Nano-Pelargonidin Protects Hyperglycemic-Induced L6 Cells against Mitochondrial Dysfunction

Abstract

Nano-encapsulation of several natural products has become an important tool in enhancing the bioavailability of some modern drugs against many diseases. Pelargonidin is an anthocyanidin found in many fruits and vegetables. Pelargonidin is loaded with poly-lactide-co-glycolic-acid, a non-toxic biodegradable polymer, to produce nano-pelargonidin. Size, morphology, zeta potential, and planar uniformity of formulated nano-pelargonidin were determined by atomic force microscopy and dynamic light scattering. The time required for cellular entry, folds of nano-pelargonidin, and drug encapsulation efficiency of poly-lactide-co-glycolic-acid were also ascertained. Relative functional efficacy of nano-pelargonidin and pelargonidin was evaluated by examining markers such as pyruvate kinase, glucokinase, calcium ion level, ATP/ADP ratio, mitochondrial membrane potential, cytosolic release of mitochondrial cytochrome-c, and structural analysis of mitochondrial DNA in controlled and experimental sets of alloxan-induced hyperglycemic L6 cells. Expressions of mitochondrial apoptotic proteins, such as bcl2 and caspase3, and glucose signalling cascades, such as GLUT4, IRS1, IRS2, and PI3, were analyzed. Nano-pelargonidin at a nearly 10-fold reduced dose significantly enhanced protection, presumably due to its smaller size, ability of faster entry, and drug delivery at target-specific sites. Thus, nano-pelargonidin can be used in formulating protective drugs for therapeutic management of mitochondrial dysfunction often encountered in diabetic conditions.

Key words

nano-technology - pelargonidin - atomic force microscopy - mitochondrial dysfunction - L6 cells

• References

• 1 Samadder A, Abraham SK, Khuda-Bukhsh AR. Nanopharmaceutical approach using pelargonidin towards enhancement of efficacy for prevention of alloxan-induced DNA damage in L6 cells via activation of PARP and p53. Environ Toxicol Pharmacol 2016; 43: 27-37
• 2 Samadder A, Das S, Das J, Khuda-Bukhsh AR. Relative efficacies of insulin and poly (lactic-co-glycolic) acid encapsulated nano-insulin in modulating certain significant biomarkers in arsenic intoxicated L6 cells. Colloids Surf B Biointerfaces 2013; 109: 10-19
• 3 Khandelwal N, Abraham SK. Protective effects of common anthocyanidins against genotoxic damage induced by chemotherapeutic drugs in mice. Planta Med 2014; 80: 1278-1283
• 4 Samadder A, Das J, Das S, De A, Saha SK, Bhattacharyya SS, Khuda-Bukhsh AR. Poly(lactic-co-glycolic) acid loaded nano-insulin has greater potentials of combating arsenic induced hyperglycemia in mice: some novel findings. Toxicol Appl Pharmacol 2013; 267: 57-73
• 5 Maechler P, Woheim CB. Mitochondrial function in normal and diabetic β-cells. Nature 2001; 414: 807-812
• 6 Patel N, Khayat ZA, Ruderman NB, Klip A. Dissociation of 5′ AMP-activated protein kinase activation and glucose uptake stimulation by mitochondrial uncoupling and hyperosmolar stress: differential sensitivities to intracellular Ca2+ and protein kinase C inhibition. Biochem Biophys Res Commun 2001; 285: 1066-1070
• 7 Samadder A, Chakraborty D, De A, Bhattacharyya SS, Bhadra K, Khuda-Bukhsh AR. Possible signaling cascades involved in attenuation of alloxan-induced oxidative stress and hyperglycemia in mice by ethanolic extract of Syzygium jambolanum: drug-DNA interaction with calf thymus DNA as target. Eur J Pharm Sci 2011; 44: 207-217
• 8 Fessi H, Puisieux F, Devissaquet JP, Ammoury N, Benita S. Nanocapsule formation by interfacial polymer deposition following solvent displacement. Int J Pharm 1989; 55: 1-4
• 9 Samadder A, Das S, Das J, Paul A, Khuda-Bukhsh AR. Ameliorative effects of Syzygium jambolanum extract and its poly (lactic-co-glycolic) acid nano-encapsulated form on arsenic-induced hyperglycemic stress: a multi-parametric evaluation. J Acupunct Meridian Stud 2012; 5: 310-318
• 10 Kaplan LA. Carbohydrates and Metabolite. In: Kaplan LA, Pesce AJ. eds. Clinical Chemistry: Theory, Analysis and Co-Relation. Toronto: C.V. Mosby; 1984: 1032-1040
• 11 Burnette WN. “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate – polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein. Anal Biochem 1981; 112: 195-203
• 12 Das J, Das S, Paul A, Samadder A, Khuda-Bukhsh AR. Strong anticancer potential of nanotriterpenoid from Phytolacca decandra against A549 adenocarcinoma via a Ca²⁺-dependent mitochondrial apoptotic pathway. J Acupunct Meridian Stud 2014; 7: 140-150
• 13 Schafer ZT, Grassian AR, Song L, Jiang Z, Hines ZG, Irie HY, Gao S, Puigserver P, Brugge JS. Antioxidant and oncogene rescue of metabolic defects caused by loss of matrix attachment. Nature 2009; 461: 109-113
• 14 Zimmerman EG, Akins DR, Planz JV, Schurr MJ. A rapid procedure for isolating mitochondrial DNA. Gene Anal Tech 1988; 5: 102-104