Immune-Boosting Potentiating Properties of Brassica nigra Hydroalcoholic Extract in Cyclophosphamide-Induced Immunosuppression in Rats

Abstract

The antioxidative and immune-boosting properties of the hydroalcoholic extract of Brassica nigra sprouts in cyclophosphamide-induced immunosuppression in rats were investigated in this study. B. nigra sprouts were prepared in the lab to monitor the bio-changes in bioactive compounds during the sprouting period up to 7 days at 17 ± 1 °C and 90% relative humidity. The total phenolic content (TPC), antioxidant activity (AOA), total flavonoids (TFs), total flavonols (TFLs), and total carotenoids (TCs) were evaluated. Consequently, the identification and quantification of phenolic acids, their derivatives, and flavonoids were carried out using HPLC. Subsequently, the selected BN sprout (6-day-old sprout) was biologically examined, and oxidative stress biomarkers, hematological parameters, immunoglobulins (Igs), and pro-inflammatory and anti-inflammatory cytokines were investigated. An increase in TPC, AOA, TFs, TFLs, and TCs was observed by increasing the sprouting time. The HPLC analysis indicated that the B. nigra seeds contained 10 phenolic acids and 4 flavonoids, predominantly syringic acid and quercetin, respectively. After 3 days, the number of phenolic acids increased to 16, predominantly syringic acid, and the number of flavonoids increased to 7, predominantly quercetin. On the 6th day, 13 phenolic acids were estimated, with the highest being benzoic acid, and 6 flavonoids were estimated, with the highest being quercetin. The greatest rise in phenols was seen on the sixth day of sprouting. These included caffeic acid, chlorogenic acid, cinnamic acid, ferulic acid, coumaric acid, benzoic acid, and rosmarinic acid. Flavonoids such as kaempferol and myricetin increased. The sprouts on day 6 were recorded as having the highest bioactive compounds and AOA content. The selected B. nigra sprouts were examined for antioxidative and immunomodulatory properties in a rat model. Dosing 250 and 500 mg kg−1, the rats exhibited significant improvements in terms of antioxidative stress and the number of white blood cells (WBCs), lymphocytes, and neutrophils in the blood, indicating stimulation of the immune response in a dose-dependent manner. In addition, the production of immune proteins, such as IgG, IgM, and IgA, was enhanced in the blood. Moreover, the 500 mg kg−1 concentration of BN extract stimulated cytokine production in a stronger manner than the 250 mg kg−1 concentration, indicating that the extract significantly increased immune activity. In conclusion, the results indicate that mustard seed extracts have immunosuppressive properties against cyclophosphamide-induced immunosuppression in rats.