Radical scavenging capacities and inhibition of human prostate (LNCaP) cell proliferation by Fortunella margarita

Abstract

Lyophilised nagami kumquat (Fortunella margarita) powder was extracted with five different solvents. Dried extracts of EtOAc and MeOH–water (4:1, v/v) had the highest and lowest total phenolics, respectively, by Folin–Ciocalteu method. LC–MS analysis confirmed the presence of different levels of rutin, narirutin, poncirin, apigenin 8-C-rutinoside and 3′,5′,di-C-β-glucopyranosyl phloretin in all the extracts except n-hexane. EtOAc and MeOH extracts exhibited the highest and the lowest 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, respectively. The order of antioxidant capacity was found to be MeOH–water (4:1, v/v)>EtOAc>MeOH>n-hexane>acetone by phosphomolybdenum complex and oxygen radical absorbance capacity (ORAC) values. Among five extracts, n-hexane extract exhibited the highest inhibition of human prostate cancer (LNCaP) cells (86.4%) after 96h at 100μg/mL, followed by EtOAc (82.8%), MeOH (76.7%) and MeOH–water (4:1, v/v) (68.2%). Fragmentation of DNA suggests the ability of extracts to induce apoptosis in LNCaP cells. The cleavage of caspase-3 was the highest in n-hexane and EtOAc extracts, whereas the ratio of Bax/Bcl2 was the highest in MeOH and MeOH–water (4:1, v/v) extracts. The results of the present study were also supported by fluorescent images of LNCaP cells treated with kumquat extracts. The maximum cell proliferation inhibition activity of n-hexane extract may be due to the presence of one or cumulative effect of β-carotene, β-cubebene and hexadecanoic acid. Remaining four extracts exhibited differential antioxidant activity and cytotoxicity which may be due to the presence of various levels of rutin, narirutin, poncirin, apigenin-8-C-rutinoside and 3′,4′-di-C-β-glucopyranosyl phloretin in each extract.