Opposite Effects of Two-Derived Antioxidants from Physalis pubescens L. on Hepatocellular Carcinoma Cell Line Malhavu
2016
Physalis pubescens L. (P. pubescens) is an edible plant used in folk medicine in China. There
is traditional, but not scientific, evidence for the anti-tumour effects of P. pubescens. This study aimed
to identify whether, or not, antioxidants rich in phenols and flavonoids from fruits and calyxes of P. pubescens
can be the candidates for further development of an anti-hepatoma fraction, and if such biological
effects coupled with reactive oxygen species (ROS) changes, can provide a direction for subsequent
biological action. The effects of calyx-origin (or fruit-origin) total phenol and flavonoid (CTPF or
FTPF) from P. pubescens on Malhavu cell viability were evaluated by using a counting-kit-8 (CCK-8)
method. Morphological characterisation of cells was undertaken and the structures were photographed
(200 × magnification) using Hoechst 3348 staining after exposure to different concentrations of CTPF
or FTPF. Induced-apoptosis activity was determined using flow cytometry (FC) after Annexin VFITC/
PI staining. The corresponding ROS changes in Malhavu cells were observed and quantified by
the uploading of 2’, 7’-dichlorofluorescin diacetate (DCFH-DA). Anti-oxidation was evaluated by a cellular
oxidation-stress model and chemical assessments for DPPH, hydroxyl radial, super-oxide radicals,
and reducing power. Result shows that CTPF led to significant anti-proliferation in a time- and dosedependent
manner. However, FTPF promoted cell viability at 100-1000 μg/mL with a dose-response
manner in 24 h. With the extension of exposure time to 48 h, the cell viability did not increase with the
growth of FTPF. Morphological characterisation and FC assay both demonstrated that CTPF, and not
FTPF possessed induced-apoptotic activity. CTPF potentially induced cell apoptosis by promoting oxidative
stress. FTPF indicated pro-oxidation at a concentration of 10 μg/mL and anti-oxidation capabilities
at higher concentrations. ROS scavenging assay by oxidation-stress model indicated that CTPF (10
- 400 μg/mL) had ROS inhibitory capacity (R 2 = 0.5156, p 2 = 0.5951, p <
0.0001). CTPF is a potential candidate requiring further exploration for the development of antihepatoma
ingredients. The down-regulation of cell viability was related to production and reduction of
cellular ROS.
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