Daniel Lotanna Ajaghaku1,2, Peter Achunike Akah1,3, Emmanuel Emeka Ilodigwe1, Sunday Odunke Nduka4, Uduma Eke Osonwa5, Festus Basden Chinedu Okoye6. 1. a Department of Pharmacology/Toxicology, Faculty of Pharmaceutical Sciences , Nnamdi Azikiwe University , Awka , Nigeria. 2. b Department of Pharmacology , Enugu State University of Science and Technology , Ebeano , Enugu State , Nigeria. 3. c Department of Pharmacology/Toxicology, Faculty of Pharmaceutical Sciences , University of Nigeria , Nsukka , Nigeria. 4. d Department of Clinical Pharmacy and Pharmacy Management, Faculty of Pharmaceutical Sciences , Nnamdi Azikiwe University , Awka , Nigeria. 5. e Department of Pharmaceutics and Pharmaceutical Technology, Faculty of Pharmaceutical Sciences , Nnamdi Azikiwe University , Awka , Nigeria. 6. f Department of Pharmaceutical and Medicinal Chemistry , Nnamdi Azikiwe University , Awka , Nigeria.
Abstract
BACKGROUND: Millettia aboensis (Hook. F.) Baker (Fabaceae) is popular in ethnomedicine for its acclaimed efficacy in a number of disease conditions. This study evaluated the immunomodulatory effect of the leaf extract as a possible mechanism of its ethnomedicinal uses. METHODS: Humoral and cellular immune responses of Balb/c mice to tetanus toxoid and cyclophosphamide, respectively, were used to monitor immunomodulatory activities of the ethanol leaf extract and fractions of M. aboensis at 200, 300 and 400 mg/kg. Active (butanol) fraction of the extract was subjected to chromatographic purifications to isolate the active compound and the structure elucidated by a combination of 1D and 2D NMR and mass spectrometry. Stimulation of specific T-lymphocytes using intracellular cytokine staining technique was used to evaluate immune-enhancing activity of the isolated compound. RESULTS: The extract and fractions evoked increase in both humoral and cellular immunity. At 400 mg/kg of butanol fraction, the normalized mean secondary production of IgG1 and IgG2a antibodies were 9.0 and 7.7, respectively. Serum cytokine production by butanol fraction following secondary challenge with tetanus toxoid showed that IL-12, IL-17A and IFN-γ were expressed by 48.14, 41.37 and 38.22%, respectively. Structural elucidation of the active compound revealed presence of isomeric mixtures of quercetin-3-O-rutinoside and quercetin-3-O-robinobioside (Compound 1a/b). Compound 1a/b exhibited in vitro upregulation of specific CD4+ T-lymphocytes that were largely IFNγ releasing with up to 43.7% stimulation at 6.25 μg/mL compared to the baseline effect in DMSO vehicle control group. CONCLUSION: M. aboensis expressed strong immune-enhancing properties, which may explain its ethnopharmacological use in disease management.
BACKGROUND: Millettia aboensis (Hook. F.) Baker (Fabaceae) is popular in ethnomedicine for its acclaimed efficacy in a number of disease conditions. This study evaluated the immunomodulatory effect of the leaf extract as a possible mechanism of its ethnomedicinal uses. METHODS: Humoral and cellular immune responses of Balb/c mice to tetanus toxoid and cyclophosphamide, respectively, were used to monitor immunomodulatory activities of the ethanol leaf extract and fractions of M. aboensis at 200, 300 and 400 mg/kg. Active (butanol) fraction of the extract was subjected to chromatographic purifications to isolate the active compound and the structure elucidated by a combination of 1D and 2D NMR and mass spectrometry. Stimulation of specific T-lymphocytes using intracellular cytokine staining technique was used to evaluate immune-enhancing activity of the isolated compound. RESULTS: The extract and fractions evoked increase in both humoral and cellular immunity. At 400 mg/kg of butanol fraction, the normalized mean secondary production of IgG1 and IgG2a antibodies were 9.0 and 7.7, respectively. Serum cytokine production by butanol fraction following secondary challenge with tetanus toxoid showed that IL-12, IL-17A and IFN-γ were expressed by 48.14, 41.37 and 38.22%, respectively. Structural elucidation of the active compound revealed presence of isomeric mixtures of quercetin-3-O-rutinoside and quercetin-3-O-robinobioside (Compound 1a/b). Compound 1a/b exhibited in vitro upregulation of specific CD4+ T-lymphocytes that were largely IFNγ releasing with up to 43.7% stimulation at 6.25 μg/mL compared to the baseline effect in DMSO vehicle control group. CONCLUSION: M. aboensis expressed strong immune-enhancing properties, which may explain its ethnopharmacological use in disease management.
Authors: Manikya Pramudya; Firli Rahmah Primula Dewi; Richard W Wong; Devinta Wahyu Anggraini; Dwi Winarni; Sri Puji Astuti Wahyuningsih Journal: Vet World Date: 2022-05-12