Basic HTML Version
Medicinal Plant Research 2014, Vol.4, No.4, 30
-
34
http://mpr.sophiapublisher.com
33
measurement of samples was carried out with three
replicates. The percentage of radical scavenging
activity was calculated according to the following
equation (Liu et al., 2008):
where
is the absorbance of DPPH reagent with
80% methanol as negative control,
is the
absorbance of reaction mixture and
is the
absorbance of sample which is not containing DPPH
solution and this term is used for error correction
because of the natural colour of the sample.IC
50
value
was calculated by using correlation and regression
analysis in the Graph Pad Prism (San Diego, USA).
3.3 Cell culture preparation
Six cancer cell lines and one non-cancerous cell line
were used for determination of cytotoxic activity.
Human colon adenocarcinoma cell line (HT
-
29),
human hepatoma cell line (Hep3B), human breast
adenocarcinoma cell line (MCF
-
7), human neurobla-
stoma cell line (SH-SY5Y), human osteosarcoma cell
line (SaOS
-
2), human prostate adenocarcinoma
(LNCap) cell line were selected as cancer cell lines.
Kidney epithelial cells from an African green monkey
cells (Vero) was also used as non-cancerous cell line.
All cell lines except LNCap were maintained in
Dulbecco’s modified Eagle’s medium (DMEM)
(Biochrom AG, Germany), supplemented with 10%
fetal bovine serum (FBS) (Biocrom AG, Germany),
2 mM L-glutamine, 1 g/L glucose, 100 U/mL of
penicillin and 100 µg/mL of streptomycin (Biochrom
AG, Germany). LNCap cells were maintained in
RPMI 1640 (Biochrom AG, Germany) medium
supplemented with 10 % FBS, 2mM L-glutamine,
100 U/mL of penicillin and 100 µg/mL of streptomy-
cin (Biochrom AG, Germany). The cultures were
incubated at 37°C in a humidified atmosphere of 5%
CO
2
(Heraus, Germany).
3.4 Cytotoxicity assay
The percentage of apoptotic cells was calorimetrically
measured using MTT, according to the manufacturer’s
instructions (Sigma-Aldrich, USA). Cells were
incubated at densities of 1×10
5
cells/well in 96-well
plates containing 100 µL of DMEM medium (RPMI
medium for LNCap cells) overnight. Cultures were
treated with extract at different concentrations
(1 µg/mL, 5 µg/mL, 10 µg/mL, 30 µg/mL, 50 µg/mL)
for 24 and 48 hours. After incubation period, culture
medium was changed with FBS-free medium
containing MTT (5 µg/mL) and incubated for 4 hours
at 37°C. Medium was discarded and consisting
formazan crystals were solubilized with DMSO
(dimethyl sulfoxide). Optical densities were measured
at 570 and 690 nm with a multiwell spectropho-
tometer (VersaMax, USA).
Authors’ contributions
All the authors contributed equally for this study. All the
authors read and approved the final version of the manuscript.
Acknowledgement
The authors would like to thank Professor S. İsmet Deliloğlu
Gürhan (Department of Bioengineering, Faculty of Engineering,
Ege University, Izmir, Turkey) for her valuable comments and
suggestions during the experimental studies.
References
Lan Y.H., Chang F.R., Pan M.J., Wu C.C., Wu S.J., Chen S.L., Wang S.S.,
Wu M.J., and Wu Y.C., 2009, New cytotoxic withanolides from
Physalis peruviana,
Food Chem., 116: 462-469
Puente L.A., Pinto-Muñoz C.A., Castro E.S., and Cortés M., 2011,
Physalis
peruviana
Linnaeus, multiple properties of a highly functional fruit: A
review, Food Res. Int., 44: 1733-1740
Ramadan M.F., 2011, Bioactive phytochemicals, nutritional value and
functional properties of cape gooseberry (
Physalis peruviana
): An
overview, Food Res.
Int. 44: 1830- 1836
Ramadan M.F., and Moersel J.T., 2007, Impact of enzymatic treatment on
chemical composition, physicochemical properties and radical
scavenging activity of goldenberrry (
Physalis peruviana
L.) juice, J.
Sci. Food Agr., 87: 452-460
Reddy C.V.K., Sreeramulu D., and Raghunath M., 2010, Antioxidant
activity of fresh and dry fruits commonly consumed in India, Food Res.
Int. 43: 285-288
Rodriguez A.T., Moreno Y.S., Guadarrama S.V., and Tejacal I.A., 2011,
Soluble phenols and antioxidant activity in mameysapote (
Pouteria
sapota
) fruits in postharvest, Food Res. Int., 44: 1956- 1961
Salazar M.R., Jones J.W., Chaves B., and Cooman A., 2008, A model for the
potential production and dry matter distribution of Cape gooseberry
(
Physalis peruviana
L.), Sci. Hortic., 115: 142-148
Vasco C., Ruales J., and Kamal-Eldin A., 2008, Total phenolic compounds
and antioxidant capacities of major fruits from Ecuador, Food Chem.
111: 816-823
Wu S.J., Chang S.P., Lin D.L., Wang S.S., Hou F.F., and Ng L.T., 2009,
Supercritical carbon dioxide extract of
Physalis peruviana
induced cell