Effects of quercetin loaded nanostructured lipid carriers on the paraquat-induced toxicity in human lymphocytes

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Highlights

  • Quercetin loaded NLC protect lymphocytes in paraquat toxicity.

  • Quercetin loaded NLC mitigate paraquat-medaited oxidative damage.

  • Paraquat-mediated mitochondrial toxicity reduced by quercetin loaded NLC.

Abstract

Paraquat (PQ) as a herbicide and an environmental pollutant with increasing importance due to its toxicity to humans and animals. This study aimed to evaluate the protective and antioxidant activity of quercetin loaded Nanostructured Lipid Carriers (QNLC) against toxicity induced by PQ. Blood lymphocytes were prepared using Ficoll polysaccharide and subsequently by gradient centrifugation. The QNLC was prepared using an ultra-sonication method, which was characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM).

The viability, reactive oxygen species (ROS), lipid peroxidation (LPO), mitochondrial membrane potential (MMP), lysosome membrane integrity, Bax and Bcl2 gene expression were evaluated in human isolated lymphocytes.

The results showed spherical QNLCs with nano-size range (52.7 nm) and high drug encapsulation efficiency (98.5% -96%). The results also indicated that PQ induced cell death, as well as ROS production, decreased by QNLC in human lymphocytes. Also, QNLC meaningfully restored MMP reduction, lysosomal membrane destabilization, and lipid peroxidation and were capable of preventing PQ-treated change in Bax and Bcl2 gene expression. We report that QNLC, have a significantly higher capacity to prevent PQ-induced toxicity than Q itself. It is suggested that the QNLC is a promising antioxidant for drug delivery to be used as a therapeutic and prophylactic agent for PQ poisoning.

Introduction

Exposure to environmental chemicals has been considered as a widespread health problem worldwide (Ruiz-Guzmán et al., 2017). Accidental or intentional poising by means of several pesticides or herbicides is a serious issue in the field of toxicology. Paraquat (N, N′-dimethyl-4,4′-bipyridinium dichloride; PQ) as a common herbicide worldwide (Yao et al., 2015), has several limitations in the context of marketing and application. However, acute intoxication and/or long-term accretion of PQ might result in organ injury with a mortality rate of over 60–70% (Seok et al., 2009). The absorption of PQ through the gastrointestinal tract occurs within 2 h subsequent to digestion. Then, the poison is accumulated in different body organs such as liver, kidney, and lung as well as the central nervous system (Gil et al., 2014).

The function of the immune system can be disturbed by different chemicals, including pesticides (Corsini et al., 2013). The enhanced immunomodulation has been observed after PQ treatment in several in vitro studies (Paolillo et al., 2011). Absolute lymphocyte count has been indicated as an independent marker of prognosis for patients with follicular lymphomas and diffuses large B-cell lymphoma (Behl et al., 2007; Kim et al., 2007). Although the comprehensive mechanism of PQ cytotoxicity remains obscure, interfering with intracellular electron transferring processes and induction of oxidative stress has been recognized to play key roles (Dinis-Oliveira et al., 2008). Accordingly, a great deal of work has been done to introduce effective antioxidant compounds to combat PQ-induced toxicity. Vitamins, N-acetylcysteine (NAC), glutathione, melatonin, β-carotene and the enzyme superoxide dismutase have been utilized in this context (Suntres, 2002; Yeh et al., 2006). However, the uses of antioxidants or immunosuppressants have several restrictions (Gawarammana and Buckley, 2011). Antioxidants agents derived from natural fruits and vegetables have been shown to support the function of endogenous antioxidant defense. Quercetin (Q) as a flavonoid antioxidant is one of the main members of these exogenous compounds. In spite of potent antioxidant properties, the clinical application of Q is limited due to some drawbacks such as low solubility and bioavailability (Justino et al., 2004). Novel drug delivery systems have, in particular, the application of nanotechnology has offered new beneficial pharmacological effects of naturally occurring flavonoids.

Nanostructured Lipid Nanocarriers (NLC) as second-generation lipid carriers developed for the delivery of lipophilic drugs and marketed for therapeutic and cosmetic uses. Drug delivery through the NLC application is advantageous over other methods because of its similarity to biomembranes structure, biocompatibility and biodegradability, and safety (Salvi and Pawar, 2019). In addition, these carriers could be modified and used as diagnostic tools(Salvi and Pawar, 2019). Application of Q in novel nanoformulations is deemed promising in reducing drug/toxin side effects. While nanocrystalline forms of Q are considered as simple and cost-effective techniques, but instability and the need for organic solvents are disadvantages. QNLC successfully enhances the solubility of Q as a lipophilic agent (Kakran et al., 2012a).

Thus, the present study aims to establish a novel therapeutic agent via the synthesis of an NLC of Q to manage the PQ-induced toxicity. So, the plausible protective role of QNLC against PQ-induced toxicity in peripheral blood lymphocytes and the function of subcellular organelles and the genes related to the apoptotic machinery were evaluated.

Section snippets

Materials and methods

Quercetin was achieved from Merck Company (Darmstadt, Germany). Paraquat (PQ) (methyl viologen dichloride hydrate, CAS number 75365–73-0, 98% purity) was procured from Sigma Aldrich (St. Louis, USA). Tween 20, Tween 80, Compritol 888 (glyceryl dibehenate), Precirol ATO5 (glyceryl palmitostearate), Poloxamer 188, Sodium lauryl sulfate (SLS) and Dioctyl sodium Sulfosuccinate (DOSS) were obtained from Sigma–Aldrich Company (Munich, Germany). All other materials were of analytical grade.

Characterization of quercetin-loaded NLCs

The morphology for the prepared QNLC was imaged using transmission electron microscopy (TEM). The obtained results for TEM image presented spherical particles with nanometer size (Fig. 1A). The mean particle size resulted from the TEM method was about 40 nm which was somewhat lower than the mean particle size gained from the DLS method (52.7 nm)(Fig. 1B). The prepared QNLCs also showed an encapsulation efficiency of 98.5% -96%.

Biochemical test results

Our results showed that PQ substantially diminished the viability of

Discussion

The immune system has a critical role in the maintenance of human health and is affected by the toxic effects of widespread chemicals. Needless to say, suppression of the immune system will result in enhanced incidence and severity of infectious disorders as well as tumorigenesis (Galluzzi et al., 2017). Immunotoxic substances can disturb the function of the immune system by both the suppression and enhancement of its activity (Mokarizadeh et al., 2015).

Plant's photosystems are sensitive to

Funding

The authors would like to acknowledge the financial support of Maragheh University of Medical Sciences for this research under grant number IR.MARAGHEHPHC.REC.1396.24.

Declaration of Competing Interest

None.

Acknowledgment

The authors are thankful to Maragheh University of Medical Sciences, Maragheh, Iran.

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