(食品质量与安全研讨会)In vitro effects of anti-oxidants from

Anti-oxidative CTPF is a well bioactive compound deserving further separation to the development of anti-hepatoma ingredients.
FTPF improved viability of hepatoma cell line within tested concentrations, which suggests that anti-oxidants may not always be a beneficial alternative for a high-risk cancer population.
diluted with complete medium containing 10% FBS under aseptic conditions.
dissolved in DMSO solvents, filtrated with 0.22μm aseptic Millipore filter
CTPF CTPF
My presentation will cover the following aspects:
Preparation of tested compounds Determination of flavonoid and phenolic content Cell culture Cell proliferation assay Induced-apoptotic property assay Measurement of ROS in Malhavu cells exposed
However within the tested
concentrations,
FTPF
presented an obvious
promotion in the cell
viability.
Cell viability (%)
Cell viability (%)
140
A
120 100
80 60 40 20
C
A: treated cells with CTPF B: treated cells with FTPF
Incidence expressed as the sum of early and late apoptotic incidence.
Fig. 4. A1, B1, C1. Oxidation-stress effects on Malhavu cells——DCFH-DA
Cell culture——Hepatocellular carcinoma
cell line Malhavu（ Mal. ）
DMEM medium 10% FBS 2 mM glutamine in a humidified atmosphere 5% CO2 ；37 °C
propagated every 3 to 5 days
Preparation of tested compounds
stored at -20 °C
until use
Bing dried
in oven at
50 °C for
24 h
Powdered,
stored at -20
°C
80% ethanol, at 60 °C, 3 h, ratio of sample-tosolvent : 1:70 (w/v)
Table 1 Determination of phenols and
flavonoids in TPF from P. pubescens
Flavonoids (mg of gallic acid equivalent /g)
Polyphenols (mg of quercetin
equivalent /g）
Additionally, the cellular assessment is appropriate to reveal anti-oxidation of different compounds due to the close of physiological dose.
Conclusions
Calyxes origin 78.16± 9.92
102.45± 7.98
Fruits origin 30.30± 0.47
45.00± 1.67
Total flavonoid content：color rendering method of aluminium
nitrate Total phenolic content ：Folin-Ciocalteu method
Fig. 5. A, B, C, D Anti-oxidative assessment of CTFP or FTPF against different free radicals.
Discussion
Dual action of CTPF in different condition
Failure to anti-hepatoma effect of antioxidative FTPF——antioxidant may be inefficient for patients with cancer
0 50
140
B
120 100
80 60 40 20
0 50
100 200 400 600 800 Concentration of TPF (μg/mL) (24h)
1000
100
200

400
600
800
Concentration of TPF (μg/mL) (48h )
1000
Control DMSO control Treated groups (FTPF) Treated groups (CTPF)
Control DMSO control Treatment groups ( FTPF ) Treatment groups ( CTPF )
Fig. 2. Malhavu cells dyed by Hoechst 33348.
A: treated cells with FTPF. B: treated cells with CTPF. A1 and B1: cells are photographed under normal pattern. A2 and B2: cells are photographed under fluorescence pattern.
objective
This study aimed to identify whether, or not, antioxidants rich in phenols and flavonoids from fruits and calyxes of P. pubescens can be a candidate for the further development of an antihepatoma fraction, and if such biological effects coupled with reactive oxygen species (ROS) changes, can provide a direction for subsequent biological assay.
200
600 0.2%DMSO
100
200
Concentration of TPFs (μg/mL)
Early apoptotic incidence (%) for FTPF Late apoptotic incidence (%) for FTPF Early apoptotic incidence (%) for CTPF Late apoptotic incidence (%) for CTPF
Thanks for your attentions!
(食品质量与安全研讨会)In vitro effects of anti-oxidants from fruits and calyxes of Physalis pubescens L. on hepatocellular carcinoma cell line Malhavu
Laboratory studies have shown that various therapeutic effects are generally involved in plants of Physalis such as: antibacterial, anti-inflammatory, anticancer, immune adjustment and regulation of blood lipid level effects.
Fig. 3. Cell apoptosis determined by Annexin V-FITC/PI staining
A
40
NS
NS
30
NS
NS
B
40
** ***
30
Apoptotic incidence(%) Apoptotic incidence(%)
20
20
10
10
0
0
0.6%DMSO
Fig. 1. Effects of TPF from P. pubescens on
cell viability.
Fig. 1. shows that the two
tested
compounds
produced
different
biological effects on Mal
cells.
CTPF led to a significant dose and time-depend anti-proliferative property.