PHYTOCHEMICAL PROFILING, CYTOTOXIC, ANTIOXIDANT AND IMMUNOSTIMULATORY EFFECTS OF CYPERUS ROTUNDUS: EVIDENCE-BASED VALIDATION OF A TRADITIONAL MEDICINAL PLANT

Abstract

Background: Cyperus rotundus, a traditional medicinal plant, has been extensively studied for its phytochemical composition and therapeutic properties, including its anticancer and anti-inflammatory activities. Cyperus rotundus is used in traditional medicinal systems for the treatment of diarrhea, dysentery, dysmenorrhea, and menstrual irregularities. This study aimed to investigate the phytochemical profile and cytotoxic, antioxidant, and immunostimulatory effects of Cyperus rotundus extract. Methods: Ethanol extract (CRE) and subsequent dichloromethane (CRD) and aqueous (CRA) fractions were prepared from Cyperus rotundus using the Soxhlet method.  Various methods have been used to screen phytochemicals. The antioxidant capacity of the extracts was assessed using the DPPH assay. The cytotoxic potential of the extracts was evaluated in Hep2 cells using an MTT assay. The immunomodulatory effects of Cyperus rotundus were examined using flow cytometry-based surface immunophenotyping, focusing on the activation markers CD80 and CD86 as well as Intracellular Cytokine Staining for Interleukin-6 and Tumor Necrosis Factor-α. Results: Phytochemical screening revealed the presence of alkaloids, phenolics, saponins, glycosides, steroids, phytosterols, flavonoids, and carbohydrates in the extracts. The aqueous extract showed minimal cytotoxicity on Hep2 cells with an IC50 of 189.8 μg/mL, while the ethanolic and dichloromethane extracts had IC50 values of 372.5 μg/mL and 263.7 μg/mL, respectively. The aqueous extract exhibited the highest DPPH radical scavenging activity (IC50 = 310.8 μg/mL), followed by the ethanolic extract (IC50 = 591.2 μg/mL). Gas chromatography-mass spectrometry (GC-MS) analysis of the aqueous extract identified forty-three compounds. Flow cytometry analysis demonstrated that the aqueous extract significantly enhanced the activation of monocytes, monocyte-derived dendritic cells (MoDCs), and plasmacytoid dendritic cells (pDCs) by upregulating the expression of the costimulatory molecules CD80 and CD86. Intracellular cytokine staining revealed increased production of IL-6 and TNF-α in monocytes upon treatment with the aqueous extract. Conclusion: These findings validate the ethnomedicinal use of Cyperus rotundus as an immunomodulator and highlight its potential as a safe phytomedicine for managing oxidative stress and immune dysfunction