Anti-senescence Activity of Indonesian Black Pepper Essential Oil (Piper nigrum L.) on ovarian CHO-K1 and fibroblast NIH-3T3 Cells

Background: Cellular senescence or cell aging is a progressive process caused by an increase in free radicals so that cells cannot proliferate. Black pepper essential oil (BPEO) contains terpenoid compounds that perform the antioxidant activity to inhibit cell aging. Aims: This study aims to explore the potential of BPEO as an anti-senescence agent to maintain body fitness through in vitro methods on ovarian CHO-K1 and fibroblast NIH-3T3 cells. Methods: BPEO was obtained by hydrodistillation, then identified the compounds by Gas Chromatography-Mass Spectrometry (GC-MS). The cytotoxicity of BPEO was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) Assay. The senescence was detected by senescence associated beta galactosidase (SA-β-Gal) assay. Also, the Reactive Oxygen Level (ROS) level effect was analyzed under 2',7'-dichlorofluorescein diacetate (DCFDA) staining assay with flow cytometry. Results: The BPEO contains 46 compounds, which the highest concentration compound is β-Caryophyllene, as much as 22,60 %. Cytotoxic tests indicated that BPEO up to the concentration of 200 µg/mL is not cytotoxic on CHO-K1 and NIH-3T3 cells. Furthermore, the administration of BPEO at concentrations of 50 µg/mL and 100 µg/mL reduces the number of senescent positive cells. This occurs seemingly irrespective of the ROS pathway. Conclusions: Overall, this study shows that BPEO contains active compounds that are safe for normal cells and has anti-senescence activities on CHO-K1 and NIH-3T3 cells. :TJPS-2020-0224.R1