Evaluation of in vitro antioxidant, anti-bacterial, cytotoxic and in vivo analgesic and neuro-pharmacological investigation of alysicarpus vaginalis available in Bangladesh

Abstract

Purpose: The objectives of this research were to investigate the effects of Alysicarpus vaginalis L. (Alyce clover) leaf extracts prepared with ethyl acetate and methanol on in vitro activities like antioxidant, antimicrobial and cytotoxic, as well as on some in vivo activities like analgesic and neuropharmacological properties in an animal model. Study Design: Phytochemical screening was used as the primary method of evaluation for both the ethyl acetate (EAEAV) and methanolic extract (MEAV) of Alysicarpus vaginalis (L.). Because of the pharmacological interest in the plant's constituent components, it was decided to investigate its potential on in vitro activities, such as antioxidant, cytotoxic and antibacterial. Whether or if the observed effects on its in vivo analgesic and neuropharmacological properties in animal models are statistically significant. Place and Duration of Study: This investigation was performed in Laboratory of Phytochemistry and Pharmacology, Department of Pharmacy and Laboratory of Microbiology, Stamford University Bangladesh, Dhaka between June 2022 to October 2022. Methodology: Both EAEAV and MEAV was subjected for phytochemical screening using various test reagent and was also researched for its potential antioxidant, antibacterial, cytotoxic, analgesic, and neuropharmacological effects. The DPPH Free Radical Scavenging Assay, the Disk Diffusion Method and the Brine Shrimp Lethality Bioassay were all used to determine cytotoxic, antimicrobial and antioxidant activity, using different concentrations respectively. Swiss albino mice were subjected to acetic acid-induced writhing and hot plate tests to determine analgesic efficacy. cGMP pathway induced mechanism test was used to confirm the actual analgesic activity. Open field and hole cross tests were used to evaluate locomotor activity as part of the neuropharmacological experiment. Results: Different phytochemical constituent were detected in the experimental result of EAEAV and MEAV's phytochemical screening. MEAV showed highest antioxidant activity with an IC50 value of 34.70 μg/mL where EAEAV showed IC50 value of 12.49 μg/mL and MEAV also showed higher antibacterial activity compared to EAEAV. EAEAV and MEAV showed most notable lethality with LC50 value of 3.89 μg/mL and 8.71 μg/mL compared to the conventional vincristine sulphate's LC50 value in the brine shrimp lethality bioassay. As opposed to this, EAEAV reduced peripheral nociception in the acetic acid-induced writhing nociceptive paradigm, with percent inhibitions of 42.48% and 47.00% where MEAV reduced 40.63% and 51.77% at the aforementioned dosages as opposed to conventional Diclofenac sodium. cGMP pathway mechanism confirmed the % inhibition of pain using both EAEAV which boosted the pain reduction into 59.62% and 71.12%. Additionally, a considerable analgesic effect was seen in the hot plate test. The extract, when compared to the usual Diazepam, was also shown to reduce motor coordination in the Open Field and Hole Cross Tests. Conclusion: These results confirmed the conventional medicine claims of the Alysicarpus vaginalis (L.) plant's therapeutic virtues and suggest potential mechanisms of action.