Polydatin Increases Lifespan and Rescue Lead-induced Behavioural Deficits, Inflammatory and Oxidative Damage in Drosophila melanogaster Harwich Strain.

doi:10.21203/rs.3.rs-1970449/v1

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Polydatin Increases Lifespan and Rescue Lead-induced Behavioural Deficits, Inflammatory and Oxidative Damage in Drosophila melanogaster Harwich Strain.

https://doi.org/10.21203/rs.3.rs-1970449/v1

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Background

Lead (Pb) is toxic and cause many adverse clinical outcomes in children and adult, however, polydatin (PD) is a natural product from plants with reported antioxidative, neuroprotective and inflammatory properties. This study investigated the role of polydatin on lead-induced behavioural deficits, oxidative stress and inflammatory damages in D. melanogaster. D. melanogaster (Harwich strain, 1 to 3 days old) were orally administered lead acetate (PbAc) (0, 50, 100, 250 and 500 µM/5g diet) and PD (0, 5, 10, 20 and 40 µM/5g diet) for 14-days survival assays respectively. Thereafter, three concentrations of PD (10, 20 and 40 µM/5g diet) and one concentration of PbAc (250 µM/5g diet) were selected to evaluate the ameliorative potential of polydatin on PbAc-induced toxicity in D. melanogaster after 5-days oral co-treatment.

Results

Markers of behavioural (acetylcholinesterase, locomotor performance, fecundity and eclosure of the flies (emergence)), oxidative stress-antioxidant status (hydrogen peroxide, total thiol, catalase and glutathione-S-transferase, cell viability), inflammation (nitric oxide) were evaluated. Polydatin elevate the lifespan of D. melanogaster in a dose-dependent manner up to 40 µM/kg diet. Furthermore, polydatin alleviate PbAc-induced inhibition of catalase, glutathione-S-transferase and acetylcholinesterase activities in D. melanogaster. Moreover, polydatin significantly (p < 0.05) alleviate PbAc-induced cell death, behavioural deficits, accumulation of nitric oxide, hydrogen peroxide, total thiol levels and histopathological lesions in flies.

Conclusion

The lifespan prolonging effects of polydatin and its ameliorative role on PbAc-mediated toxicity in the flies may be due to its improvement in the behavioural deficits, antioxidant and anti-inflammatory properties.

Polydatin

Drosophila melanogaster

Eclosure

Lead toxicity

Locomotor performance

Over 240 million people are overexposed to lead and 99% of those with blood lead levels above 20 µg/dL are in the developing world [1]. Recently, the Institute for Health Metrics and Evaluation (IHME) recorded that 540,000 deaths and loss of 13.9 million healthy life, disability-adjusted life years (DALYs), due to lead poisoning [1, 2]. IHME analysis on the 2019 world burden of disease dataset found that approximately 900,000 individuals lost their lives due to poisoning by lead yearly [2]. Low and middle income countries, are faced with the challenge of this environmental toxicant e.g. in India alone, IHME recorded 4.6 million lead-attributable DALAYs and about 165,000 deaths were reported [3]. Chronic exposure to lead commonly causes haematological effects, such as anaemia, or neurological disturbances, including headache, irritability, lethargy, convulsions, muscle weakness, ataxia, tremors and paralysis [4, 5]. Acute exposures to lead cause gastrointestinal disturbances (anorexia, nausea and vomiting, abdominal pain), hepatic and renal damage, hypertension and neurological effects (malaise, drowsiness and encephalopathy) that may lead to convulsions and death [4, 6, 7]. The unregulated mining activities in Nigeria which resulted in serious toxicities and medical problems among adults and children alike are a source for concern. Unfortunately, the people involved have little or no education and therefore lack the ability to protect themselves from the ensuing toxicities and medical problem. Reports have so far indicated that children are more vulnerable to lead poisoning with no assess to medication due to high cost [8, 9, 2]. Over 400–500 children lost their lives as a result of acute lead poisoning accompanied with mining process Nigeria with pathway of exposure being food, water, soil and dust [10, 11, 12]. Lead exposure in water, soil and air are significant in several African countries [13]. In Nigeria, herbal remedies and eye cosmetics contaminated with lead are main channels of exposure among individuals [14, 15]. The number of persons with lead poisoning are growing globally and currently available chelating agents such as calcium disodium EDTA and dimercaptosuccinic acid (DMSA) are associated with adverse effects [16, 17]. Therefore, polydatin found in plants foods such as grapes, peanut, mulberries, Japanese knotweed (Polygonum cuspidatum), red wine, fruits and many food items, has more economical, therapeutic benefits and may provide lead compounds against lead poisoning. Polydatin could be a source of novel compounds with better efficacy and safety profile against lead poisoning. Polydatin is used traditionally against conditions associated with oxidative and tissue damage which might have occur as a result of lead poisoning [18] this requires scientific validation. Focusing on the continue investigation for more capable, economically accessible and affordable natural remedy for relieving lead poisoning abundant in plants, this study was therefore carried out to investigate the role of polydatin on lead-induced behavioural deficits, oxidative stress and inflammatory damages in D. melanogaster.

Effect of Lead Acetate and Polydatin on the Survival and Longevity Rate in D. melanogaster

Longtime administration of graded concentrations of polydatin (5, 10, 20 and 40 µM/5g diet) elevated longevity of flies by 6.06, 30.30, 42.42 and 51.51% respectively (Fig. 1). The amount of days that was equivalent to the period when half of flies (50%) dead were 22 days (6.06%), 24 days (30.30%), 24 day (30.30%) 29 days (42.42%) and 22 days (51.51%) from control and flies treated with 5, 10, 20 and 40 µM/5g diet of polydatin respectively. However, 10, 20 and 40 µM/5g diet of polydatin extend longevity of flies greater than other dose (i.e. 30.30, 42.42 and 51.51% respectively). Thereafter, these three concentrations were selected to determine the faith of Polydatin against PbAc-induced toxicity in the flies. Furthermore, on the 14 days survival rate of the administered PbAc, it was confirmed that there was significant (p < 0.05) reduction in survival rates of the flies when compared to control (Fig. 2A, B) and these reduction in survival rate were more prominent in the higher concentrations (250 and 500 µM, Fig. 2A, B). Therefore, 250 µM of PbAc concentration was selected for further study.

Longevity curve showing lifespan extending capacity of PD in D. melanogaster. PD (5, 10, 20, and 40 µM/5g diet) extended D. melanogaster lifespan by 6.06%, 30.30%, 42.42% and 51.51% respectively in comparison with the control. PD: Polydatin.

PbAc Survival curve (A and B) showing reduced survival rate of D. melanogaster exposed to PbAc. The decreases in survival were more pronounced in the higher concentrations (250–500 µM) than the lower concentrations (50 and 100 µM). PbAc: Lead Acetate.

3.2. Polydatin Alleviate PbAc-challenged Behavioural Deficit, Reduction of Fecundity, Eclosure of Offspring and Cell Viability

There was significant decline (p < 0.05) in the locomotor performance rate (Fig. 3A) in PbAc-induced flies along with significant (p < 0.05) inhibition of AChE activity (Fig. 3B) when compared with the control flies. Thereafter, co-treatment of PbAc and polydatin significantly (p < 0.05) restored inhibition of AChE activity and elevate the rate of locomotor performance of flies. Furthermore, the amount of eggs laid by the parent flies exposed to PbAc where significantly (p < 0.05) reduced when compared to control flies whereas parent flies co-treated with PbAc and polydatin at graded doses where significantly (p < 0.05) elevated when compared to PbAc-treated flies (Fig. 3C). In addition, the amount of young flies that eclosured after one day exposure of parent flies to PbAc and polydatin, as well as the cell viability of parent flies exposed to PbAc and polydatin for 5 days (Fig. 3D, E), was also evaluated. There was also significant (p < 0.05) reductions in flies' eclosure and cell viability in D. melanogaster exposed to PbAc whereas improvement in flies co-exposed to PbAc and polydatin was achieved (Fig. 3D, E).

Acetylcholinesterase activity (A), Negative Geotaxis (B), Fecundity (C), Eclosure (D) and Cell Viability (E) of D. melanogaster exposed to PbAc and PD for 5 days. Data are presented as Mean ± SEM of 50 flies/vial with 5 replicates per treatment group. ^a: Significant difference in relation to control group (ET 100µL) (p < 0.05); ^b: Significant difference in relation to control group (DW 100µL) and ^c: significant difference in relation to standard drug DMSA (0.05mg). ^d: significant difference in relation to PbAc (250µM). ^e: significant difference in relation to (PbAc + DMSA (0.05mg). PbAc: Lead Acetate, PD: Polydatin, DMSA: Dimercaptosuccinic Acid, ET: Ethanol, DW: Distilled Water.

3.3. Polydatin Alleviate PbAc-induced Accumulations of H₂O₂, T-SH and NO Levels and Inhibition of Catalase, GST and MAO Activities in Flies

Hydrogen peroxide and nitric oxide levels were assessed in D. melanogaster after 5 days of exposure to PbAc and it was observed that polydatin significantly (p < 0.05) restored PbAc-induced elevations of H₂O₂ (Fig. 4I) and NO levels (Fig. 4II). As a result of elevated levels of H₂O₂produced due to PbAc-induced in flies, selected markers of antioxidants such as total thiols (Fig. 4III), catalase (Fig. 4IV), GST (Fig. 4V) and MAO (Fig. 4VI) were also investigated. The levels of total thiols in PbAc and polydatin-exposed flies was significantly (p < 0.05) elevated when compared with the PbAc-induced flies. In addition, co-treatment of PbAc and polydatin restored PbAc-induced inhibition of catalase, GST and monoamine oxidase activities in D. melanogaster.

Hydrogen peroxide level (I), Nitric oxide level (II), Total thiols level (III), Catalase activity (IV), Glutathione-S-transferase activity (V) and Monoamine oxidase (VI) of D. melanogaster exposed to PbAc and polydatin for 5 days. Data are presented as Mean ± SEM of 50 flies/vial with 5 replicates per treatment group. ^a: Significant difference in relation to control group (ET 100µL) (p < 0.05); ^b: Significant difference in relation to control group (DW 100µL) and ^c: significant difference in relation to standard drug DMSA (0.05mg). ^d: significant difference in relation to PbAc (250µM). ^e: significant difference in relation to (PbAc + 0.05mg DMSA). PbAc: Lead Acetate, PD: Polydatin, DMSA: Dimercaptosuccinic Acid, ET: Ethanol, DW: Distilled Water.

3.4. Effect of Polydatin on Brain Histopathology in PbAc-induced Toxicity in D. melanogaster

Histopathology of the brains of flies exposed to PbAc and polydatin for 5 days is presented in Figure 5. Histology of the brain of control flies (A) and those treated with graded concentrations of polydatin (B, C and D) shows a normal feature of neurons (periphery) around the middle nerve fibres (white matter). Furthermore, the histopathology the brain of flies treated with PbAc (250 µM) (E) indicated eosinophilia and rarefaction of the white matter (dotted arrow), and segmental loss of neurons (arrows) in the brain hemispheres. Moreover, the histology of the brain of flies co-administered with PbAc (250 µM) + polydatin (10 µM /5g diet, (F) and PbAc (250 µM) + polydatin (20 µM /5g diet, (G) shows normal feature of the neurons (arrows). Finally, the histopathology of the brain of flies co-administered to PbAc (250 µM) + polydatin (40 µM/5g diet, (H) and PbAc (250 µM) + DMSA (0.05 mg/5g diet, (I) shows little eosinophilia of nerve fibres (dotted arrow) (Figure 5).

Brains of controls ET and DW(A, B), those exposed to DMSA (C) and PD (D, E and F) showed an even distribution of neurons (periphery) around the central nerve fibres (white matter); Histological lesions in the brain of PbAc-treated flies (G) showed eosinophilia and rarefaction of the white matter (dotted arrow), and segmental loss of neurons (red arrows) in the brain hemispheres; The histology of D. melanogaster co-treated with (H) PbAc (250 µM) + PD (10 µM/5g diet), (I) PbAc (250 µM) + PD (20 µM/5g diet) and (J) PbAc (250 µM) + PD (40 µM/5g diet) showed even distribution of the neurons (arrows); (K) PbAc (250 µM) + DMSA (0.05 mg/5g diet) showed slight eosinophilia of nerve fibres (red arrow). Magnification: X 400. PbAc: Lead Acetate PD: Polydatin, ET: Ethanol, DW: Distilled Water.

Lead has no important purpose in the animal and human body, but its accumulation in the body can result to unwanted effects, no matter it root of entrance [19]. Reactive oxygen species (ROS) can lead to cellular damage through manifestation of oxidative stress, which can result to many diseases like lead poisoning due to instability between pro-oxidants and antioxidants [20, 21]. Oxidative stress is a situation in which the manufacture of reactive species is mostly elevated and the available anti-oxidative protective mechanisms are insufficient, resulting in breaking down the carbohydrates, protein, DNA and lipids by many free radicals [22]. However, polydatin, a natural polyphenol with higher antioxidant activity found in abundant in plants have been tested against PbAc-induced toxicity in D. melanogaster for the first time. In this study, the ability of polydatin to increase life span and rescue PbAc-induced behavioral deficits, inflammation and oxidative damage in D. melanogaster was investigated.

Polydatin at the graded doses tested, significantly extended the lifespan of D. melanogaster in comparable to the control flies and the lifespan-extending capacity of the polydatin appeared be doses-dependent which implies that the higher the concentration the more the activity of polydatin in extending the lifespan of D. melanogaster. However, polydatin lifespan-extending capacity in flies found in this study may indicate that it possesses anti-aging activity. Aging is a process that result to physiological retardation and elevate changes to diseases such as Parkinson disease (PD) and ultimately result to death [23]. Although, the process of aging is cumbersome, researchers have revealed that age-related retardation in lifespan in flies is linked to many factors such as feeding on diets rich in antioxidant agents [24] which has been implicated in this study by polydatin. This finding is similar to the report of Baxter, [25] that resveratrol being natural polyphenolic compound possesses anti-aging property through activation of sirtuin.

Furthermore, markers of behavioural deficits were also evaluated in this study since PbAc-administered flies exhibited significant decline in negative geotaxis (locomotor performance) behaviour together with down-regulation of AChE activity. Acetylcholinesterase hydrolyses acetylcholine, a neurotransmitter that take part in an important activity of regulating locomotion and motor function [26]. As soon as PbAc cross blood brain barrier, it can interfere by inhibiting AChE activity thereby elevating acetylcholine level in the basal ganglia resulting to cellular miscommunication [27]. Therefore, the inhibition of AChE activity as confirmed in this study, might have distorted the normal neurotransmission of flies resulting to faulting locomotor performance of the flies. The evidence that polydatin alleviate PbAc-induced AChE inhibition and locomotor performance by this study, and the findings that neuroprotective agents can elevate locomotor performance of flies [28], enable the conclusion that polydatin have neuroprotective ability in D. melanogaster. Additionally, the observation that treatment of D. melanogaster with PbAc retard the egg laying ability of D. melanogaster (fecundity) and eclosure of offspring from pupae to adult (emergence) might be due to increased cell death. However, flies co-treated with polydatin and PbAc showed increased fecundity and emergence rate of flies comparable with control, thus suggesting its valuable role in D. melanogaster. This improvement by polydatin was more pronounce than the standard drug used in this study. The ability of polydatin to improve PbAc-induced retardation in fecundity and eclosion suggests that it may be effective in promoting reproduction.

Heme-containing enzyme known as Catalase [29] which is responsible for catalyzing the dismutaton of H₂O₂ to molecular oxygen and water, thereby reducing the chances of hydroxyl radical formation [30]. Moreover, PbAc caused H₂O₂ accumulation and inhibited catalase activity in D. melanogaster. Hydrogen peroxide combined with Fe²⁺ through Fenton reaction to bring about hydrogen oxide radical (HO.), a strong reactive oxidant [31]. Antioxidants provide defence to the cells by normalizing reactive oxygen species (ROS), but elevated intracellular make-up of ROS can lead to oxidative destruction to internal macromolecules like carbohydrates, proteins, lipids and nucleic acids, [32]. Thus, the fact that polydatin restored PbAc-induced inhibition of catalase activity and H₂O₂ level are indications that it possesses both antioxidative and free radical scavenging ability in D. melanogaster.

The phase II family of enzymes known as glutathione-S-transferase (GSTs) is responsible for catalyzing the conjugation of GSH with electrophiles. Furthermore, GSTs also participate in an important role in the survival of living organisms against oxidative destruction [33, 34]. Therefore, the confirmation that the activity of GST was inhibited in flies administered PbAc indicate that the enzyme was not activated in flies due toxicity caused by PbAc. However, polydatin at graded concentrations alleviate PbAc-induced inhibition of GST in flies which might have been due to its anti-oxidant potential. On the other hand, compounds with carbon-bound sulfhydryl group known as thiols (T-SH) [ 35] was also investigated in this study and the fact that PbAc significantly cause reduction in the level of T-SH may be partly due to augmentation of thiol-containing protein in the flies. Furthermore, polydatin significantly elevated the reduction of total thiol cause by PbAc in flies. This effect was seen to be more pronounced at the lower dose of polydatin which it effectiveness was more than that of the standard drug (DMSA), thus further confirming the antioxidative capacity of polydatin. Also, Monoamine oxidase was significantly inhibited by PbAc as observed in PbAc-administered flies but polydatin significantly elevated the activity of this enzyme even more pronounced than the flies co-treated with standard drug (DMSA). This further confirmed the antioxidant effect of polydatin in this study.

Moreover, polydatin prevented PbAc-induced elevation of NO level in flies. Nitric oxide take part in many physiological events. It is known as a pro-inflammatory mediator when it accumulates and reacts with superoxide anion to form more poisonous nitrite anion thereby resulting into different disease conditions [36]. The suppression of PbAc-mediated increase in NO concentration by polydatin indicate its ability to prevent inflammation from occuring in the flies. This suggest that polydatin possess anti-inflammatory activity.

In this present study, we also attempted to find out the role of cell death in PbAc-induced toxicity in flies. In this regard, it was confirmed that PbAc caused retardation in cell viability in the flies due to the fact that PbAc-administered flies showed reduction in cell viability. Furthermore, oxidative stress is on the other hand facilitated by MAO-B through mitochondrial release of cytochrome C and up-regulation of caspases 3 and 9, thereby causing neuronal destruction [37]. However, polydatin prevent cell damage by regulating viability of cells in flies.

Brain histopathological investigation was also carried out in this study, where PbAc-induced histopathological alterations was identified by eosinophilia and refraction of the white matter in the brain along with atrophy of neurons and nerve fibres. Thereafter, treatment with polydatin extensively diminished these lesions, thus corroborating to its protective role in PbAc-induced toxicity in D. melanogaster.

In conclusion, the lifespan extending effects of polydatin and its ameliorative role on PbAc-mediated toxicity in the flies may be due to its improvement in the behavioural deficits, antioxidant and anti-inflammatory properties.

Chemicals

All chemicals were of analytical grade. Polydatin (PD) (CAS: 27208-80-6) and lead acetate (98%, PbAc) were procured from Solarbio international trade limited, third floor, 85A, Lian dong U valley, tongzhou Dist. Beijing, China.

Ethical approval

All experimental protocols governing the care and use of experimental animals were considered. Ethical approval was obtained from ABU committee for animal uses and care (ABUCAUC/2021/147)

Fly Stock and Culture Medium

D. melanogaster (Harwich strain) was obtained, maintained and reared in Biochemistry Department, University of Ibadan and Department of Zoology, Ahmadu Bello University Zaria, Nigeria on cornmeal medium containing 1% w/v brewer's yeast, 2% w/v sucrose, 1% w/v powdered milk, 1% w/v agar, and 0.08% v/w nipagin at constant temperature and humidity (23 ± 1^oC; 60% relative humidity, respectively) under 12 h dark/light cycle conditions.

Determination of Survival and Longevity Rate Analysis on Lead Acetate and Polydatin Exposure

D. melanogaster (both sexes, 1-3 days of age) were randomly divided into different groups of 50 flies each and treated PbAc (0, 50, 100,250 and 500 µM/5g) and PD (0, 5, 10, 20 and 40 µM/5g diet) for 14 days survival and longevity assays in order to determine the concentrations and duration of exposure to PbAc and PD respectively. Daily death was recorded, analyzed and results was plotted as percentage of live flies. Based on these results, 10, 20 and 40 µM/5g doses of polydatin as well as 250 µM doses of PbAc were selected for exposure of flies for 5 days. Thereafter, markers of oxidative stress-antioxidant status (total thiol (T-SH), hydrogen peroxide (H₂O₂), catalase (CAT) and glutathione-S-transferase), neurotoxicity (locomotor performance and acetylcholinesterase (AChE)), inflammation (nitric oxide (NO), cell viability (3-[4, 5-dimethylthiazole-2yl]-2, 5-diphenyltetrazolium bromide (MTT, assay)), reproduction (fecundity assay and eclosure rate) were evaluated.

Determination of Locomotor Performance, Fecundity and Eclosure Rate of D. melanogaster

Negative geotaxis of polydatin and PbAc-treated flies was evaluated using the locomotor performance assay [38, 39]. Egg laying ability of the flies was carried out using method of Feany and Bender, [40]. Eclosure rate of D. melanogaster offspring after exposure to PbAc and polydatin was evaluated as reported by [41].

Sample Preparation for Biochemical Analysis

Fifty (50) D. melanogasters (of both sexes) were exposed to concentrations of PbAc (250 µM/5g diet), PD (10, 20 and 40 µM/5g diet), DMSA (0.05 mg/5g diet) and PbAc + PD of graded concentrations: (250 µM PbAc + 10 µM/5g diet PD); (250 µM PbAc + 20 µM/5g diet PD); (250 µM PbAc + 40 µM/5g diet PD), control (ethanol, 2% 100 µL) and (distilled water, 100 µL) in diet for 5 days. After the treatment period, D. melanogasters were anaesthetized using CO₂, weighed, homogenized in 0.1M phosphate buffer, pH 7.0 (ratio of 1 mg:10 mL), and centrifuged at 4000 g for 10 min at 4^oC in a Thermo Scientific Sorval Micro 17 R refrigerated centrifuge. Furthermore, the serums were separated from the pellets into labeled Eppendorf tubes, stored at -20^oC and used for the evaluations of GST, CAT and AChE activities as well as T-SH and H₂O₂ levels and cell viability. All the experiment were conducted in duplicates for each of the five replicates of PbAc and PD graded concentrations.

Determination of Effect of Polydatin on Biochemical Parameters in Lead-induced Toxicity in D. melanogaster

Acetylcholinesterase activity was investigated by the method of Ellman et al. [42]. Hydrogen peroxide level was established according to the method of Wolff [43]. Protein assayed was conducted by the method of Lowry et al. [44]. Total thiol content was evaluated by the method of Ellman [45]. Cell viability was carried out based on enzymatic retardation of MTT to MTT formazan at a final concentration of 5 mg/ml [46].Glutathione-S-transferase activity was assayed according to the procedure of Habig and Jakoby [47]. Catalase activity was determined using the method of Aebi [48]. Nitrite level in serum was carried out by the method of Griess reaction [49]. All the experiments were replicated at least two times in different institutions.

Determination of Effect of Polydatin on Histopathology of Brain in Lead-induced Toxicity in D. melanogaster

At the end of treatment, flies were anaesthesia with CO₂, tissues were incubated in 8% w/v paraformaldehyde for 8 hrs. After fixation, tissues were rinsed in PBS five times, each time for 10 min at room temperature. The tissue were dehydrated through an ethanol series to xylene. Xylene was replaced with paraffin in increasing volume at 62°C in an oven. Tissue was placed in hard paper mold containing molten paraffin, and stored at 0°C for at least 24 hrs blocks were mounted and sectioned at 7 µm thickness on a Yorco rotary microtome and slides containing sections were stored upright overnight at 37°C. Slides were processed in xylene for 10 min, followed by decreased ethanol series (100-30%). The sections were stained with Ehlich’s Haematoxylin for 2 min followed by 0.5% hydrochloric solution (1 dip), 0.5% sodium bicarbonate (5 min) and Scott’s solution (5 min). After that slides were stained with eosin for 10 min, rinsed with upgrade ethanol series (30-100%), followed by xylene for 10 min. Finally, the slides were mounted with DPX and observed under Leica microscope [50]. Detailed microscopic examinations were carried out by a consultant histopathologist. Photomicrographs of the organs were taken at magnification ( 100).

Statistical Analysis

Data were expressed as Mean ± Standard Error of the Mean (S.E.M.) and the differences between means were analyzed by One Way Analysis of Variance (ANOVA) for single point data followed by Dunnett post hoc test and Repeated Measure ANOVA for data collected over time followed by Bonferroni post hoc test. Graph pad prism version 8.0.2 was used for the analyses and values of p ≤ 0.05 was considered statistically significant. Results were presented in line graphs and charts; histopathological findings were presented as photomicrograph.

AChE: Acetylcholinesterase, Pb: Lead, PbAc: Lead acetate, PD: Polydatin, DMSA: Dimercaptosuccinic acid, DALYs: Disability-adjusted life years, IHME: Institute for Health Metrics and Evaluation, EDTA: Ethylene di amine tetra acetic acid, CAT: Catalase, GST: Glutathione-S-transferase, NO: Nitric oxide, MTT: (3-[4, 5-Dimethylthiazole-2yl]-2, 5-Diphenyltetrazolium Bromide), T-SH: Total thiol, H₂O₂: Hydrogen peroxide, ANOVA: Analysis of Variance, S.E.M: Standard Error of the Mean, MAO: Monoamine Oxidase, ROS: Reactive oxygen species.

Ethics approval and consent to participate

This study was approved by the ethical committee of Ahmadu Bello University, Zaria, Nigeria in accordance with the principles guiding the use and handling of experimental animals.

Consent for publication

Not applicable

Availability of data and material

All statistical results are stated in the article, and raw data and materials are available from the corresponding author on reasonable request.

Competing interests

The authors declared that they have no competing interest to his work.

Funding

No funding was received for this work.

Authors’ contributions

All authors contributed to the study conception and design. [Salisu Muhammad Highab] carried out the study; [Salisu Muhammad Highab], [Jamilu Ya’u] collected data, [Salisu Muhammad Highab], [Jamilu Ya’u] and [Muhammad Garba Magaji] carried out the data analysis and interpretation of results. The first draft of manuscript was written by [Salisu Muhammad Highab]. All authors commented on the first draft manuscript; [Jamilu Ya’u], [Muhammad Garba Magaji] and [Dalhatu Muktar Shehu] supervised the research and edited the manuscript for intellectual content. All the authors read and approved the final manuscript.

Acknowledgments

We thank Dr. Amos O. Abolaji of Biochemistry Department, University of Ibadan, Nigeria for giving us access to his laboratory space and equipment to carry out this research. We also thank Dr. Theophilus A. Jarikre of vetenary pathology Department, University of Ibadan, for his valuable contributions toward the histology of this work. We thank Drosophila Laboratory of Department of Zoology, Ahmadu Bello University Zaria, Nigeria for giving us the opportunity to use its facility.

Authors’ information

Mr. Salisu M. Highab, Department of Pharmacology, Faculty of Basic Medical Sciences, College of Medicine and Health Sciences, Federal University Dutse, Jigawa State, Nigeria. [email protected], +2347036357517.

Prof. Jamilu Ya’u, Department of Pharmacology and Therapeutics, Faculty of Pharmaceutical Sciences, Ahmadu Bello University, Zaria, Kaduna State, Nigeria. [email protected]m, +2348032415294

Prof. Muhammad G. Magaji, Department of Pharmacology and Therapeutics, Faculty of Pharmaceutical Sciences, Ahmadu Bello University, Zaria, Kaduna State, Nigeria. [email protected], +2348034685849

Prof. Dalhatu M. Shehu, Department of Zoology, Faculty of Life Sciences, Ahmadu Bello University, Zaria, Kaduna State, Nigeria. [email protected] +2348023666265

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Polydatin Increases Lifespan and Rescue Lead-induced Behavioural Deficits, Inflammatory and Oxidative Damage in Drosophila melanogaster Harwich Strain.

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