Abstract
Purpose: To determine total phenolic and flavonoid contents, as well as the cytotoxic, immunemodulatoryand anti-inflammatory potentials of the whole plant of Astragalus creticus (Fabaceae).Methods: Folin-Ciocalteu (FCR) method was used for determination of total phenolic and flavonoid contents of the methanol and dichloromethane extracts of Astragalus creticus. The cytotoxic potential of the extracts on 3T3 and HeLa cell lines were evaluated using MTT assay. Brine shrimp larvae mortality was determined by lethality bioassay, while inhibitory effects were determined on mouse fibroblast (3T3)and cervical cancer (HeLa) cell lines. In vitro immunomodulatory and in vivo anti-inflammatory effectswere assessed using reactive oxygen species (ROS) chemiluminescence and formalin-induced rat paw edema assays, respectively.Results: Dichloromethane extract had higher contents of phenolics (TPC = 324.75 ± 2.47 mg GAE/g) and flavonoids (TFC = 95.51 ± 0.82 QE/g) than the methanol extract (TPC = 79.82 ± 1.53 mg GAE/g, TFC = 56.11 ± 0.93 QE/g). The dichloromethane extract exhibited high cytotoxic andimmunomodulatory potentials, with 76.66 % mortality in brine shrimp lethality bioassay and 83.9 % inhibition (IC50 = 18.0 ± 1.1 μg/mL) in chemiluminescence assay. The extract also resulted in 22 and 13 % inhibition of viability of HeLa and 3T3 cells, respectively, while the methanol extract produced 13 % inhibition of both cell lines. The methanol extract produced very significant anti-inflammatory activity,with a maximum of 49 % inhibition of paw edema at a dose of 160 mg/kg (p < 0.01).Conclusion: These results suggest that the dichloromethane and methanol extracts of Astragalus creticus (Fabaceae) exert cytotoxic, immunomodulatory and anti-inflammatory effects. These findings provide scientific validation for the traditional medicinal use of the Astragalus genus.
Highlights
Fabaceae is popularly known as the pea, legume or bean family which comprises 751 genera and 19000 species [1]
Astragalus creticus is an important plant of the Astragalus genus which is well known in the field of herbal medicine
The results showed that ACWPD produced better cytotoxic effect, with 6.66, 10 and 76.66 % mortality values, than ACWPM which produced mortality values of 10, 16.66 and 20.00 % against brine shrimp larvae at concentrations of 10, 100 and 1000 μg/mL, respectively
Summary
Fabaceae is popularly known as the pea, legume or bean family which comprises 751 genera and 19000 species [1]. Total phenolic contents (TPCs) were calculated from a gallic acid (GA) calibration curve, and expressed as mg of GA equivalents per gram of dry extract (mg GAE/g) [13]. The plant was collected near Fort Manro (tribal area of southern Punjab) in March, 2014 It was identified as Astragalus creticus by a taxonomist at Institute of Pure and Applied Biology, Bahauddin Zakariya University, Multan, and Determination of total flavonoid content (TFC). Plant extract (20 mg) was dissolved in 2 mL of solvent, and the sample was transferred at volumes of 5, 50, 500 μL to vials to generate concentrations of 10, 100, 1000 μg/mL, respectively. The absorbance of the formazan solution in each well was measured at 540 nm in a Spectra Max plus microplate reader, and the inhibition of cell viability (H)was calculated as shown in Eq 1. Soft-Max Proof software (US) was used for calculation [15]
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