Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology
Protective effect of Catharanthus roseus plant extracts against endosulfan and its isomers induced impacts on non-targeted insect model, Drosophila melanogaster and live brain cell imaging
Graphical abstract
Introduction
The USEPA classifies endosulfan as category I (highly acutely toxic) pesticide and WHO considers endosulfan as moderately hazardous (class II) to various organisms (Siddique et al., 2003). Due to the low water solubility, chemical stability, high lipid solubility, and slow rate of biotransformation, the endosulfan is considered as a potential environmental pollutant. Several experimental studies suggest that endosulfan has endocrine-disrupting potential, causes Parkinson's disease, behavioral effects, reproductive disorders, neuronal effects, premature aging as well as cancer (Ahmed and Ahmad, 1993; ATSDR, 1993; Dalsenter et al., 1999; Bisson and Hontela, 2002; Hiremath and Kaliwal, 2002; Wang, 2006).
Several researchers are focused on the inhibiting or ameliorating effect of various plant extracts against the damage caused by narrow spectrum pesticides. Plant compounds mediated toxicity reduction studies of xenobiotic like malathion, maneb, arsenic, diazinon, rotenone, etc. have been done extensively (Venkatesan et al., 2003; Anilakumar et al., 2009; Ahmed et al., 2008, Ahmed et al., 2010; Abdallah et al., 2012; Manfo et al., 2014; Banaee et al., 2015). However, the literature and understanding of the botanicals mediated minimization of endosulfan toxicity is presently limited. Although Catharanthus roseus is a well-studied medicinal plant with high antioxidant properties, only a few studies have addressed the toxicity reduction by using C. roseus extracts. All parts of the plant exhibit a high degree of therapeutical properties, mainly high antioxidative potency (Djeriidane et al., 2006; Mustafa and Verpoorte, 2007; Ferreres et al., 2008; Pereira et al., 2010; Ferrers et al., 2011). Besides antioxidant and anti-cancerous activity, the plant compounds exhibit anti-diabetic, anti-allergic, anti-inflammatory, anti-microbial, anti-helminthic, anti-ulcer, anti-diarrheal, wound healing, hypotensive, hypolipidemic, memory enhancement, cardioprotective and vasodilator activities (Pillay et al., 1959; Chattopadhyay et al., 1991; Sekar, 1996: Nayak et al., 2007; Patil and Ghosh, 2010; Rajput et al., 2011; Patel et al., 2011). The plant's secondary compounds are incorporated into a wide range of commercial and industrial applications, served as resources for flavors, aromas and fragrance, bio-based fuels and plastics, enzymes, preservatives, cosmetics, natural pigments, and bio-active compounds. The extracts of C. roseus act as a very strong inhibitor for mild steel corrosion (Shahba et al., 2016; Rana et al., 2017).
D. melanogaster, an insect model with well-documented genetics and developmental biology, is the closet invertebrate to humans and has been used for toxicological studies as well as for studying human diseases (Jeibmann and Paulus, 2009). Therefore, the present study aimed to investigate the protective effect of C. roseus extracts (aqueous and methanolic extracts) against endosulfan and its isomers induced impacts on non-targeted insect model D. melanogaster with live cell brain imaging. We address the following topics in the paper, such as, (1) which endosulfan (endosulfan total, alpha-endosulfan and beta-endosulfan) is highly toxic and how it alters the various developmental stages of D. melanogaster – hatchability, pupation, emergence pattern, body weight, reproduction, and survival rate; (2) bioactive properties of C. roseus – polyphenol content, Fe3+ reducing power, DPPH free radical scavenging efficacy, organic and inorganic content of extracts (aqueous and methanol); (3) toxicity of methanolic and aqueous plant extracts (eclosion inhibition concentration used as an index for estimating the toxicity of plant extracts); (4) protective effect of C. roseus extracts against 1/5th EC50 concentration of endosulfan and its isomers – evaluated by monitoring the life stage performance recovery and protein profile variations (5) protective effects of C. roseus extracts validated by using live brain cell imaging of D. melanogaster using mitochondrial (TMRM) and nuclear stain (Hoechst).
Section snippets
Endosulfan total and its isomers
Endosulfan (C9H6C16O3S) is a polycyclic chlorinated hydrocarbon insecticide, it is a mixture of two stereoisomers, alpha-endosulfan (Endosulfan I; exo-configuration), and beta-endosulfan (Endosulfan II; endo-configuration). The technical grade of Endosulfan total and its isomers (α and β endosulfan) were procured from Sigma-Aldrich C. USA (Endosulfan 32015, Lot #SZBC166XV, Alpha-endosulfan 45468, Lot #SZBD205XV and Beta-endosulfan 33385, Lot #SZBD247XV).
Determination of effective concentration (EC50) of endosulfan and its isomers
The culture media prepared according to
EC50 concentration of endosulfan and its isomers
From the definitive test we arrived at the five different concentration of endosulfan (total) and alpha-endosulfan (2.28 ppm, 3.08 ppm, 4 ppm, 5.28 ppm, and 7 ppm) and beta endosulfan (2.5 ppm, 3.52 ppm, 5 ppm, 7 ppm, and 10 ppm). Toxicity response, EC50 in D. melanogaster against alpha-endosulfan, beta-endosulfan and endosulfan (total) was performed; 95% of confidence limits were obtained by strait line graphic interpolation methods of Litchfield and Wilcoxon (1949). The EC50 value of
Discussion
Several studies have emphasized that endosulfan shows acute and chronic toxicity in insects and mammals including human beings. The short-term toxicity of endosulfan can cause nervous system dysfunction, hyperactivity, decreased respiration, anemia, etc. it also causes vomiting, seizures, convulsions, and loss of consciousness, leading to death (Smith, 1991). The long-term exposure of endosulfan could be harmful to the central nervous system, immune system, and reproductive system. Endosulfan
Conclusion
The present study has undertaken to investigate the toxic effect of endosulfan and ameliorating effects of aqueous and methanol extracts of C. roseus. Endosulfan induced toxicity impact on D. melanogaster changed the C. roseus extract by increasing reduced body weight, increasing reduced life span, and overcome the developmental delay and also the reappearance of normal protein profile. Several studies have proved that the secondary metabolites of plants can cause a reduction of toxicity
Declaration of competing interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgments
We thank the Department of Zoology, University of Calicut for providing the infrastructural facility. And also, Dr. T. R. Santhosh Kumar, Scientist G, Rajiv Gandhi Centre for Biotechnology (RGCB) for providing valuable suggestions and laboratory facilities for doing live cell imaging. The first author is thankful to the Drosophila stock center, Department of Zoology, Mysore University for providing the model organism (D. melanogaster). This study financially supported by UGC – SAP, Government
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