GC-MS analysis, anti-inflammatory and anti-proliferative properties of the aerial parts of three Mesembryanthemum spp

Abstract

Background: Due to their variability and safety, widespread research on phytochemicals continually encourages researchers to study various plants for their potential health benefits.

Objectives: This study aims to explore the phytochemical constituents of the aerial parts of three Mesembryanthemum spp.; M. nodiflorum, M. forsskaolii, and M. cordifolium existed in Egyptian nature using GC-MS analysis and studying their different biological activities in correlation to computational analysis.

Methods: Investigation of in vitro anti-inflammatory and anticancer activities and in silico studies of identified major compounds on VEGFR. Results: Thirty-three compounds were identified, octadecanoic acid, 2, 3-dihydroxypropyl ester, and 1H-Indene, 1-hexadecyl-2, 3-dihydro are the common compounds in the three extracts with different percentages. M. forsskaolii is the most extract with diverse phytoconstituents showing significant anticancer properties against the CACO2 cells with IC₅₀ value equal to 31.78 µg/mL. Nevertheless, all extracts showed potent anti-inflammatory activity at high concentrations (500 µg/mL). M. nodiflorum, M. forsskaolii, and M. cordifolium had IC₅₀ on HepG2 cells equal to 73.64, 88.18, and 87.82 µg/mL. Molecular findings showed the three extracts had distinct effects on apoptosis modulation in HepG2 cells. Conclusion The findings suggest that the studied extracts had potential therapeutic properties as anti-inflammatory and anticancer agents, supported by an in-silico interaction study.

Keywords: Colon cancer; GC-MS; HCC; Mesembryanthemum spp.; RBCs toxicity; anti-inflammatory.

Graphical abstract

Fig. 1

:GC-MS chromatograms of the aerial parts’ extracts of three Mesembryanthemum spp. A: TIC of M. nodiflorum; B: TIC of M. forsskaolii; C: TIC of M. cordifolium.

Fig. 2

Hemolysis percent of different concentrations of M. nodiflorum, M. forsskaolii, and M. cordifolium extracts compared to Diclofenac as a standard drug. (p-value= 0. 018).

Fig. 3

Percent inhibition of protein denaturation at different concentrations (62.5, 125, 250, 500, and 1000 µg/mL) for Diclofenac and extracts of M. nodiflorum, M. forsskaolii, and M. cordifolium. Statistical significance is indicated by asterisks: ***p < 0. 01, ***p < 0. 001, ****p < 0. 0001.

Fig. 4

Predicted Targets from SwissTargetPrediction. The pie chart illustrates the distribution of predicted targets classified by protein type of the four dominant compounds.

Fig. 5

Molecular docking of the four major compounds and VEGFR. The docking poses and the binding interactions of each compound with VEGFR are shown in the various panels. The yellow color represents the compound molecule in 2D and the white color represents the receptor; VEGFR.

Fig. 6

Results of the cell viability test based on varying concentrations of M. nodiflorum, M. forsskaolii, and M. cordifolium extracts compared to DOX as a reference drug. The line graph is a depiction of the extracts' effect on the Cell viability of both CACO2 and HepG2 cells. The M. forsskaolii extract showed a remarked viability decrease even with a lower concentration of the extract on CACO2 cells with IC₅₀ equal to 31.78 µg/mL. However, M. nodiflorum, and M. cordifolium extracts showed no observed IC₅₀. In HepG2 cells, the three extracts showed a closure calculated IC₅₀ values ranging from 73.64 to 88.18 µg/mL.

Fig. 7

Microscopic examination of HepG2 cells subjected to different extracts compared to untreated cells. The untreated cells had a confluent sheet however treated cells showed morphological changes including cell shrinking, decreased round size, and detached cells. (a): Untreated cells CACO2 cells, (b): Treated cells with M. forsskaolii at a concentration of 25 µg/mL, (c): Untreated HepG2 cells, (d): Treated cells with M. nodiflorum, (e): Treated cells with M. forsskaolii, and (f): Treated cells with M. cordifolium at a concentration equivalent to 100 µg/mL.

Fig. 8

Apoptosis-related gene expression in HepG2 cells treated with different extracts, M. nodiflorum, M. forsskaolii, and M. cordifolium. Gene expression levels of BCL-2, BID, BAX, PUMA & Cas-3 in untreated HepG2 cells and treated HepG2 cells. Statistically significant differences from untreated cells are indicated by *p < 0.05 and **p < 0.01.

Fig. 9

(a): Sandwich ELISA of α-fetoprotein in treated HepG2 cells compared to untreated. (b): LDH release detection in the treated HepG2 cells compared to untreated. p< 0.0001.