Cytotoxic, Anti-Hemolytic, and Antioxidant Activities of Ruta chalepensis L. (Rutaceae) Extract, Fractions, and Isolated Compounds

Abstract

Ruta chalepensis is an herb used to treat various ailments, and its potential cytotoxic effects on different tumor cell lines have been extensively studied. The present study aimed to evaluate the cytotoxic activity of R. chalepensis methanol extract (RCME), sub-partitions obtained from solvents of increasing polarity, and major compounds, as well as their hemolytic, anti-hemolytic, and antioxidant potential. The in vitro cytotoxic activity against the human hepatocarcinoma (HEP-G2) and the murine lymphoma cell line (L5178Y-R) was evaluated using the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay, whereas selectivity indices (SIs) were determined by comparing cytotoxicity against normal African green monkey kidney cells (VERO) and human peripheral blood mononuclear cells (PBMC). Hemolytic and anti-hemolytic activities were evaluated on human erythrocytes. The most effective cytotoxic treatment was evaluated for nitric oxide release by J774A.1 macrophages. Antioxidant activity of R. chalepensis material was also determined. Results showed that RCME produced significant (p < 0.05) cytotoxicity in HEP-G2 (IC50 = 1.79 µg/mL) and L5178Y-R (IC50 = 1.60 µg/mL) cells and exhibited high SIs (291.50 and 114.80, respectively). In addition, the n-hexane fraction (RCHF) showed an IC50 of 18.31 µg/mL in HEP-G2 cells and an SI of 9.48 in VERO cells, whereas the chloroform fraction (RCCF) evidenced an IC50 of 1.60 µg/mL in L5178Y-R cells and an SI of 34.27 in PBMC cells. Chalepensin (CHL), rutamarin (RTM), and graveolin (GRV), which are major components of R. chalepensis, showed high activity against L5178Y-R cells, with IC50 of 9.15, 15.13 and SI of 45.08 µg/mL, respectively. In addition, CHL, RTM, and GRV showed SIs of 24.76, 9.98, and 3.52, respectively, when compared with PBMC cells. RCME at concentrations of 125 µg/mL and 250 µg/mL, significantly (p < 0.05) decreased nitrite production in J774A.1 cells, when exposed to lipopolysaccharide. This study demonstrated that RCME showed significant cytotoxic activity against HEP-G2 and L5178Y-R cells, without affecting normal VERO, PBMC, and J774A.1 cells.

Assessment of Anticancer Properties of Argemone mexicana L. and Berberine: A Comparative Study

“…The overproduction of NO causes tissue damage and is associated with chronic inflammation [90]. Some natural components present in herbal extracts have been shown to effectively inhibit LPS-induced NO in murine macrophages [38]. Therefore, as reported in Figure 5, BER exhibits immunomodulatory activity in response to NO production at concentrations of 0.98 to 500 µg/mL.…”

“…Prior to the application of the treatments, the cells were incubated at 37 • C in a humidified incubator (Sanyo MCO-19AIC CO 2 Incubator, Sanyo Electric Co., Ltd., Gunmaken, Japan) with 5% CO 2 for 24 h for adaptation [38]. The cell viability was determined by MTT assay; MTT color intensity was directly associated with the number of living cells [39] after 72 h of incubation.…”

“…The selectivity indexes (SI) were obtained after dividing the IC 50 of the normal cell on the IC 50 of the respective tumor cell. Any sample with an SI value greater than 3 was considered high [38]. Cell viability and SI were determined by the following formulas, respectively (2) The hemolytic activity was assessed using the hemolysis test [43].…”

“…The antioxidant activity was determined by the DPPH, ABTS radical scavenging [38], and FRAP (Ferric Reducing Antioxidant Power) [47] methods. In the DPPH and ABTS assays, Vitamin C served as the positive control.…”

“…The antioxidant activity was determined using the ABTS radical scavenging method [38], where the antioxidant activity (free radical scavenging capacity) was quantified as IC 50 in µg/mL. IC 50 represents the concentration of the test material required to cause a 50% decrease in the initial concentration of the ABTS radical.…”