The aims of this work were to study growth, isothiocyanate (ITC), bioactive content, antioxidant activity and anticancer activity of mustard green (MG) microgreens grown from seeds treated with cold plasma at 21 and 23 kV for 5 min. Microgreens from plasma-treated seeds at 23 kV showed almost 2-fold increased ITC content (1.57 ± 0.05 mmol/100 g DW) compared to MG from seeds without plasma (control), showed the highest total phenolic content (TPC) (6.76 ± 0.14 mg GAE/g DW) and total flavonoid content (TFC) (0.16±0.01 mg RE/g DW). However, MG plasma-treated seeds at 21 kV showed the highest antioxidant activity from 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assay (3.51 ± 0.38 mg TE/g DW). Allyl isothiocyanate and 3-butenyl isothiocyanate were the dominant ITCs in MG. The highest cytotoxicities using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay against MCF-7 (IC50 of 32.44 ± 1.64 μg/mL) and HepG2 (IC50 of 28.58 ± 1.04 μg/mL) after 72 h exposure were found in MG from plasma-treated seeds at 23 kV and MG from control seeds, respectively. However, MG from plasma-treated seeds at 21 kV exhibited the highest antiproliferative effect against MCF-7 (IC50 of 23.23 ± 0.23 μg/mL) and HepG2 (IC50 of 20.44 ± 0.56 μg/mL) for 14 days and also the most potent antimigratory effect. MG from cold plasma inhibited MMP-9 protein expression in both cancers indicating antimigratory property. MG from cold plasma also significantly reduced MMP-9 mRNA expression in both cancers when compared to the control and untreated cells. In conclusion, cold plasma treatment on seeds seemed to be an innovative tool to enhance ITC, TPC, TFC and anticancer properties of MG microgreens for better health implications. © 2021 Friends Science Publishers
BACKGROUND: Raphanus sativus var. caudatus or Thai rat-tailed radish (RTR) contains glucosinolates and isothiocyanates with chemopreventive effects; however, only mature plants have been investigated to date. Thus, the present study aimed to determine isothiocyanates, phenolic compounds and flavonoid compounds, antioxidant activity, cytotoxicity, and antiproliferative activity of RTR microgreens grown from seeds treated with cold plasma (21 kV for 5 min), organic elicitor (160 mmol L −1 NaCl, 10 mmol L −1 CaCl 2 or 176 mmol L −1 sucrose) or both in combination. Seeds were germinated on vermiculite and sprayed with deionized water or elicitor for 7 days before harvest. RESULTS: Cold plasma had insignificant effect on growth, whereas NaCl and CaCl 2 increased fresh weight. Plasma with CaCl 2 led to the highest total isothiocyanate (ITC) content [1.99 g kg −1 dry weight (DW)] in RTR microgreens containing raphasatin as the only ITC detected. Plasma treatment gave the highest total phenolic content (7.56 mg gallic acid equivalents g −1 DW), antioxidant activity from a 2,2-diphenyl-1-picrylhydrazyl assay (7.70 mg trolox equivalents g −1 DW) and ferric reducing antioxidant power assay (21.72 mg Fe 2+ g −1 DW). Microgreen extracts from plasma showed an IC 50 value of 29.28 and 13.83 ∼g mL −1 towards MCF-7 and HepG2, respectively, with inhibitory properties on matrix metalloproteinase (MMP)-2 and MMP-9 proteins. Plasma enhanced Bax and Caspase-3 gene expression but reduced Bcl-2 and MMP-9 expression, indicating activation of apoptosis. CONCLUSION: Cold plasma shows promise as an innovative tool to enhance bioactive compounds with chemopreventive benefits in microgreens.
This work aimed to evaluate the antioxidant capacities and cytotoxic attributes of Alternanthera sissoo (AS) and Alternanthera bettzickiana Green (AB) which are commonly grown in Thailand, yet not extensively-studied. Dried leaves were extracted by maceration using ethanol for 48 h and ethanolic extract solution was used. Cytotoxic, anti-proliferative, anti-migratory capacities and apoptosis-related gene expressions of AS and AB extracts on MCF-7, HepG2, and HeLa cell lines were investigated using 3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetra zolium bromide (MTT) assay, anti-colony formation and wound healing test, respectively. The antioxidant activities by DPPH assay and FRAP assay of AS (130.62±1.61 µg TE/g extract and 24.76 ± 0.54 mg Fe(II)/g extract, respectively) were significantly higher than those of AB (104.34±3.68 µg TE/g extract and 23.13± 0.90 mg Fe(II)/g extract, respectively). Likewise, total phenolic content and the total flavonoid content of AS (7.43±0.39 mg GAE/g and 184.22±3.20 mg RE/g extract, respectively) were significantly higher than AB (6.10±0.29 mg GAE/g extract and 168.07±7.90 mg RE/g extract, respectively). HPLC analysis showed the predominance of myricetin, rutin and ferulic acid. For MCF-7, HepG2 and HeLa, the lowest IC50 values of 48.53 µg/mL, 69.94 µg/mL and 43.98 µg/mL, respectively were found in AS extract at 72 h exposure using MTT assay. Apoptotic bodies, gene expression and protein expressions related to apoptosis were more pronounced in AS treatment. Both AS and AB can be served as local foods with chemopreventive effects against breast, liver and cervical cancers. This work provided a foundation for future pharmacological research on Alternanthera plant extracts.
Cervical cancer is second only to breast cancer in terms of incidence; however, it is the most lethal form of cancer among Thai women due to the asymptomatic nature of its early stages. This work aimed to examine cytotoxic and antiproliferative capacities of Rhinacanthus nasutus (RN) and Acanthus ebracteatus (AE) extracts against human cervical cancer cells (HeLa). Plant leaves were used for ethyl acetate extraction. The antioxidant assays, HPLC analysis, a cytotoxic MTT assay, a clonogenic assay and real-time PCR were conducted. Both RN and AE displayed similar DPPH scavenging activity (3.97 and 4.05 mg TE/g DW) and ferric reducing antioxidant power (4.79 and 4.35 mg Fe2+/g DW). However, AE was richer in total phenolic content than RN (13.30 and 10.84 mg GAE/g DW, respectively). Rutin, catechin, chlorogenic acid, and cinnamic acid were found in AE, whilst only cinnamic acid with much higher content was found in RN. Higher cytotoxicity of 91.73% against HeLa cells was found in RN (IC50 value of 62.06 µg/mL). RN showed higher antiproliferative effect (IC50 of 25.24 µg/mL) than AE (34.35 µg/mL). Genes (Bcl-2, Bax, MMP-2, MMP-9, caspase-3, p21, and cyclin D1) and proteins (cytochrome c, caspase-3 and p21) linked to apoptosis and migration were substantially more affected by RN. To conclude, both RN and AE hold promise as anticancer herbal plants against human cervical cancer; however, RN was more cytotoxic and antiproliferative in HeLa cells. RN offers a better alternative herbal medicine or complementary remedy to the standard drug for human cervical cancer treatment.
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