JAMES OSAKUE

Iron dysregulation remains a critical global health challenge, affecting billions through conditions ranging from iron deficiency anemia to hereditary hemochromatosis. This study investigated the effects of Piper guineense (Uziza) leaf extract on HFE gene, TFRC gene and survival rate of Drosophila melanogaster. Fresh Uziza leaves were collected, authenticated and extracted using 70% ethanol maceration. Five experimental groups (n=60 flies per replicate, three replicates each) were established: Control, 100 mg/mL, 200 mg/mL, 300 mg/mL, and 400 mg/mL Piper guineense extract treatments. Survival was monitored over 21 days, followed by molecular analysis using conventional PCR, agarose gel electrophoresis, and quantitative realtime PCR with Gpdh as the housekeeping gene. Results revealed a striking hormetic response pattern, with 100 mg/mL treatment significantly enhancing survival (66.67% ± 4.67%) compared to controls (48.33% ± 3.06%), representing 30.3% mortality reduction. Higher concentrations (300-400 mg/mL) demonstrated dose-dependent toxicity. HFE gene expression showed significant dose-dependent upregulation, peaking at 300 mg/mL (2.65-fold increase, p < 0.001), with progressive increases at 100 mg/mL (1.42-fold), 200 mg/mL (2.08-fold), and 400 mg/mL (1.98-fold) compared to controls. Statistical analysis (one-way ANOVA, F(4,10) = 6.82, p < 0.05) confirmed significant inter-group differences. The dissociation between optimal survival (100 mg/mL) and maximal gene expression (300 mg/mL) indicates that moderate transcriptional activation represents beneficial adaptation, while robust upregulation signals cellular distress. These findings demonstrate that Piper guineense bioactive compounds modulate iron metabolism gene expression through mechanisms likely involving iron chelation and oxidative stress signaling. The narrow therapeutic window emphasizes the critical importance of dose optimization for potential therapeutic applications in iron-related disorders.

Iron is one of the integral components of many biochemical properties which is maintained normal physiological activities for the healthy life. Insufficient iron causes different effects at the cellular level like limited oxygen supply, meager work performance and reduces immunity. The aim of this study is to determine the effect of the varied concentration of Gongronema latifolium leave extract on iron metabolism in Drosophila melanogaster. Samples were 1-2 days old virgin male and female Drosophila melanogaster fed with varied concentration of Utazi leaf extract supplemented corn meal diet. The control subjects include 1-2 days old virgin male and female Drosophila melanogaster fed with only corn meal diet and distilled water. The survival assay was carried out in three replicates of each concentration, for the determination of biochemical assays, a second group experiment was carried out. Ferritin ELISA kit was used to determined the Ferritin level, TIBC was determine using TIBC ELISA kit, serum iron was determined using serum iron ELISA kit. Results show that the average percentage survival was observed to be the highest in Group 5 having 80 percent of flies still alive at day 21. The Survival curve showed a negative correlation curve which shows that as the days increase, survival decreases. Group 1, the control group, exhibited a Ferritin level of 0.42±0.44, while Group 2, treated with aqueous Utazi extract, showed a slightly lower Ferritin level of 0.40±0.00. However, Group 3 (given Utazi extract), Group 4 (administered 5.0 mg Utazi extract), and Group 5 (given 2.5mg and 0.1mg of Utazi extract, respectively) demonstrated significant changes (p <0.001) in Ferritin levels, with Group 4 having the lowest level at 0.22±0.18. there was also highly significant difference (F=128.969, p=0.000*) in Ferritin levels among the group. Iron levels varied significantly among the groups, with Group 3 exhibiting the highest levels (16.18±18.46) and Group 2 the lowest (4.65±0.00). TIBC levels followed a similar trend, with Group 3 having the highest value (38.29±15.43) and Group 2 the lowest (6.33±0.00). The ANOVA analysis for TIBC showed significant differences (F=31.389, p=0.000*) among the groups. In conclusion, the study revealed a decreasing survival rate with increasing number of days that is a negative correlation between the concentration of Utazi leaf extract in the diet and on the survival of Drosophila melanogaster, suggesting potential toxicity at higher concentrations. Additionally, significant alterations in Ferritin, Iron, and Total Iron- Binding Capacity (TIBC) levels among experimental groups was observed, indicating that Utazi leaf extract can influence iron metabolism in fruit flies