Evaluation of pyruvate kinase and glucose 6 phosphate dehydrogenase mRNA in Drosophila melanogaster administered aqueous Datura stramonium leaf extract

Abstract

Background: This study investigated the dose-dependent effects of aqueous Datura stramonium leaf extract on the expression of pyruvate kinase (pyk) a critical glycolytic enzyme and glucose-6-phosphate dehydrogenase (G6PD).

Methods: The flies (50 flies/vial each performed in triplicates) were divided into four groups according to the different concentrations of D. stramonium extract. Adult flies were allocated to a control group (no extract) and three treatment groups (200, 400, and 800 mg/ 10g diet). The extract was administered to the flies ad libitum. Flies were collected into TRIzol reagent for preservation of mRNA and subsequently homogenized for analysis. Semi quantitative PCR was used to analyze the expression pattern of the genes and ImageJ was used to quantify the different bands on the PCR and GAPDH was used as the housekeeping gene. ANOVA and turkey post Hoc test was then used to determine the statistical significance between groups (p<0.05).

Results: Semi-quantitative analysis revealed that the extract significantly repressed the expression of both pyk and G6PD genes in all treated groups compared to controls. The repression of pyk was greater in the 800 mg/10g diet group (34.14±1.65) compared to the control (48.35±1.13, p<0.05). Similarly, G6PD expression was significantly reduced, particularly in the 200 mg/10g diet group (77.84±1.54) compared to the control (97.15±1.23, p<0.01)

Conclusion: These findings demonstrate that D. stramonium extract elicited significant, dose-dependent repression of both genes across all treatments relative to controls. Thereby possibly disrupting core metabolic pathways which play roles in glycolysis and antioxidant defense.