Elsevier

Chemico-Biological Interactions

Volume 256, 25 August 2016, Pages 25-36
Chemico-Biological Interactions

A novel therapeutic application of solid lipid nanoparticles encapsulated thymoquinone (TQ-SLNs) on 3-nitroproponic acid induced Huntington’s disease-like symptoms in wistar rats

https://doi.org/10.1016/j.cbi.2016.05.020Get rights and content

Highlights

  • Targeted drug delivery system against 3-NP induced HD has been developed.

  • 3-NP causes behavioral deficits via striatal lesions and oxidative stress.

  • TQ-SLNs treatment restored the behavioral despairs and oxidative injury than TQ-S.

  • TQ-SLNs also protected the striatal structural microelements from 3-NP toxicity.

  • Study signifies TQ-SLNs a potent neuroprotective formulation upon 3-NP induction.

Abstract

Huntington’s disease (HD), a devastating neurodegenerative disease causing a remarkable pathogenesis involves mitochondrial dysfunction and bioenergetics failure. 3-Nitropropionic acid (3-NP) is a unique toxin model of HD that are mainly confined to mitochondrial complex-II inhibition and free radical generation. Recently, several nanoparticle formulations were developed to treat against various neurodegenerative diseases including HD. One among them is solid lipid nanoparticles (SLNs), a colloidal carrier designed to enhance the brain drug delivery and to prolong the bio-availability of drugs in the system. Hence, the present study was framed to evaluate solid lipid nanoparticles encapsulated thymoquinone (TQ-SLNs) in comparison with thymoquinone suspension (TQ-S) against 3-NP induced behavioral despair, oxidative injury and striatal pathology. This study reports that theTQ-SLNs (10 and 20 mg/kg) and TQ-S (80 mg/kg) treated animals showed a significant (P < 0.01) improvement in the muscle strength, rigidity, movement and memory performances on 7th and 14th day behavioral analysis than TQ-S (40 mg/kg) treated group. Similarly, TQ-SLNs highly attenuated the levels of oxidative stress markers such as LPO, NO and protein carbonylsin 3-NP induced animals. Further, TQ-SLNs significantly restored the antioxidant defense system, controls the mitochondrial SDH inhibition and alleviates anti-cholinergic effect upon 3-NP induction. In addition, TQ-SLNs efficiently protected the striatal structural microelements against 3-NP toxicity, which was confirmed by light microscopic studies. Thus, the present investigation, collectively suggests that the low dose of TQ-SLNs supplementation is highly sufficient to attain the effect of TQ-S (80 mg/kg) to attenuate behavioral, biochemical and histological modifications in 3-NP exposed HD model.

Introduction

Huntington’s disease (HD) is an incurable, progressive neurodegenerative disease characterized by chorea, seizures, involuntary movements, dystonia, cognitive decline and emotional disturbances [1]. The hallmark of HD pathogenesis includes excitotoxic events, oxidative stress, impaired energy metabolism, mitochondrial dysfunction, altered synaptic transmission and transcriptional dysregulation [2]. The mechanistic background of HD remains unclear, nevertheless mutant htt protein (huntingtin protein with poly CAG repeats) largely aggregate in the caudate nucleus and putamen of basal ganglia and causes degeneration [3], [4].

Experimentally HD can be mimicked by an environmental mycotoxin 3-nitropropionic acid (3-NP) that efficiently replicates both the neurobehavioral alterations and striatal lesions [5]. 3-NP irreversibly inhibits succinate dehydrogenase (SDH) enzyme located in the inner domain of mitochondrial membrane, hence alters the mitochondrial electron transport chain and Krebs cycle, which leads to electron leakage from the mitochondria, bioenergetics failure and the generation of reactive oxygen species (ROS) and reactive nitrogen species (RNS) [6]. In addition, systemic application of 3-NP evoked a selective striatal loss closely resembling lesions that observed in the course of Huntington’s disease [5], [7].

Nigella sativa L., commonly known as black seed or black cumin, has been used in folk medicine as a natural remedy for the conditions such as respiratory illness, stomach and intestinal problems, kidney and liver functions, circulatory and immune system support, and for general well-being [8]. Most of the biological activity of the seeds has been shown to be due to thymoquinone (TQ) (2-isopropyl-5-methyl-1,2-benzoquinone), the major component of volatile oil [9]. Interestingly, this pharmacologically active lipophilic quinone is greatly responsible for many of the seed antioxidant and anti-inflammatory effects [10], hence it is frequently used in herbal medicine for treating various ailments. Furthermore, it has numerous beneficial properties including anti-inflammatory actions [11], neuroprotection [12] and suppression of oxidative stress-induced neuropathy [13]. TQ also improves neuronal morphology and reduces its degeneration during chronic toluene exposure [14], further it shows significant anti-anxiety like activity by modulating the levels of nitric oxide (NO) and γ-amino butyric acid (GABA) [15], and reduces nuclear factor kappa B (NF-KB) activation in brain and spinal cord [16].

Despite of various therapeutic efficacy of TQ, this compound suffers from poor solubility and high hydrophobicity leading to poor formulations for pharmaceutical applications [17]. This problem was solved by nanoparticulate encapsulation system. Solid lipid nanoparticles are emphasized to be a suitable colloidal carrier for the delivery of hydrophobic drugs to the central nervous systems (CNS) [18]. Considerably, SLNs were found to promote the stability, absorption and pharmacokinetic properties of the drug. In addition, it also improves drug payload, sustained drug release ability and drug targeting potential [19], [20]. Earlier we have formulated and characterized thymoquinone encapsulated solid lipid nanoparticles (TQ-SLNs) [21], further the developed formulations were found to be spherical shaped structure with the particle size of 172.10 ± 7.41 nm and maximum drug entrapment of 84.49 ± 3.36%. Also, drug release potential of 86.15 ± 2.76% for 72 h has been derived. Consequently, Singh et al. [22]reported that there was 5-fold increase in bio-availability of TQ when incorporated into SLNs. Hence, further studies have been attempted for the first time to evaluate the pharmacodynamic efficacy of developed solid lipid nanoparticles encapsulated thymoquinone (TQ-SLNs) in comparison with thymoquinone suspension (TQ-S) in alleviating 3-NP induced body weight change, behavioral impairments, oxidative cell damage and striatal cell pathology.

Section snippets

Chemicals and reagents

3-Nitropropionic acid and Thymoquinone were purchased from Sigma Chemical Co. (St. Louis, USA). Stearic acid, Soy Lecithin and Sodium taurocholate were purchased from Himedia Laboratories Pvt. Ltd. (Mumbai, India). Glutathione reduced form (GSH), glutathione oxidized form (GSSG), 1-chloro-2, 4-dinitrobenzene (CDNB), 5,5′-dithiobis-2-nitrobenzoic acid (DTNB) were purchased from Sisco Research Laboratories Pvt. Ltd. (Mumbai, India). β- Nicotinamide adenine dinucleotide phosphate reduced (NADPH)

Effect of TQ-SLNs and TQ-S on body weight mass in 3-NP induced rats

Intraperitoneal administration of 3-NP (10 mg/kg) resulted ina significant (P < 0.01) decline in the body weight of the animals as compared to vehicle treated group. Simultaneously, treatment with TQ-SLNs (10and 20 mg/kg) and TQ suspension (80 mg/kg)significantly (P < 0.01) attenuate the loss in body weight caused by 3-NP. Moreover, TQ-S at 40 mg/kg treatment also showed a significant (P < 0.05) rise in body weight measurement. Subsequently, treatment with TQ-SLNs alone and blank SLNs did not

Discussion and conclusion

Systemic administration of 3-NP has been suggested to impair energy metabolism, ROS generation and finally causes apoptosis, therefore it is considered to be a reliable tool in mimicking HD in animal models [49]. Recently, nanoparticulate system has been raised to improve the therapeutic strategy for brain oriented degenerative diseases like HD. Solid lipid nanoparticles (SLNs) physicochemical characteristics are particularly regarded to address the critical issues related to the brain

Conflicts of interest

The authors declare that no conflicts of interest exist.

Acknowledgement

The first author is grateful to UGC for the financial support in the form of JRF – UGC – Non Net Fellowship (Co/Tara/UGC-Non-Net/UGC-XII Plan/Medical Bio/2014/703 dated 28th March 2014).

References (79)

  • D.A. Shear et al.

    Comparison of intrastriatal injections of quinolinic acid and 3-nitropropionic acid for use in animal models of Huntington’s disease

    Prog. Neuro-Psychoph

    (1998)
  • K.L. Haik et al.

    Quinolinic acid released from polymeric brain implants causes behavioral and neuroanatomical alterations in a rodent model of Huntington’s disease

    Exp. Neurol.

    (2000)
  • O.H. Lowry et al.

    Protein measurement with the Folin phenol reagent

    J. Biol. Chem.

    (1951)
  • R.L. Levine et al.

    Determination of carbonyl content in oxidatively modified proteins

    Methods Enzymol.

    (1990)
  • L.C. Green et al.

    Analysis of nitrate, nitrite, and [15N] nitrate in biological fluids

    Anal. Biochem.

    (1982)
  • H. Aebi

    Catalase in vitro

    Methods Enzymol.

    (1984)
  • G.L. Ellman

    Tissue sulfhydryl groups

    Arch. Biochem. Biophys.

    (1959)
  • C.R. Wheeler et al.

    Automated assays for superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase activity

    Anal. Biochem.

    (1990)
  • I. Carlberg et al.

    Purification and characterization of the flavoenzyme glutathione reductase from rat liver

    J. Biol. Chem.

    (1975)
  • W.H. Habig et al.

    Assays for differentiation of glutathione–S–Transferases

    Meth. Enzymol.

    (1981)
  • S.T. Omaye et al.

    Selected methods for the determination of ascorbic acid in animal cells, tissues and fluids

    Methods Enzymol.

    (1979)
  • I. Tasset et al.

    Melatonin improves 3-Nitropropionic acid induced behavioral alterations and neurotrophic factors levels

    Prog. Neuro-Psychoph

    (2011)
  • P.J. Miller et al.

    3-Nitropropionic acid neurotoxicity: visualization by silver staining and implications for use as an animal model of Huntington’s disease

    Exp. Neurol.

    (1997)
  • P. Blasi et al.

    Solid lipid nanoparticles for targeted brain drug delivery

    Adv. Drug Deliv. Rev.

    (2007)
  • J.A. Saydoff et al.

    Oral uridine pro-drug PN401 decreases neurodegeneration, behavioral impairment, weight loss and mortality in the 3-nitropropionic acid mitochondrial toxin model of Huntington’s disease

    Brain Res.

    (2003)
  • W.T. Lee et al.

    Magnetic resonance imaging and spectroscopy in assessing 3-nitropropionic acid-induced brain lesions: an animal model of Huntington’s disease

    Prog. Neurobiol.

    (2004)
  • S. Jamwal et al.

    Spermidine ameliorates 3-nitropropionic acid-induced striatal toxicity: possible role of oxidative stress, neuroinflammation and neurotransmitters

    Physiol. Behav.

    (2016)
  • I. Dalle-Donne et al.

    Protein carbonyl groups as biomarkers of oxidative stress

    Clin. Chim. Acta

    (2003)
  • C.L. Powell et al.

    Expression of base excision DNA repair genes as a biomarker of oxidative DNA damage

    Cancer Lett.

    (2005)
  • M.S. Al-Ghamdi

    The anti-inflammatory, analgesic and antipyretic activity of Nigella sativa

    J. Ethnopharmacol.

    (2001)
  • R. Sandhir et al.

    Quercetin supplementation is effective in improving mitochondrial dysfunctions induced by 3-nitropropionic acid: implications in Huntington’s disease

    Biochim. Biophys. Acta

    (2013)
  • Y. Kono et al.

    Superoxide radicals inhibit catalase

    J. Biol. Chem.

    (1982)
  • Y. Kondo et al.

    Vitamin C depletion increases superoxide generation in brains of SMP30/GNL knockout mice

    Biochem. Biophys. Res. Commun.

    (2008)
  • G.V. Rebec et al.

    A vitamin as neuromodulator: ascorbate release into the extracellular fluid of the brain regulates dopaminergic and glutamatergic transmission

    Prog. Neurobiol.

    (1994)
  • K. Gopinath et al.

    Neuroprotective effect of naringin, a dietary flavonoid against 3-Nitropropionic acid-induced neuronal apoptosis

    Neurochem. Int.

    (2011)
  • B. Shukitt-Hale et al.

    Spatial learning and memory deflcitsinduced by dopamine administration with decreased glutathione

    Free Radic. Biol. Med.

    (1998)
  • A. Hariharan et al.

    Potential of protease inhibitor in 3-nitropropionic acid induced Huntington’s disease like symptoms: mitochondrial dysfunction and neurodegeneration

    Neurotoxicology

    (2014)
  • S.E. Browne et al.

    The energetics of Huntington’s disease

    Neurochem. Res.

    (2004)
  • S. Ramaswamy et al.

    Animal models of Huntington’s disease

    ILAR J.

    (2007)
  • Cited by (0)

    View full text