Valproic Acid Sodium Research Articles

In Rat Organotypic Hippocampal Slice Cultures, Conventional Antiepileptic Medications are Unable to Inhibit Epileptiform Activity

Context and Goal: Prior research has shown that organotypic hippocampal slice cultures (OHSCs) can elicit tonic-clinic seizure-like events (SLEs), which resemble the electroencephalographic correlates of limbic seizures in humans and animals. We have investigated whether OHSCs can be used as in vitro models of limbic seizures to research seizures processes and vetting novel antiepileptic substances. Experimental Strategy: The interface method was used to cultivate OHSCs. Under submerged conditions, the levels of extracellular potassium and neuronal activity were observed. Lowering magnesium concentrations or administering the potassium channel blocker 4-aminopyridine both caused SLEs. Analysis was done on the impact of common antiepileptic medications (AEDs) on SLEs, including carbamazepine, phenytoin, valproic acid, clonazepam, diazepam, and phenobarbital sodium. Important Outcomes: AEDs did not stop the over 93% of OHSCs from happening induction of SLEs or cease current seizure activity even in the presence of hazardous dosages. The postnatal age at explanation (P2–P10), the manner of seizure provocation, and the length of in vitro cultivation (2 months) had no bearing on this chemoresistance. GABAA-agonist muscimol or glutamate antagonists were able to reversibly block SLEs. Inferences and Conclusions: Unlike animal models of thermoresistant seizures, where responders and nonresponses may only be distinguished after an experiment, we describe an easy to set up in vitro model of tonic-clinic SLEs that is a priori thermoresistant. OHSCs may be useful for investigating the mechanisms underlying medication-resistant seizures and for the discovery of novel anticonvulsive substances that may one day be used to treat drugresistant epilepsy.

Valproic acid treatment attenuates cisplatin-induced kidney injury by suppressing proximal tubular cell damage.

Cisplatin treatment is effective against several types of carcinomas. However, it frequently leads to kidney injury, which warrants effective prevention methods. Sodium valproic acid is a prophylactic drug candidate with a high potential for clinical application against cisplatin-induced kidney injury. Therefore, in this study, we aimed to elucidate the mechanism underlying the prophylactic effect of valproic acid on cisplatin-induced kidney injury in a mouse model and HK2 and PODO cells with cisplatin-induced toxicity. In the mouse model of cisplatin-induced kidney injury, various renal function parameters and tubular damage scores were worsened by cisplatin, but they were significantly improved upon combination with valproic acid. No difference was observed in cisplatin accumulation between the cisplatin-treated and valproic acid-treated groups in whole blood and the kidneys. The mRNA expression levels of proximal tubular damage markers, apoptosis markers, and inflammatory cytokines significantly increased in the cisplatin group 72 h after cisplatin administration but significantly decreased upon combination with valproic acid. In HK2 cells, a human proximal tubular cell line, the cisplatin-induced decrease in cell viability was significantly suppressed by co-treatment with valproic acid. Valproic acid may inhibit cisplatin-induced kidney injury by suppressing apoptosis, inflammatory responses, and glomerular damage throughout the kidneys by suppressing proximal tubular cell damage. However, prospective controlled trials need to evaluate these findings before their practical application.

GABAB1 receptor knockdown in prefrontal cortex induces behavioral aberrations associated with autism spectrum disorder in mice

Autism spectrum disorder (ASD) is a set of heterogeneous neurodevelopmental disorders, characterized by social interaction deficit, stereotyped or repetitive behaviors. Apart from these core symptoms, a great number of individuals with ASD exhibit higher levels of anxiety and memory deficits. Previous studies demonstrate pronounced decrease of γ-aminobutyric acid B1 receptor (GABAB1R) protein level of frontal lobe in both ASD patients and animal models. The aim of the present study was to determine the role of GABAB1R in ASD-related behavioral aberrations. Herein, the protein and mRNA levels of GABAB1R in the prefrontal cortex (PFC) of sodium valproic acid (VPA)-induced mouse ASD model were determined by Western blot and qRT-PCR analysis, respectively. Moreover, the behavioral abnormalities in naive mice with GABAB1R knockdown mediated by recombinant adeno-associated virus (rAAV) were assessed in a comprehensive test battery consisted of social interaction, marble burying, self-grooming, open-field, Y-maze and novel object recognition tests. Furthermore, the action potential changes induced by GABAB1R deficiency were examined in neurons within the PFC of mouse. The results show that the mRNA and protein levels of GABAB1R in the PFC of prenatal VPA-induced mouse ASD model were decreased. Concomitantly, naive mice with GABAB1R knockdown exhibited ASD-like behaviors, such as impaired social interaction and communication, elevated stereotypes, anxiety and memory deficits. Patch-clamp recordings also revealed that GABAB1R knockdown provoked enhanced neuronal excitability by increasing action potential discharge frequencies. Overall, these findings support a notion that GABAB1R deficiency might contribute to ASD-like phenotypes, with the pathogenesis most likely resulting from enhanced neuronal excitability. SubheadingsGABAB1 Knockdown Induces Behavioral Aberrations with ASD

Scalp acupuncture improves disordered behavior in the valproic acid-induced rat model of autism: 头针改善丙戊酸钠诱导的孤独症大鼠行为学及其机制研究

ObjectivesTo investigate the underlying mechanisms of scalp acupuncture treatment (SAT) on autism spectrum disorder (ASD). MethodsThirty male Wistar rat pups that had been prenatally exposed to valproic acid sodium (VPA) were randomly divided into the VPA, VPA+acupoint, and VPA+ non-acupoint groups using the random number table method, with 10 rats in each group. Ten pups who had been prenatally exposed to saline were assigned to the control group (CG). There was no intervention in either the control or VPA groups. In the acupoint group, “Shenting (GV24),” bilateral “Benshen (GB13)” were manipulated. In the non-acupoint group, the area below the costal space was stimulated. Acupuncture stimulation lasted for 40 min, with manual twisting of the needles every 10 min, 5 days/week, with 2 days of rest per week, for a total duration of 4 weeks. After the corresponding treatments, behavioral tests (including the open field, social interaction, and Morris water maze tests) were performed to evaluate the therapeutic effects. RT-PCR and Western blotting were performed to detect the expression of neuregulin 1 (NRG1)/ErbB4. ResultsIn the open field test, the activity time spent in the central area in the VPA+acupoint group was significantly longer than that in the VPA group and VPA+ non-acupoint group (both P<0.05). The total length in the VPA+acupoint group was significantly longer than that in the VPA group (P<0.05). The number of bouts in the central area of the VPA+acupoint group was significantly higher than that of the VPA group (P<0.05). In session I of social interaction test, all experimental rats spent more time interacting with stranger 1 (all P<0.05). In session II, the CG and VPA+acupoint groups rats showed more interest in searching for new strangers, but the VPA+non-acupoint group spent more time interacting with stranger 1 than with stranger 2(all P<0.05). In the Morris water maze test, compared with the VPA group, the latency of the VPA+acupoint group was shorter (day 2, 3, 4, 5, P<0.05); compared with VPA+acupoint group, the latency of the VPA+non-acupoint group was longer (day 2, 4, P<0.05). The mean distance in the VPA+acupoint group was shorter than that in the VPA group (day 3, 5, P<0.05). The platform quadrant time of the VPA+non-acupoint group was significantly shorter than that of the VPA+acupoint group (P<0.05) (day 6). The VPA+acupoint group had more platform crossings than the VPA group (P<0.05), and the VPA+ non-acupoint group had fewer platform crossings than the VPA+acupoint group (P<0.05) (day 6). After SAT, the expression levels of NRG1 and ErbB4 proteins in the VPA+acupoint group were significantly increased than those in the VPA group (both P<0.05) in the medial prefrontal cortex (mPFC). The expression levels of NRG1 and ErbB4 proteins in VPA+non-acupoint group were significantly less than those in the VPA+acupoint group (both P<0.05) in the mPFC. After SAT, the expression levels of NRG1 and ErbB4 proteins in the VPA+acupoint group were significantly higher than those in the VPA group (both P<0.05) in the hippocampus. The expression levels of NRG1 and ErbB4 proteins in the VPA+non-acupoint group were significantly lower than those in the VPA+acupoint group (both P<0.05) in the hippocampus. After SAT, the expression levels of mRNA levels of NRG1-ErbB4 in the mPFC and hippocampus in VPA+acupoint group were significantly higher than those in the VPA group (all P<0.05). The mRNA expression levels of NRG1-ErbB4 in the mPFC and hippocampus in VPA+ non-acupoint group were significantly lower than those in the VPA+acupoint group (all P<0.05). ConclusionsOur results suggest that SAT can improve ASD-like behaviors in young rats in a VPA model of ASD. This may be related to its function in upregulating the expression of protein and gene of NRG1 and its receptor ErbB4 in the mPFC.

Study on the protective effect of sodium valproic acid on carbonyl cyanide 3-chlorophenylhydrazone-induced oxidative stress injury in osteoblasts

To explore the protective effects of sodium valproic acid (VPA) on oxidative stress injury of osteoblasts induced by carbonyl cyanide 3-chlorophenylhydrazone (CCCP) and its mechanism. Osteoblasts were isolated from the skulls of 10 newborn Sprague Dawley rats and cultured by tissue block method, and the 1st generation cells were identified by alkaline phosphatase (ALP) and alizarin red staining. The 3rd generation osteoblasts were cultured with 2-18 μmol/L CCCP for 2-18 minutes, and cell counting kit 8 (CCK-8) was used to detect the cell survival rate. An appropriate inhibitory concentration and culture time were selected for the preparation of osteoblasts oxidative stress injury model based on half maximal concentration principle. The cells were cultured with 0.2- 2.0 mmol/mL VPA for 12-72 hours, and CCK-8 was used to detect cell activity, and appropriate concentration was selected for further treatment. The 3rd generation cells were randomly divided into 4 groups, including blank control group (normal cultured cells), CCCP group (the cells were cultured according to the selected appropriate CCCP concentration and culture time), VPA+CCCP group (the cells were pretreated according to the appropriate VAP concentration and culture time, and then cultured with CCCP), VPA+CCCP+ML385 group (the cells were pretreated with 10 μmol/L Nrf inhibitor ML385 for 2 hours before VPA treatment, and other treatments were the same as VPA+CCCP group). After the above treatment was complete, the cells of 4 groups were taken to detect oxidative stress indicators [reactive oxygen species (ROS), superoxide dismutase (SOD), malondialdehyde (MDA)], cell apoptosis rate, ALP/alizarin red staining, and the relative expressions of osteogenic related proteins [bone morphogenetic protein 2 (BMP-2), RUNX2], anti-apoptotic family protein (Bcl2), apoptotic core protein (Cleaved-Caspase-3, Bax), channel protein (Nrf2) by Western blot. The osteoblasts were successfully extracted. According to the results of CCK-8 assay, the oxidative stress injury model was established by 10 μmol/L CCCP cultured for 10 minutes and 0.8 mmol/mL VPA cultured for 24 hours was selected for subsequent experiments. Compared with blank control group, the activity and mineralization capacity of osteoblasts in CCCP group decreased, the contents of ROS and MDA increased, the activity of SOD decreased, and the apoptosis rate increased. Meanwhile, the relative expressions of BMP-2, RUNX2, and Bcl2 decreased, and the relative expressions of Cleaved-Caspase-3, Nrf2, and Bax increased. The differences were significant ( P<0.05). After further VPA treatment, the oxidative stress damage of osteoblasts in VPA+CCCP group was relieved, and the above indexes showed a recovery trend ( P<0.05). In VPA+CCCP+ML385 group, the above indexes showed an opposite trend ( P<0.05), and the protective effects of VPA were reversed. VPA can inhibit the CCCP-induced oxidative stress injury of osteoblasts and promote osteogenesis via Keap1/Nrf2/Are pathway.

Neuroprotective Effect of Celastrus Paniculatus Seed Extract on Epilepsy and Epilepsy-associated Cognitive Deficits.

Cognitive deficit is one of the common comorbidity accompanying epilepsy. The present study evaluated the effect of Celastrus paniculatus seed extract on seizure severity and cognitive deficit following the pentylenetetrazole (PTZ)-induced chemical kindling model. PTZ kindling model was developed by daily administration of the sub-convulsive dose of PTZ 30 mg/kg for four weeks. After four weeks of induction, the following treatment, namely sodium valproic acid (SVA) 200 mg/kg, C. paniculatus 500 mg\kg, pergolide 2 mg/kg, C. paniculatus (250 mg\kg)+ Pergolide (1 mg/kg), and C. paniculatus (250 mg\kg)+ SVA (100 mg/kg) were administered 30 minutes prior to PTZ (30 mg/kg) injection for a period of next 14 days. Neurobehavioral parameters, including superoxide dismutase (SOD), Catalase (CAT), glutathione (GSH), and dopamine levels were assessed and the Morris water maze test (MWM) and Grip strength test (GPS) were performed. Hematoxylin & Eosin (H&E) staining of hippocampal cornu ammonis (CA1), CA2, CA3, dentate gyrus (DG), and frontal cortex was performed. C. paniculatus (500 mg/kg) alone and in combination (C. paniculatus (250 mg\ kg)+ pergolide (1 mg/kg) and C. paniculatus (250 mg\kg)+ SVA (100 mg/kg)) significantly (P<0.05) reduced the seizure score, mean latency time, and distance traveled in the MWM. However, no significant effect was seen in GPS. Biochemical analysis showed elevated antioxidant markers, namely GSH, CAT, and SOD, and also elevated dopamine levels. C. paniculatus and its combination also significantly (P<0.05) protected against neuronal loss in the hippocampus and frontal cortex evidenced by H&E staining. C. paniculatus alone and in combination with other agents may have the potential to treat epilepsy and associated cognitive deficits.

Inhibiting silence information regulator 2 and glutaminase in the amygdala can improve social behavior in autistic rats.

To investigate the underlying molecular mechanisms by which silence information regulator (SIRT) 2 and glutaminase (GLS) in the amygdala regulate social behaviors in autistic rats. Rat models of autism were established by maternal sodium valproic acid (VPA) exposure in wild-type rats and SIRT2-knockout ( SIRT2 -/-) rats. Glutamate (Glu) content, brain weight, and expression levels of SIRT2, GLS proteins and apoptosis-associated proteins in rat amygdala at different developmental stages were examined, and the social behaviors of VPA rats were assessed by a three-chamber test. Then, lentiviral overexpression or interference vectors of GLS were injected into the amygdala of VPA rats. Brain weight, Glu content and expression level of GLS protein were measured, and the social behaviors assessed. Brain weight, amygdala Glu content and the levels of SIRT2, GLS protein and pro-apoptotic protein caspase-3 in the amygdala were increased in VPA rats, while the level of anti-apoptotic protein Bcl-2 was decreased (all P<0.01). Compared with the wild-type rats, SIRT2 -/- rats displayed decreased expression of SIRT2 and GLS proteins in the amygdala, reduced Glu content, and improved social dysfunction (all P<0.01). Overexpression of GLS increased brain weight and Glu content, and aggravated social dysfunction in VPA rats (all P<0.01). Knockdown of GLS decreased brain weight and Glu content, and improved social dysfunction in VPA rats (all P<0.01). The glutamate circulatory system in the amygdala of VPA induced autistic rats is abnormal. This is associated with the upregulation of SIRT2 expression and its induced increase of GLS production; knocking out SIRT2 gene or inhibiting the expression of GLS is helpful in maintaining the balanced glutamate cycle and in improving the social behavior disorder of rats.

Effect of stigma maydis polysaccharide on the gut microbiota and transcriptome of VPA induced autism model rats.

Stigma maydis polysaccharide (SMPS) is a plant polysaccharide that participates in immune regulation and gastrointestinal motility. Autism spectrum disorder (ASD) refers to a group of neurodevelopmental disorders, and ASD patients often present intestinal microflora imbalance problems; however, there is no effective treatment method. This study explores the effect of SMPS intervention on the gut microbiota in autism model rats as well as the potential action pathways. Female Wistar rats were intraperitoneally injected with sodium valproic acid (VPA) or normal saline at embryonic day 12.5 to establish an autism model or normal control in their offspring. The offspring prenatally exposed to VPA were randomly assigned to the VPA and the SMPS groups. The SMPS group was administered SMPS from E0.5 to postnatal day (PND) 21. We performed 16S rRNA and transcriptomics analyses to reveal the gut microbiota (GM) and differentially expressed genes in the autism model rats in response to SMPS intervention. SMPS intervention significantly improved the diversity and structure of the GM in autism model rats compared with the VPA rats. Moreover, the relative abundance of Prevotellaceae and Lachnospiraceae_NK4A136_group was increased after SMPS intervention. Transcriptome sequencing showed that 496 differentially expressed genes (DEGs) were identified after SMPS administration compared with the VPA group. Meanwhile, gene ontology (GO) enrichment analysis of DEGs was showed that the SMPS group had significant 653 GO terms. SMPS intervention had a major influence on oxidative phosphorylation, retrograde endocannabinoid signaling, thermogenesis, ribosome, protein digestion and absorption, renin-angiotensin system, calcium signaling pathway, glycosphingolipid biosynthesis-ganglio series, and propanoate metabolism pathways. Overall, this study suggests that SMPS interventions in early life may have an impact on gut microbiota, and then affect the transcriptomics levels of the hippocampal tissue in the VPA-induced autism model rats. It provides scientific evidence for the role of the microbe-gut-brain axis in ASD research.

Evaluation of mitochondrial oxidative toxicity in mammalian cardiomyocytes by determining the highly reproducible and reliable increase in mitochondrial superoxides after exposure to therapeutic drugs

Mitochondria are important cytoplasmic elements present in eukaryotic cells, and are involved in converting energy to ATP through oxidative phosphorylation. Mitochondria are vulnerable to reactive oxygen species (ROS), thereby making it imperative to evaluate the toxicity. However, existing methods that evaluate mitochondrial toxicity in cardiomyocytes are limited. In the current study, we aimed to determine a mitochondrial biomarker that measures the toxicity of mitochondria, and subsequently suggest an efficient evaluation system for evaluating mitochondrial-specific oxidative toxicity. To achieve this, AC16 human cardiomyocytes, H9C2 rat cardiomyocytes were exposed to acetaminophen (AP), amiodarone hydrochloride (AMD), doxorubicin hydrochloride (Dox), valproic acid sodium salt (Val), and (Z)-4-hydroxytamoxifen (4-OHT). Mitochondrial oxidative stress was determined by staining the drug-treated cells with MitoSOX™ red fluorescence dye, followed by imaging with a fluorescence microscope. All working concentrations of Dox showed increased levels of red fluorescence in AC16 and H9C2 cells, whereas exposure to Val did not alter the red fluorescence level of both cells. Considering our results, increased MitoSOX™ subsequent to drug exposure is a highly reproducible and reliable method to measure the mitochondrial-specific oxidative toxicity. These results indicate that a screening system using MitoSOX™ has the potential to be applied as a reliable biomarker for determining mitochondrial oxidative toxicity in new drug development.

Role of valproic acid and allopurinol in neuroprotection after cardiac surgery on Bangladeshi patients

Background: Neurological complications are very much common after cardiac surgery in postoperative setting in Bangladesh. However, we are dealing these complications in our day-to-day practice and it costs money, longer hospital stays and admissions to rehabilitation facilities. The aim of the study was to evaluate the effectiveness of sodium valproic acid and allopurinol in preventing neurological injury following cardiac surgery with cardiopulmonary bypass.Methods: This cross-sectional study was conducted by the department of cardiac surgery, Bangabandhu Sheikh Mujib medical university (BSMMU), Dhaka, Bangladesh from January 2019 to June 2021. Total 70 patients were included and divided into two groups (36 patients with sodium valproic acid and allopurinol in group A and 34 patients without sodium valproic acid and allopurinol in group B). 10 patients were dropped out. As these two drugs were FDA approved was used per-orally for 5 perioperative days (2 days prior surgery, 1 day in operation day, 2 days postoperatively). Collected data were entered, checked and edited (to remove the outliers) with the help of the statistical package for social sciences (SPSS) software, version 26 and analyzed.Results: The intervention group (group A) had maximum survival rate (no mortality) less ICU stay, less hospital stay, early intubation, no neurological injury (p&lt;0.05). On the contrary group B (no intervention) had 6 mortalities, several mild to moderate neurological injury, more ICU and hospital stay and morbidity.Conclusions: Sodium valproic acid and allopurinol gives neuroprotection in on-pump cardiac surgical patients. So, these two medicines can be prescribed prophylactically on individual basis.

Phenotypic overlap between atopic dermatitis and autism

BackgroundAutism, a childhood behavioral disorder, belongs to a large suite of diseases, collectively referred to as autism spectrum disorders (ASD). Though multifactorial in etiology, approximately 10% of ASD are associated with atopic dermatitis (AD). Moreover, ASD prevalence increases further as AD severity worsens, though these disorders share no common causative mutations. We assessed here the link between these two disorders in the standard, valproic acid mouse model of ASD. In prior studies, there was no evidence of skin involvement, but we hypothesized that cutaneous involvement could be detected in experiments conducted in BALB/c mice. BALB/c is an albino, laboratory-bred strain of the house mouse and is among the most widely used inbred strains used in animal experimentation.MethodsWe performed our studies in valproic acid (VPA)-treated BALB/c hairless mice, a standard mouse model of ASD. Mid-trimester pregnant mice received a single intraperitoneal injection of either valproic acid sodium salt dissolved in saline or saline alone on embryonic day 12.5 and were housed individually until postnatal day 21. Only the brain and epidermis appeared to be affected, while other tissues remain unchanged. At various postnatal time points, brain, skin and blood samples were obtained for histology and for quantitation of tissue sphingolipid content and cytokine levels.ResultsAD-like changes in ceramide content occurred by day one postpartum in both VPA-treated mouse skin and brain. The temporal co-emergence of AD and ASD, and the AD phenotype-dependent increase in ASD prevalence correlated with early appearance of cytokine markers (i.e., interleukin [IL]-4, 5, and 13), as well as mast cells in skin and brain. The high levels of interferon (IFN)γ not only in skin, but also in brain likely account for a significant decline in esterified very-long-chain N-acyl fatty acids in brain ceramides, again mimicking known IFNγ-induced changes in AD.ConclusionBaseline involvement of both AD and ASD could reflect concurrent neuro- and epidermal toxicity, possibly because both epidermis and neural tissues originate from the embryonic neuroectoderm. These studies illuminate the shared susceptibility of the brain and epidermis to a known neurotoxin, suggesting that the atopic diathesis could be extended to include ASD.

Влияние афобазола на изменения в раннем постнатальном периоде у мышей линии BALB/c с фетальным вальпроатным синдромом

Введение. Воздействие вальпроевой кислотой во время беременности у грызунов широко используется для моделирования расстройств аутистического спектра (РАС). Цель исследования - изучение ранних поведенческих изменений у мышей BALB/c, пренатально подвергшихся однократному воздействию натриевой соли вальпроевой кислоты (400 мг/кг), и возможности их коррекции афобазолом. Методика. Объект исследования - мыши линии BALB/c с фетальным вальпроат-синдромом (ФВС), которым с 7-х по 14-е сут постнатального развития перорально ежедневно вводили афобазол (10 мг/кг) или 0,9% раствор хлорида натрия. Контрольная группа получала 0,9% раствор хлорида натрия в эквивалентном объеме (0,1 мл на 10 г массы). Состояние мышат изучали с 6-х по 14-е сут постнатального развития, оценивали их физическое развитие, скорость созревания сенсорно-двигательных рефлексов, эмоционально-двигательное поведение и точную координацию движений при помощи батареи «развитийных» тестов. Результаты. Введение самкам мышей на 13-й день беременности вальпроевой кислоты приводило к отставанию созревания у потомства сенсорно-двигательных рефлексов, нарушению эмоционально-двигательного поведения и координации движений в гнездовом периоде. Афобазол, при введении 10 мг/кг перорально ежедневно, начиная с 7-х сут постнатального развития мышам с ФВС, корригировал отмеченные нарушения в тестах, отражающих нарушения развития нервной системы. Заключение. Установлены корригирующие свойства афобазола в отношении нарушений, вызванных пренатальным введением ВПК, что определяет целесообразность дальнейшего изучения афобазола на моделях РАС. Introduction. Exposure of rodents to valproic acid during pregnancy is associated with increased incidence of autism spectrum disorders, and has been extensively used as an appropriate model of autism. Aim. To study early behavioral changes in BALB/c mice prenatally exposed to a single dose of valproic acid sodium salt (400 mg/kg) and a possibility of correcting these changes with afobazole. Methods. The study was performed on BALB/c mice with fetal valproate syndrome (FVS). The mice were daily injected orally afobazole 10 mg/kg or 0.9% sodium chloride from day 7 to day 14 of the postnatal development. The control group was injected with an equivalent volume (0.1 ml per 10 g body weight) of 0.9% sodium chloride. The condition of mice was studied from day 6 to day 14 of the postnatal development with evaluation of their physical development, maturation rate of sensory-motor reflexes, emotional-motor behavior, and precise coordination using a battery of «developmental» tests. Results. Administration of valproic acid to female mice on the 13th day of pregnancy led to delayed maturation of the offspring’s sensory-motor reflexes, impaired emotional-motor behavior and coordination of movements during the nesting period. Afobazole administered to mice with fetal valproate syndrome from day 7 to day 14 of the postnatal development at a dose of 10 mg/kg (daily, orally), corrected the disorders in the tests used for assessing retardation or disruption of nervous system development.

Enantiomeric N-substituted phthalimides with excitatory amino acids protect zebrafish larvae against PTZ-induced seizures

Epilepsy is a chronic neurological disease with high prevalence and adverse impacts on the quality of life of patients and caregivers. Up to one-third of individuals with epilepsy do not respond to current pharmacotherapy, underscoring the importance of identifying new molecules for epilepsy control. Thalidomide, the first synthetized phthalimide, is a neuroactive molecule with anti-seizure drug properties. The phthalimide group has been studied in some N-phthaloyl amino acids due to its pharmacological properties. Here we examine enantiomers of phthaloyl aspartate (R and S) and phthaloyl glutamate (R and S) for anti-seizure effects using zebrafish as a model. The zebrafish model is rapidly growing in use as a preclinical screening tool for drug discovery in epilepsy. Pentylenetetrazol (PTZ) exposure was used to produce convulsive behavior in 7- and 10-days post-fertilization (dpf) zebrafish larvae; these ages correspond to before and after the blood-brain-barrier (BBB) is fully developed. Larvae were pre-treated for 60 min with: control, valproic acid sodium salt (SVP) 3 mM, or one of two concentrations of N-phthaloyl-R-glutamic acid (R-TGLU; 100, 316 μM) prior to PTZ addition. R-TGLU modified the locomotor phenotype and protected against PTZ in 7 and 10 dpf larvae at 316 μM, suggesting it crossed the BBB. We next tested the per se and anticonvulsant effect of the glutamate and aspartate phthalimides were tested at 237.1 and 316 μM concentration in 10dpf zebrafish. The four tested molecules produced an anticonvulsant effect at 237.1 μM concentration, however the behavioral changes that they induce suggest that they might act by different mechanisms.

The influences of SCN3A gene polymorphism on the efficacy of valproic acid sodium in the treatment of epilepsies

Objective To investigate the influences of SCN3A gene polymorphism(c.905A>G/p.N302S and c. 1441C>T/p.L481L) on the efficacy of valproic acid sodium in the treatment of Zhuangzu epilepsies. Methods Using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)technique and the way of direct sequence, the SCN3A gene c. 905A>G/p.N302S and c. 1441C>T/p.L481L genotypes in peripheral blood were detected in 244 epileptic patients (85 cases in effective group and 139 cases of ineffective group) in the standardized treatment of valproic acid sodium.The blood concentration of valproic acid sodium was detected by LC-MS.Evaluating the correlation between the genotype and alleles of two groups of patients and the efficacy of valproic acid sodium and analyzing the difference of valproic acid sodium's blood concentration between different genotypes.The linkage disequilibrium of c. 905A>G/p.N302S and c. 1441C>T/p.L481L were analyzed by software SHEsis. Results The allele and genotype distribution in c. 905A>G/p.N302S loci between effective group(A, G allele: 50.6%, 49.4%, AA, AG, GG genotype: 27.1%, 47.1%, 25.8%) and ineffective group(A, G allele: 37.4%, 62.6%, AA, AG, GG genotype: 16.6%, 41.7%, 41.7%) had statistically significant difference(χ2=7.501, P=0.006; χ2=7.907, P=0.019). There was no significant difference in allele and genotype distribution of c. 1441C>T/p.L481L loci between effective group(C, T allele: 47.1%, 52.9%, CC, CT, TT genotype: 23.5%, 47.1%, 29.4%) and ineffective group(C, T allele: 38.8%, 61.2%, CC, CT, TT genotype: 18.7%, 40.3%, 41.0%)(χ2=2.920, P=0.088; χ2=3.099, P=0.212). Compared with the AA + AG genotype, the GG genotype at c. 905A>G/p.N302S locus significantly reduced the efficacy of valproic acid sodium (OR=2.051, 95%CI=1.136-3.703). Compared with genotypes AA+ AG, there were no significant differences in blood concentration of genotype GG of c. 905A>G/p.N302S (t=3.256, P=0.137). Compared with genotypes CC+ CT, there were no significant differences in blood concentration of genotype TT of c. 1441C>T/p.L481L(t=4.628, P=0.082). c.905A>G/p.N302S and c. 1441C>T/p.L481L were without linkage disequilibrium. Conclusion These results suggest that the single nucleotide polymorphisms of c. 905A>G/p.N302S in SCN3A genes may play a role in the resistivity of valproic acid sodium in Zhuangzu epilepsies. Key words: Polymorphism of SCN3A genes; Zhuangzu epilepsie; Valproic acid sodium; Efficacy

Histone deacetylase inhibitors differentially regulate c-Myc expression in retinoblastoma cells.

Retinoblastoma (RB) is the most prevalent childhood intraocular cancer type. Previous studies have demonstrated that c-myc (a proto-oncogene) is associated with tumorigenesis. However, at present, the influence of the expression profile and bioactivity of c-Myc on RB occurrence and progression is yet to be characterised. Notably, the present study demonstrated that c-myc is downregulated in the RB cell line WERI-Rb1. However, treatment with the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) was revealed to significantly upregulate the expression of c-Myc mRNA and protein in WERI-Rb1 cells. Moreover, TSA increased the activity of the c-myc promoter in WERI-Rb1 cells, and the expression of c-Myc was also regulated by other HDAC inhibitors, including vorinostat (SAHA), valproic acid sodium salt (VPA) and entinostat. Notably, although c-myc was silenced in the Y79 cell line, the HDAC inhibitor TSA did not induce upregulation of mRNA and protein in Y79 cells. By contrast, certain HDAC inhibitors (TSA, VPA and SAHA) were discovered to significantly decrease the activity of the c-myc promoter in Y79 cells. Furthermore, the current data indicated that exogenous c-myc expression has a mild inhibitory effect on WERI-Rb1 and Y79 cell viability. Therefore, the present study revealed novel insights into the expression mechanism and bioactivity of c-Myc in RB cells.

The Antiviral Effects of Sodium Phenylbutyrate Against BoHV-1 Infection In Vitro

Introduction: The alteration of histone acetylation is a known mechanism to regulate gene expression, and thereby affecting various cellular processes. Histone deacetylases (HDACs) are known to regulate histone acetylation by removal of the acetyl group from lysines. HDAC inhibitor such as Sodium Phenylbutyrate (PB) and Valproic Acid (VPA) have been reported to affect multiple virus infection while whether they affect BoHV-1 infection is unknown. Objective: The aim of the study is to investigate whether PB and VPA effects BoHV-1 infection and the virus induced inflammation related signaling including Erk1/2 and p38MAPK signaling. Methods: To assess the antiviral effects of PB and VPA on BoHV-1 infection, MDBK cells were treated with these inhibitors at different concentrations. Then time addition was performed to pinpoint which stages of virus infection was affected by the chemicals. In order to assess whether PB affect viral gene expression, we detected the viral IE genes such as bICP0, bICP4 and bICP22 using real-time PCR assay. The effects of PB had on the activation of inflammation related signaling including Erk1/2 and p38MAPK in response to the virus infection were also detected. Results: Here, for the first time we reveals that PB but not VPA affects BoHV-1 infection at late stages of infection. It affected the expression of IE genes such as bICP0, bICP4 and bICP22. Interestingly, PB enhanced the activation of both Erk1/2 and p38MAPK signaling stimulated by BoHV-1 infection. Conclusion: HDAC inhibitor PB significantly inhibited BoHV-1 infection partially through the interruption of certain viral IE gene expression. Though PB has been reported to have antiinflammatory effects, we found that it enhanced the activation of inflammation pertinent signaling of both Erk1/2 and p38MAPK stimulated by BoHV-1 infection.

Twenty-eight-day repeated oral doses of sodium valproic acid increases neural stem cells and suppresses differentiation of granule cell lineages in adult hippocampal neurogenesis of postpubertal rats

Developmental exposure to valproic acid (VPA), a model compound for experimental autism, has shown to primarily target GABAergic interneuron subpopulations in hippocampal neurogenesis of rat offspring. The VPA-exposed animals had revealed late effects on granule cell lineages, involving progenitor cell proliferation and synaptic plasticity. To investigate the possibility whether hippocampal neurogenesis in postpubertal rats in a protocol of 28-day repeated exposure is affected in relation with the property of a developmental neurotoxicant by developmental exposure, VPA was orally administered to 5-week-old male rats at 0, 200, 800 and 900 mg/kg body weight/day for 28 days. At 900 mg/kg, GFAP+ cells increased in number, but DCX+ cells decreased in number in the granule cell lineages. Moreover, CHRNB2+ cells and NeuN+ postmitotic neurons decreased in number in the hilus of the dentate gyrus. Transcript level examined at 900 mg/kg in the dentate gyrus was increased with Kit, but decreased with Dpsyl3, Btg2, Pvalb and Chrnb2. These results suggest that VPA increased type-1 stem cells in relation to the activation of SCF-KIT signaling and suppression of BTG2-mediated antiproliferative effect on stem cells. VPA also decreased type-3 progenitor cells and immature granule cells probably in relation with PVALB+ interneuron hypofunction and reduced CHRNB2+ interneuron subpopulation in the hilus, as well as with suppression of BTG2-mediated terminal differentiation of progenitor cells. Thus, the disruption pattern of VPA by postpubertal exposure was different from developmental exposure. However, disruption itself can be detected, suggesting availability of hippocampal neurogenesis in detecting developmental neurotoxicants in a 28-day toxicity study.

Formulation Development and Evaluation of Sodium Valproate and Valproic acid Extended Release Tablets

The objective of the present study was to formulate and develop extended release drug delivery system of antiepileptic drug Sodium valproate (VAS) and Valproic acid (VA) in the ratio of 70: 30 with extended release profile. Preliminary studies with different polymers such as HPMC K15M, PVP K90, Ethyl cellulose N50, Carbopol were performed. The results of in-vitro release data showed that HPMC K15M can sustain the drug release upto 24hr. From these studies HPMC K15M has been selected for further studies. HPMCK15M was used along with HEC 250HX to retard the drug release. VAS and VA extended release tablets were prepared by wet granulation method. The hardness of these extended release tablets was within the range between 5.63±0.42 to 6.92±0.33 kg/cm2. The drug content was within the range, 97.23±0.25 to 102.03±2.45%. The in-vitro VAS and VA release from the tablets was found sustained over 24 hours with Higuchi kinetics of drug release and release pattern followed anomalous (Non-Fickian) diffusion. The Fourier transform Infrared spectroscopy analyses indicated that there was absence of chemical interaction between the drug and the excipients. The dissolution profiles of the developed formulation and the commercial tablet formulation, Encorate, were compared using the similarity factor (f2) and difference factor (f1). The released profile of tablet containing 15% HPMC K15M and 15% HEC 250HX by weight was similar to that of Encorate® providing the values of similarity factor (f2) and difference factor (f1) of 88.20 and 3.22 respectively.

The Epigenetic Regulation of Blinatumomab Gene Expression: Tumor Cell-dependent T cell Response against Lymphoma Cells and Cytotoxic Activity.

Conventional treatment for cancer such as surgical resection and chemotherapy can cause damage in cases with advanced cancers. Moreover, the identification of tumor-specific targets has great importance in T-cell therapies. For decades, T cell activity has been stimulated to improve anti-tumor activity. Bispecific antibodies have attracted strong interest from pharmaceutical companies, for their diagnostic and therapeutic use. Blinatumomab is a first-in-class bispecific T engager antibody for the treatment of relapsed or refractory precursor B- cell acute lymphoblastic leukemia. But, it can benefit several cases with CD19+ malignancies in the future. PhiC31 integrase-based vectors could selectively integrate therapeutic transgenes into pseudo-attP sites in CHO genome. In this study, production of Blinatumomab in CHO cells using this type of vectors was investigated. We evaluated the effects of histone deacetylases (HDACs) inhibitors such as sodium butyrate and valproic acid, on specific productivity and cell viability of antibody expressing cells. Although sodium butyrate increased specific productivity about 1.7-fold and valproic acid about 1.4-fold, valproic acid was found more efficient because of its less cytotoxic effect on cell growth. We examined the efficacy of expressed Blinatumomab at various effector to target (E/T) ratios. A dose-response analyses of calcein-acetoxymethyl release assay illustrated that the effective dose of expressed mAb required for antibody mediated cytotoxicity was 100 ng/ml and the expressed mAb was more effective at E/T ratios of 10:1 and 5:1. Results of this study indicated that the expressed blinatumomab can be useful for enhancing the cytotoxicity of CD3+ T-cells against CD19 + target cells in vitro.

Photocatalytic treatment of valproic acid sodium salt with TiO2 in different experimental devices: An economic and energetic comparison

Valproic acid sodium salt (VA) removal by heterogeneous photocatalysis was investigated in four different devices. An economic and energetic comparison was performed with three reactors based on artificial irradiation (BLB, black light blue lamps, λ=365nm; UVC, λ=254nm; SB, Xe lamp, solar simulator) and a solar pilot plant based on compound parabolic collectors (CPC). For the highest TiO2 concentration used (0.4g/L), the best results in VA degradation and TOC removal were obtained for CPC (91% and 50%, respectively). Costs related to electricity consumption for operation and sampling were considered for this comparison. The lower cost per treated volume (0.48€/L) and per mg VA treated (0.012€/mg) also correspond to CPC device. However, BLB gives the best result in cost per ppm reduction (0.027€/ppm), which shows the importance to have a clear specification of the considered volume and of the total volume treated. Concerning the energetic questions, in the case of laboratory scale, the energy spent in analysis can represent up to 90% of total energy. The efficiency in ppm reduction is higher for BLB (4.77ppm/kWh) while the highest efficiency in mg of pollutant reduction corresponds to CPC (11.12mg/kWh). Summarizing, the use of more adequate radiation and geometry make BLB and CPC good alternatives for the efficient treatment of VA.