Nutr Res Pract.  2012 Oct;6(5):389-395.

Carbohydrase inhibition and anti-cancerous and free radical scavenging properties along with DNA and protein protection ability of methanolic root extracts of Rumex crispus

• Shiwani S
• Singh NK
• Wang MH

Abstract

The study elucidated carbohydrase inhibition, anti-cancerous, free radical scavenging properties and also investigated the DNA and protein protection abilities of methanolic root extract of Rumex crispus (RERC). For this purpose, pulverized roots of Rumex crispus was extracted in methanol (80% and absolute conc.) for 3 hrs for 60degrees C and filtered and evaporated with vacuum rotary evaporator. RERC showed high phenolic content (211 microg/GAE equivalent) and strong 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging (IC50 = 42.86 (absolute methanol) and 36.91 microg/mL (80% methanolic extract)) and reduced power ability. Furthermore, RERC exhibited significant protective ability in H2O2/Fe3+/ascorbic acid-induced protein or DNA damage and percentage inhibition of the HT-29 cell growth rate following 80% methanolic RERC exposure at 400 microg/mL was observed to be highest (10.2% +/- 1.03). Moreover, methanolic RERC inhibited alpha-glucosidase and amylase effectively and significantly (P < 0.05). Conclusively, RERC could be considered as potent carbohydrase inhibitor, anti-cancerous and anti-oxidant.

Keyword

Root of Rumex crispus; methanolic extract; antioxidative; carbohydrase inhibitor; anti-cancerous

MeSH Terms

alpha-Glucosidases
Amylases
Biphenyl Compounds
DNA
DNA Damage
Glycoside Hydrolases
HT29 Cells
Humans
Methanol
Phenol
Picrates
Power (Psychology)
Rumex
Vacuum
Amylases
Biphenyl Compounds
DNA
Glycoside Hydrolases
Methanol
Phenol
Picrates
alpha-Glucosidases

Figure

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  Fig. 1 Reducing power potential of absolute and 80% methanolic extracts of Rumex crispus roots. α-Tocopherol and BHT were used as positive controls. Each value was expressed as mean ± SD (n = 3). *(P < 0.05) representing significant values.

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  Fig. 2 Metal chelating activity (A) and superoxide radical scavenging activity (B) of absolute and 80% methanolic extracts of Rumex crispus roots. EDTA and galic acid was used as positive control. Each value was expressed as mean ± SD (n = 3). *P < 0.05 and **P < 0.01 representing significant values.

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  Fig. 3 Cell apoptosis representation of human cancer cells (HT-29) under the influence of absolute and 80% methanolic extracts of Rumex crispus roots. (A): Phase contrast microscopic examination of cell apoptosis of human cancer cells (HT-29); 1) Control; 2) 80% methanolic extract; 3) Absolute methanolic extract. (B): Cell apoptosis % of human cancer cells (HT-29) at different concentration of root extracts of Rumex crispus in culture for 48 h. Cell viability was measured using an MTT assay. Data represent the mean ± SD of three independent experiments.

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  Fig. 4 Influence of absolute and 80% methanolic extracts of Rumex crispus roots. A: α-glucosidase (at 200 µg/mL). B: α-amylase (at 400 µg/mL concentration). Each value was expressed as mean ± SD (n = 3). *P < 0.05 and **P < 0.01 representing significant values.

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  Fig. 5 Protection of BSA oxidative damage by absolute and 80% methanolic extracts of Rumex crispus. The electrophoretic pattern of BSA and densitometric analysis of the corresponding band intensity has been depicted in all the graphs representing different concentration of the extracted samples i.e. A (100 µg/mL), B (500 µg/mL), C (1,000 µg/mL) and D (5,000 µg/mL). BSA was oxidised by Fenton system (Fe3+/H2O2/ascorbic acid). Each bar represents the mean ± SD of three different experiments. Means with different letters at a time differ significantly, P < 0.05. The values sharing common letters are not significantly different at P < 0.05. *P < 0.05 compared with Fe3+/H2O2/ascorbic acid treated group.

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  Fig. 6 HEK 293 cellular DNA protection from hydroxyl radical with methanolic (80% and absolute) extract from Rumex crispus has been shown on agarose gel electrophoresis and quantification of DNA band has been indicated on bar graphs. Histogram showing the protective effect of different concentration of extract i.e. A (100 ug/mL), B (250 ug/mL), C (500 ug/mL) and D (1,000 ug/mL) against DNA damage based on densitometric measurements. Line 1, DNA incubated without Fenton's reagent; Line 2-3, DNA incubated with Fenton's reagent in the presence of 100, 250, 500 and 1,000 µg/mL, respectively; line 4: DNA incubated with Fenton's reagent.

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