by activating the Nrf2