Synergistic erythropoietic mechanisms of Chromolaena odorata and Tithonia diversifolia in the bone marrow of Wistar rats

Traditional treatment of paediatric anaemia in Nigeria entails the use of plants, of which Chromolaena odorata and Tithonia diversifolia are prominent. However, the underlying molecular pharmacology of these plant substances is yet to be established. This study aimed to elucidate the scientific mechanisms that underpin the traditional use of Chromolaena odorata and Tithonia diversifolia as remedy for paediatric anaemia. We investigated the impact of these plants on erythropoietin and erythropoietin receptor expression and monitored cyclin-dependent kinase inhibitors, and FasL/FAS signalling mechanism therefrom. Three samples were prepared from spouting leaves of C. odorata and leaves of flowering T. diversifolia as follows: C.O-group (100g C. odorata was blended in 100 ml distilled/deionized water (dd-water)), T.D-group (100g T. diversifolia was blended in 100 ml dd-water) and the T.D + C.O-group (50g each of C. odorata and T. diversifolia were blended in 100 ml dd-water). Eight-week old in-bred Wistar rats (female) were grouped into 4 ( n =3): basal control (2 ml of dd-water), C.O-group (30 mg/kg b.w. ), T.D-group (30 mg/kg b.w. ) and T.D + C.O-group (30 mg/kg b.w ). An oral dose of the extracts was administered 6-h for 72 h followed by RT-PCR analysis of erythropoietin (Epo), erythropoietin receptor (Epo-R), kip1.p27 (p27/cdkn1b), p21Waf1, Kip2-p57 and FAS/FASL. Western blot was used to investigate JAK2 phosphorylation in vitro in bone marrow primary culture. The administration of C. odorata and T. diversifolia potentiated the upregulation of erythropoietin. This was associated synergistically with an upshot of bone marrow erythropoietin receptor while p21Waf1, Kip2-p57 were downregulated in treatment groups. However, the combination of C. odorata and T. diversifolia did not evoke JAK2 phosphorylation in vitro in bone marrow. The combined administration of C. odorata and T. diversifolia results in the stimulation of erythropoietin expression and also the HIF-1α genes, though this does not occur via the erythropoietin receptor agonism.