In Vitro Study of the Phytochemical Composition and Antioxidant, Immunostimulant, and Hemolytic Activities of Nigella sativa (Ranunculaceae) and Lepidium sativum Seeds

Abstract

The Moroccan flora abounds and is an important reserve of medicinal plants. Nigella sativa and Lepidium sativum are plants that are widely used in traditional medicine for their multiple therapeutic properties. The current study aims to highlight the biological activities that can justify and valorize the use of these plants. Flavonoids, total phenols, condensed tannins, and sugars were determined. The biological activities tested were antioxidant by determining the IC50 (defined as the concentration of an antioxidant required to decrease the initial concentration by 50%; inversely related to the antioxidant capacity), hemagglutination, and hemolytic activities. Phytochemical quantification of the seed extracts indicated that the total phenol content was largely similar for both plants and in the order of 10 mg GAE (Gallic acid equivalent)/g. On the other hand, L. sativum seeds registered a higher content of flavonoids (3.09 ± 0.04 mg QE (quercetin equivalent)/g) as compared to Nigella saliva (0.258 ± 0.058). Concerning condensed tannins, N. saliva seeds present a higher amount with a value of 7.2 ± 0.025 mg/g as compared to L. sativum (1.4 ± 0.22 mg/g). Concerning the total sugar content, L. sativum shows a higher content (67.86 ± 0.87 mg/g) as compared to N. sativa (58.17 ± 0.42 mg/g); it is also richer in mucilage with a content of 240 mg as compared to 8.2 mg for N. saliva. Examination of the antioxidant activity using a DPPH (2.2-diphenyl 1-pycrilhydrazyl) test revealed that the EButOH (n-butanol extract) and EAE (ethyl acetate extract) extracts were the most active, with IC50 values of 48.7 and 50.65 μg/mL for the N. sativa extracts and 15.7 and 52.64 μg/mL for the L. sativum extracts, respectively. The results of the hemagglutination activity of the different extracts of the two plants prepared in the PBS (phosphate-buffered saline) medium showed significant agglutination for the L. sativum extract (1/50) compared to the N. sativa extract (1/20). An evaluation of the hemolytic effect of the crude extract of the studied seeds on erythrocytes isolated from rat blood incubated in PBS buffer compared to the total hemolysis induced by distilled water showed a hemolysis rate of 54% for Nigella sativa and 34% for L. sativum. In conclusion, the two plants studied in the current work exhibited high antioxidant potential, which could explain their beneficial properties.

Documentation of phytochemicals, bio-toxicity and haemato-physiological responses at sub-lethal concentrations of Parthenium hysterophorus hydroethanolic extract in Cyprinus carpio: a concept of waste valorization as a phyto-ichthyotoxin

“…Phyto-haemagglution activity and haemagglutination limit test in the present study also revealed the toxicity caused by the extract by causing clumping of the cells. Phytolectins in the extract are majorly responsible for the clumping of erythrocytes which ultimately hinders the normal functioning of the cells (Senthilganesh et al 2021;Ouattar et al 2022). A similar kind of agglutination was also reported by Abdali et al (2021) while working on Linum usitatissimum extract.…”

“…The test is based on the agglutination or precipitation of red blood cells caused by various toxic components present in the extract particularly lectins (Ouattar et al 2022). The assay was performed in a 96-well roundbottom plate as per the method described by Ouattar et al (2022).…”

“…The test is based on the agglutination or precipitation of red blood cells caused by various toxic components present in the extract particularly lectins (Ouattar et al 2022). The assay was performed in a 96-well roundbottom plate as per the method described by Ouattar et al (2022). Where, we mixed 25 µl of extract, 50 µl of NaCl solution (0.9%), and 25 µl blood suspension (3%) followed by incubation at 37°C for 30 min and observation of cells under the microscope for any agglutination of RBCs (red blood cells).…”

“…This study was conducted to nd out the minimum concentration which causes agglutination of erythrocytes by following the method described by Ouattar et al (2022). The assay was also performed in a 96-well roundbottom plate.…”

In-vivo Immunomodulatory Activities of Essential Oils of Artemisia abyssinica and Lepidium sativum in Mice

“… 20 , 21 Flavonoids, phenolics, alkaloids, saponins, glycosides, coumarins, fixed oils, proteins, vitamins, and minerals were found in L. sativum essential oil. 22 …”