In-vitro analysis of free radical scavenging activities and suppression of LPS-induced ROS production in macrophage cells by Solanum sisymbriifolium extracts

Abstract

The current study aims to evaluate the antioxidant, cytotoxicity activities and suppression of LPSinduced oxidative stress production and characterization of phytochemicals in Solanum sisymbriifolium leaf extracts. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity of the leaves of S. sisymbriifolium extracted with solvents of various polarities viz. water: ethanol, ratio 50: 50; ethyl acetate and dichloromethane, was assessed. The cytotoxicity of the extracts was determined using the [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide] (MTT) assay on RAW 264.7 macrophage (Murine) cells and real-time cell analysis (RTCA) xCELLigence system was used for determining cell viability. Cell-based detection of reactive oxygen species (ROS) was investigated utilizing a 2′,7′-Dichlorodihydrofluorescein diacetate (H 2 DCF-DA) assay. The DPPH and ABTS scavenging activity results of extracts revealed a dosedependent response with significantly lower activity in both DPPH and ABTS. The superoxide dismutase (SOD) enzyme activity was then evaluated and extracts displayed a high SOD enzyme activity with 90-50% activity. Cytotoxicity results revealed that S. sisymbriifolium extracts were not toxic to RAW 264.7 macrophage cells at the tested concentrations. All three extracts decreased the production of ROS in macrophage cells. Phytochemical analysis using Fourier-transform infrared spectroscopy (FTIR) indicated the presence of metabolite functional groups which may be responsible for the antioxidant activity. the current study indicates that S. sisymbriifolium contains phytochemicals that scavenge free radicals, with less toxicity, and suppresses the LPS-induced ROS production in RAW 264.7 macrophage cells.

Chemical Composition, In Vitro and In Silico Antioxidant Potential of Melissa officinalis subsp. officinalis Essential Oil

“…Their results clearly showed that the most active tested extracts and fractions inhibited DPPH • and ROS formations in a dose-dependent manner. A more recent study evaluated the antioxidant activity of several Solanum sisymbriifolium extracts showing that the tested products also demonstrate the ability to scavenge DPPH • and ABTS • and, at the same time, reduce ROS production in a dose-dependent manner [62]. Although these correlations may be difficult to consider due to the complex composition of a plant extract, they are also reported in cases when the antioxidant activity of a single chemical compound is determined.…”

Application of xCELLigence real-time cell analysis to the microplate assay for pertussis toxin induced clustering in CHO cells

“…https://doi.org/10.1371/journal.pone.0248491.t004 [39][40][41][42][43][44][45][46][47][48][49][50][51][52][53][54][55][56][57]. Although the xCELLigence system has been previously used to monitor cell interaction with other bacterial toxins such as Vibrio cholerae toxin [58] and Clostridium difficile toxin [59][60][61][62], to our knowledge, this is the first demonstration of the system suitability to detect PT.…”

Anti‑inflammatory effects of Fritillaria thunbergii Miquel extracts in LPS‑stimulated murine macrophage RAW 264.7 cells

“…ABTS is a relatively stable free radical and is widely used for measuring antioxidant activity along with DPPH. This method can measure both lipophilic or hypophilic substances ( 30 ). In this study, EAFM improved the ABTS radical scavenging ability.…”