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Evaluation of the Effects of Quercetin on Brain Lesions Secondary to Experimental Hydrocephalus in Rats

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Evaluation of the Effects of Quercetin on Brain Lesions Secondary to Experimental Hydrocephalus in Rats Child's Nervous System (2019) 35:2299–2306 https://doi.org/10.1007/s00381-019-04184-2 ORIGINAL ARTICLE Evaluation of the effects of quercetin on brain lesions secondary to experimental hydrocephalus in rats G. B. Sampaio1 & S. C. Da Silva2 & T. H. Romeiro2 & P. D. S. Beggiora2 & H. R. Machado2 & L. D. S. Lopes2 Received: 31 March 2019 /Accepted: 28 April 2019 /Published online: 27 May 2019 # Springer-Verlag GmbH Germany, part of Springer Nature 2019 Abstract Introduction Hydrocephaly is a disease that affects not only the dynamics of the cerebrospinal fluid, but also other structures of the central nervous system. Although shunt is effective in reducing ventriculomegaly, many neurological damages are not reversed with surgery. Several studies demonstrate that oxidative stress is involved in the genesis of hydrocephalus lesions. Objective Evaluate the neuroprotective response of quercetin in hydrocephalus. Materials and methods Male newborns rats were used, which received the 15% kaolin injection in the cisterna magna for induction of hydrocephalus. They were divided into control group (C), untreated hydrocephalic (HN), shunted hydrocephalic (HD), hydrocephalic treated with distilled water (HA), hydrocephalic treated with distilled water and shunt (HDA), hydroce- phalic treated with quercetin peritoneal (HQp), hydrocephalic treated with quercetin peritoneal and shunt (HDQp), hydrocephalic treated with quercetin by gavage (HQg), and hydrocephalus treated with quercetin by gavage and shunt (HDQg). Results Quercetin significantly improved the immunohistochemical markers, mainly caspase and GFAP. There were no signif- icant changes in clinical/behavioral assessment. The use of isolated quercetin does not alter the volume and ventricular size, and the realization of ventriculo-subcutaneous shunt in newborn rats with hydrocephalus presents a high morbi-mortality. Conclusion The use of quercetin shows laboratory improvement of the effects of glial lesion and corpus callosum fibers and is therefore not justified by the use of the routine substance as neuroprotective. Keywords Hydrocephalus . Neuroprotective drugs . Oxidative stress . Shunt Introduction [3]. Another injured structure is the white matter, which in later cases of ventricular dilation, may also lose myelin. Hydrocephaly may be defined as an increase in the amount of A significant and important advance in the history of the cerebrospinal fluid (CSF) in the ventricular or subarachnoid treatment of hydrocephalus was the introduction of unidirec- spaces, generating intracranial hypertension. The imbalance tional shunt systems, with the objective of retrieving excess between the production and absorption of CSF leads to ven- fluid from the cerebral ventricles to other body cavities, can- tricular dilatation and consequent distortions in the cerebral celing the pathophysiological basis of intracranial hyperten- parenchyma, compressing the hemispheres against the inner sion. This advance promoted a marked reduction in mortality surface of the skull [2]. The first affected structure is the ven- and morbidity in children with hydrocephalus [9]. tricular parenchyma, which undergoes compression and Oxidative stress (EO) occurs in hydrocephalus, due to stretching; being destroyed in isolated or even totally ruptured the aggression caused to the cerebral parenchyma by the forces of compression, stretching and consequently ische- mia of the brain tissue. This action leads to tissue damage, with the production of free radicals, predominantly through * G. B. Sampaio [email protected] lipid peroxidation, which has a toxic effect on the walls of cerebral arterial vessels. EO is a biological condition in which the imbalance between the production of reactive 1 Hospital da Criança e Maternidade, Av. Jamil Feres Kfouri, 60 - Jardim Panorama, São José do Rio Preto, SP 15091-240, Brazil oxygen species (ROS) and their detoxification occurs through biological systems that remove and/or repair them. 2 Developmental neuropathology laboratory - Faculdade de Medicina da Universidade de São Paulo, Campus Ribeirão Preto - SP, Av However, all organisms have an intracellular environment bandeirantes, 3900, Ribeirão Preto, São Paulo 14040-900, Brazil of reductive nature, so that there is a balance between the 2300 Childs Nerv Syst (2019) 35:2299–2306 oxidized and reduced molecules, thus keeping the enzymes lesion. Quercetin acts as a neuroprotectant directly in the at the expense of metabolic energy. intracellular mechanisms of the ischemic cascade, Quercetin is a natural flavonoid that has pharmaco- aiming at the rescue of the area of hypoperfusion, still logical properties, such as anti-inflammatory, anticarci- viable, surrounding the infarct. Therefore, the study of nogenic, antiviral, antihistamines, and antioxidants. It the neuroprotective effects of quercetin in hydrocephalus has protective cardiovascular activity, reducing the risk is extremely important and may represent a new treat- of death due to coronary diseases and reducing the in- ment for the disease. cidence of myocardial infarction. [13]. Among its main actions, it can remove free radicals, exerting a cytoprotective role in situations of risk of cellular Material and methods damage. Quercetin has been shown to inhibit the in vitro ox- idation of low density lipoprotein (LDL) by macrophages and All procedures involving the animals are in accordance to reduce the cytotoxicity of oxidized LDL [7]. with the guidelines established by the Brazilian College It is the most abundant flavonoid present in the human diet, of Animal Experimentation (COBEA) and approved by representing about 95% of the total flavonoids ingested [13]. the local committee held by the Ethics Committee on Their estimated daily intake ranges from 50 mg to 500 mg Animal Experimentation (CETEA)—University of São daily [4]. It is present in vegetables and fruits. Being found in Paulo at Ribeirão Preto Medical School (FMRP-USP), high concentrations in apples, onions, tea, red wine, and broc- Protocol No. 32/2016. All efforts were made to mini- coli. It is extracted very easily, because it is in great quantity. mize the suffering and the number of animals used in The protective effect of quercetin was superior to that this research. of vitamin C, by preventing the effect of reducing glu- At 7 days of age, the animals were randomly selected tathione, and protecting the brain from oxidative stress and submitted to hydrocephalus induction by induced by neurotoxicity due to its structural properties intracisternal injection of kaolin or maintained without and physiological benefits [7]. Quercetin inhibits the intervention. The space between the posterior margin of enzymes cyclooxygenase and lipoxygenase, which re- the foramen magnum in the occipital bone and the cranial duces the production of the main inflammatory media- border of the dorsal arch of the first cervical vertebra was tors: prostaglandins and leukotrienes. It inhibits hista- identified. With a short bevel Mise 0.3 dental needle, a mine production, stabilizing basophils and mast cells. suboccipital puncture and slow percutaneous injection of Quercetin inhibits the process of free radical formation 0.04 ml of a 15% kaolin (Merck®) suspension in sterile in three different steps, initiation (by interaction with autoclaved distilled water was performed. superoxide ions), formation of hydroxyl radicals (by At 7 days after induction of hydrocephalus with kaolin, the chelating iron ions), and lipid peroxidation (by reacting animals were anesthetized with 10% Isoflurane with lipid peroxy radicals). (BioChimo®) with continuous oxygen therapy, followed Many studies report that flavonoids in their free or by administration of Cephalotin 50 mg/kg for preopera- glycosylated form are absorbed in the gastrointestinal tive infectious prophylaxis, degermation of the head, tract and metabolized to glucuronidate or conjugated sul- neck, and trunk with deodorant iodine and painted with fate. These metabolites circulate in the blood being ex- alcoholic iodine. A cut was performed in the cranial re- creted in bile and urine. Quercetin is completely convert- gion near the right coronal suture and a trepanation for- ed into methylated conjugates in plasma, after administra- ward of the right coronal suture using a drill and a rotary tion, in both rats and humans. Quercetin is absorbed in the punch; the dorsal region of the trunk of the animal was intestinal microflora and excreted in bile and urine as dissected using a dissector. Next, a ventricular punction glucuronidate and conjugated sulfate within 48 h. was performed using a ventricular catheter. Using a Subsequently, it is degraded by intestinal bacteria in phe- prolene wire 5.0, the proximal catheter was attached to nolic acid, 3-hydroxyphenylacetic acid and 3,4- the bone. After reviewing the surgical technique, the pro- dihydroxyphenylacetic acid within ring B. cedure was performed the closure of the skin in a single As cerebral permeability is controlled by psycho-chemical plane with prolene 5.0 (Fig. 1). The technique was devel- characteristics such as hydrophobicity or lipophilicity, querce- oped in the research laboratory, detailing the procedures tin may enter brain regions benefiting from antioxidant and recently published [16](Figs.2 and 3). biological functions, protecting against hydrogen peroxide- induced cytotoxicity. [4]. Experimental groups Neuroprotection is an intervention, not necessarily pharma- cological, whose objective is to avoid damage to the neural Control
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