Chrysin ameliorates seizures, seizure-induced oxidative stress and cognitive impairment in experimental models of epilepsy in rats

Chrysin ameliorates seizures, seizure-induced oxidative stress and cognitive impairment in experimental models of epilepsy in rats | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Chrysin ameliorates seizures, seizure-induced oxidative stress and cognitive impairment in experimental models of epilepsy in rats Dr Sunil Kumar, Professor K.H. Reeta, Dr Pankaj Prabhakar, Dr Santenna Chenchula This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8407071/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Purpose: Chrysin (CH), a naturally occurring flavonoid found in various traditional medicinal plants and bee products, has been reported to possess antioxidant and neuroprotective properties. These properties may offer therapeutic potential in epilepsy, a chronic neurological disorder frequently accompanied by cognitive impairment and oxidative stress. To evaluate the anticonvulsant, antioxidative, and cognitive-protective effects of chrysin in experimental models of epilepsy. Materials and methods: The effects of CH were assessed using three established rodent models: pentylenetetrazol (PTZ)-induced seizures, maximal electroshock (MES)-induced seizures, and PTZ-induced kindling. Seizure severity, protection percentage, and progression of kindling were recorded. Cognitive performance was evaluated across all models, and biochemical assays were conducted to measure oxidative stress markers and acetylcholinesterase (AChE) activity. Results: CH (120 mg/kg) demonstrated robust anticonvulsant activity, offering 83.3% protection against generalized tonic–clonic seizures in the PTZ model and complete (100%) protection in the MES model. In PTZ-induced kindling, CH significantly attenuated epileptogenesis (mean seizure score = 1.66). All three models exhibited marked cognitive deficits, which were significantly improved by CH treatment. CH also effectively restored oxidative stress parameters and reversed the seizure-induced reduction in AChE activity. Conclusions: Chrysin exhibits potent anticonvulsant and neuroprotective effects in multiple seizure models, mitigating seizure severity, oxidative stress, and associated cognitive deficits. These findings support the ethnopharmacological relevance of chrysin-containing natural products and highlight CH as a promising phytochemical candidate for managing epilepsy and its neurocognitive complications. Chrysin Epilepsy Pentylenetetrazol Maximal electroshock seizure Oxidative stress Cognitive impairment Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Introduction Epilepsy is one of the most prevalent and leading neurological disorders, with an incidence of about 1% of the population [1]. Moreover, it affects about 50 million people worldwide, and almost 2.4 million people are diagnosed with epilepsy every year [2]. The prevalence of epilepsy is higher in developing countries than in developed countries [3]. Many biological events contribute to the development and pathogenesis of epilepsy, including the inhibition of gamma-aminobutyric acid (GABA), a naturally occurring amino acid that functions as a neurotransmitter in the brain, as well as imbalanced inhibitory/excitatory transmissions and altered regulatory modulator systems [4]. Cognitive impairment with abnormal behaviors in patients with epilepsy is observed, and a range of factors responsible for this, like anticonvulsant drugs, seizure type/frequency, structural lesions, and psychosocial stress [5]. In addition, oxidative stress (OS), which is associated with alterations in reactive oxygen species (ROS), reactive nitrogen species (RNS), and nitric oxide (NO) signaling pathways, whereby bioavailability of NO is decreased, and ROS and RNS production are increased. Further, oxidative damage is very common in the brain due to the abundance of mitochondria, high oxygen demand, poor repair capacity and high concentrations of polyunsaturated fatty acids, which play an important role in the neurological disorders, including epilepsy [6]. Moreover, there is involvement of the OS in the pathogenesis and progression of seizures, which adds to the cognitive disorders in patients with epilepsy [7]. It has been demonstrated that there is change in cholinergic transmission in cognition impairment caused by seizure and assessment of cholinergic status in rat brain after seizures which can be done by evaluation of change in acetylcholinesterase activity too [8]. There is a significant number of antiepileptic drugs (AEDs) in the clinics for epileptic seizures, but these therapies target only symptoms of the diseases. They failed to adequately prevent seizure development or permanently halt the occurrence of seizures [9]. Moreover, AEDs is responsible for various side effects such as dizziness, tremor (shakiness), drowsiness, unsteadiness, blurred or double vision, bruising, and bleeding [10]. In additon, more than 30% patients with epilepsy refractory to AEDs, which leads to a significant increase in the morbidity, mortality and impairment in quality of life in refractory patients of epilepsy [11]. There is an urgent need for newer and safer drugs to combat multi-factorial neurochemical abnormalities related to epileptic seizures. Flavonoids, a subclass of polyphenols, are synthesised by plants as a secondary metabolite and are widely present in fruits, vegetables, and certain beverages. It has been demonstrated that flavonoids exhibit numerous beneficial effects in patients with epilepsy without causing any adverse side effects [10]. It has been demonstrated that the beneficial effects of flavonoids in neurological diseases may be associated with the modulation of GABA receptors, various voltage-gated ion channels, the regulation of antioxidant pathways, anti-inflammatory mediators, and the modulation of mitochondrial dysfunction and cytokine activity [10]. Chrysin (CH) (5,7-dihydroxy-2-phenyl-4H-chromen-4-one), a flavonoid, is a natural and most important bioactive constituent of honey, many fruits (like passion fruit (Passiflora sp.), kiwi, strawberries, etc.), vegetables and mushrooms [12, 13]. CH possesses an array of pharmacological properties such as antioxidative, neuroinflammatory, anti-depressant, anti-amyloidogenic, anticancer, and neuroprotective effects [12, 13]. In view of these aforementioned facts, we designed this study to evaluate the effect of chrysin in pentylenetetrazol (PTZ) and maximal electroshock seizure (MES)-induced seizure, PTZ-induced kindling, seizures-induced oxidative stress and cognitive impairment in male Wistar rats. Materials and methods Animals The study protocol was approved by the Institutional Animal Ethics Committee of the All-India Institute of Medical Sciences (AIIMS), New Delhi, India (843/IAEC/15). Male Wistar rats, weighing 150–225 g, were obtained from the Central Animal Facility at AIIMS, New Delhi, India. They were maintained under standard laboratory conditions at a temperature of 25 ± 2°C with a natural light-dark cycle, and a standard dry rat pellet diet and tap water were provided ad libitum . The rats were acclimated to the laboratory environment for a week, and the animals were randomly assigned to the experimental groups, consisting of six rats per group, after 7 days of acclimation. Drugs and Chemicals Chrysin, pentylenetetrazol, sodium valproate and all chemicals used for the experiment were of analytical grade and were purchased from Sigma-Aldrich Co., USA. Pentylenetetrazol (PTZ)-induced seizures PTZ solution (60 mg/kg, once) was freshly prepared in normal saline and administered intraperitoneally (i.p.), 30 minutes and 60 minutes after the administration of sodium valproate (300 mg/kg; i.p.), chrysin (CH) (in doses of 30 mg/kg, 60 mg/kg and 120 mg/kg; once orally) respectively. The dose of PTZ (60 mg/kg, once) was standardised in our laboratory as the 100% convulsant dose with minimal mortality in rats [ 14 ]. We recorded the latency to myoclonic jerks and the incidence of generalised tonic–clonic seizures (GTCS) with loss of righting reflex. Rats were monitored for 30 minutes after administration of PTZ. We also evaluated the chronic effect of CH (in the doses of 30 mg/kg, 60 mg/kg and 120 mg/kg; once daily; orally) in a chronic epileptic seizure (kindling) rat model, which was induced by administration of sub-convulsive doses of PTZ (30 mg/kg; i.p.) on alternate day (48 ± 1 h). PTZ was administered up to day 52 or until seizure stage 5 on two consecutive trials were achieved, whichever was earlier. Sodium valproate (300 mg/kg; i.p.) was also used as a standard. The rats were observed for 30 min after each PTZ administration for convulsive behaviour. Seizure activity was evaluated according to the method reported by Racine [ 15 ]. The latencies to myoclonic jerks and GTCS were recorded and were transformed into seizure score (S) [ 16 ], which was calculated using the following formula: S = 1- (control latency/drug seizure latency) Rats were considered kindled if they exhibited stage 5 of seizures on two consecutive trials. For control rats, S = 0 and S = 1 for rats who did not develop seizures. Rats were also observed for 24 h mortality, if any. Maximal electric shock (MES)-induced seizures Rats were randomly divided into six groups. The first group served as vehicle control, the second group was the MES-induced seizure group, and the third group was treated with standard drug, i.e. sodium valproate (300 mg/kg; i.p.). The fourth, fifth, and sixth groups were administered CH 30 mg/kg, 60 mg/kg and 120 mg/kg, respectively. Electroconvulsions were produced in rats by a suprathreshold fixed-current sinus wave stimulus (70 mA, 0.2-second duration) delivered via ear-clip electrodes (Ugo Basile, Italy), 30 minutes and 60 minutes after administration of sodium valproate and CH, respectively. Rats were observed for the occurrence of tonic hind limb extension (THLE), i.e., the hind limbs of rats outstretched 180 o to the plane of the body axis [ 5 ]. Cognitive function assessment The behavioural parameters were performed before and 24 h after induction of seizures caused by PTZ or MES, and this was followed by CH administration. Morris water maze (MWM), elevated plus maze (EPM) and passive avoidance test (PAT) were conducted to assess any behavioural changes of the rats. Sixty minutes after CH administration, seizures were induced with either PTZ or MES. During the behavioural experiments, only one animal was tested at a time. Moreover, behavioural parameters were evaluated at baseline and after completion of the kindling procedure in the kindling experiment. Elevated plus maze test The elevated plus maze (EPM) test was performed to assess acquisition and retention memory processes as described earlier [ 17 ]. The maze was placed 50 cm above the floor and consists of two closed arms and two open arms forming a cross, with a quadrangular centre. On day 1, the initial transfer latency was measured as follows: the rats were placed individually at the end of one open arm, facing away from the central platform, and the time it took to move from the open arm to either of the enclosed arms was recorded. Transfer latency was the time it took for the rats to move from the open arm to the enclosed arm, when all four of their legs crossed over to the enclosed arm. The rat was gently pushed onto its back into the enclosed arm, and a transfer latency of 60 seconds was assigned if the rat did not enter the enclosed arm within 60 seconds. After measuring the transfer latency, the rat was allowed to move freely in the plus maze, regardless of whether the arms were open or closed, for 10 seconds. The rat was then gently removed from the plus maze and returned to its home cage. Twenty-four hours later, the retention transfer latency test was performed in the same manner as in the acquisition trial. If the rat did not enter the enclosed arm within 60 seconds on the second trial, the transfer latency was assigned 60 seconds. Morris water maze Morris water maze was used to test the spatial memory of the rats. The testing procedure was used as described earlier [ 17 ]. Briefly, the water tank was divided into four equal quadrants (Q1, Q2, Q3 & Q4) and the platform was kept in the fourth quadrant. Rats were given four trials during four daily acquisition sessions. Every trial began by placing a rat in a water tank, facing the wall of the tank. Each of the four starting points was used once in a series of trials. The location of the platform was fixed for each trial. Further, the trial was terminated automatically as soon as the rat reached the platform or when 120 s had elapsed. The rat was allowed to continue on the platform for 5 seconds. Then it was removed from the platform, and a new trial was initiated. Rats that did not locate the platform within 120 seconds were gently placed on the platform and allowed to stay there for 5 seconds. After each trial, the rats were gently dried with a towel and returned to their home cage. On the fifth day, a spatial probe trial (60 seconds) was performed to assess the rat's spatial memory. On the 5th day, the same protocol was followed as described above; however, the platform had been removed from the tank. The path of each rat was analysed using the Any-maze video tracking system (Caterpillar Instrumentation Pvt. Ltd.). The latency to reach the platform and the time spent in the target quadrant (in which the platform was kept) were recorded. . One-trial passive avoidance task Memory retention deficit was evaluated by a step-through passive avoidance task [Ugo Basile, Italy (Model 7552)] as previously described [ 17 ]. Briefly, on the acquisition trial, each rat was placed in a lighted chamber. After 60 seconds of habituation, a guillotine door separating the lighted and dark chambers was opened, and the initial latency (IL) to enter the dark chamber was recorded. Rats exhibiting an initial latency time of more than 60 s were excluded from further experiments. Immediately after the rat entered the dark chamber, the guillotine door was closed, and an electric foot shock (75 V, 0.2 mA, 50 Hz) was delivered to the floor grids for 3 s. The rat was then removed from the dark chamber 5 seconds later and returned to its home cage. After 24 hours, retention latency (RL) time was measured in the same manner as during the acquisition trial, but foot shock was not delivered, and the latency time was recorded up to a maximum of 600 seconds. Biochemical estimation At the end of the experiments, all rats were sacrificed, and samples of brain tissue were collected, cleaned with ice-cold saline and stored at − 80°C until further analysis. Tissue preparation Brain tissue samples were thawed, and the cortex and hippocampus from each cerebral hemisphere were separated for biochemical estimations. From separated parts (cortex and hippocampus), 10% w/v homogenates were prepared with ice-cold 0.1 M phosphate buffer (pH 7.4). Aliquots were prepared to determine lipid peroxidation, reduced glutathione, superoxide dismutase (SOD), acetylcholinesterase (AChE), nitric oxide, and protein estimation. The other half was preserved. Measurement of lipid peroxidation TBARS (Thiobarbituric acid reactive substances), a measure of lipid peroxidation, was estimated as described earlier [ 18 ]. Briefly, acetic acid 375 µl (20%; pH 3.5), 375 µl thiobarbituric acid (0.8%), 50 µl sodium dodecyl sulfate (8.1%) and 20 µl water were added to 30 µl of processed brain tissue samples (cortex and hippocampus). The mixture was heated at 95°C for 60 minutes. The mixture was then cooled, and 1,000 µL of a 15:1 (v/v) n-butanol: pyridine solution was added. The mixture was then shaken vigorously. After centrifugation at 4000 rpm for 10 min, the organic layer was withdrawn, and absorbance was measured at 532 nm using a spectrophotometer. Measurement of reduced glutathione (GSH) Reduced glutathione was estimated as described by Ellman [ 19 ]. Briefly, the homogenate (cortex and hippocampus) was centrifuged with 5% trichloroacetic acid to precipitate the proteins. To 50 µl of this supernatant and 200 µl of 0.3 M phosphate buffer (pH 8.4), 25 µl of 5′5 dithiobis (2-nitrobenzoic acid) (DTNB) was added. The mixture was then vortexed, and the absorbance was read at 412 nm within 15 min. Estimation of nitrite The accumulation of nitrite in the brain tissue (cortex and hippocampus) supernatant, an indicator of the production of nitric oxide (NO), which was determined as described by Green and his coworkers [ 20 ], with Greiss reagent [0.1% N-(1-naphthyl) ethylenediamine dihydrochloride, 1% sulfanilamide, and 5% phosphoric acid]. Equal volumes of the supernatant (50 µL) and the Griess reagent (50 µL) were mixed, and the mixture was incubated at room temperature for 10 minutes. The absorbance was determined at 540 nm on an ELISA reader. The concentration of nitrite in the supernatant was determined from a sodium nitrite standard curve and was expressed as micromole per gram. Superoxide dismutase (SOD) Superoxide dismutase was estimated as described by Marklund and Marklund [ 21 ]. Briefly, homogenized tissues with 10 times (w/v) 0.1 M sodium phosphate buffer (7.4). The reagent Tris-HCl-EDTA buffer (900 µl, pH 8.2) was added to 50 µl of the processed sample. Finally, at the time of reading, pyragallol (50 µL, 4 mM) was added, and optical observations were taken at 420 nm at different time intervals. The concentration of SOD was expressed as Units per Milligram of protein. Estimation of Cholinergic Status The cholinergic markers, acetylcholinesterase, were estimated in the cortex and hippocampus of the brain tissue as described earlier by Ellman [ 22 ]. Briefly, the rats' brains were removed over ice. The tissue was then homogenised in 0.1 M phosphate buffer and centrifuged at 10000 rpm for 30 min. at 4°C. 25 µl of this supernatant was incubated for 10 min with 650 µl buffer and 25 µl of DTNB (10 µM). Then, 25 µl of the freshly prepared acetylthiocholine iodide (75 mM) was used as substrate for the estimation of AChE. The absorbance was read at 412 nm for 3 min at 30, 60, 90, 120, 150 and 180 seconds. Statistical analysis Results are represented as mean ± standard error of the mean (SEM). One-way analysis of variance (ANOVA) followed by the Bonferroni multiple comparison test (SPSS-23 software) was done. A value of P < 0.05 was considered statistically significant. Results Effect of chrysin on Pentylenetetrazol (PTZ) induced seizures PTZ administration caused myoclonic jerks latency (54.6 second ± 6.1 second) (Table 1 ) and generalized tonic-colonic seizure latency (GTCS; 71.2 second ± 8.30 second) which was prevented by CH in dose dependent manner [myoclonic jerks (CH 30 mg/kg; 92.2 second ± 5.42 second), (CH 60 mg/kg; 127.7 second ± 6.7 second; P < 0.01) and (CH 120 mg/kg; 136 second ± 7.9 second; P < 0.001); GTCS (CH 30 mg/kg; 102 second ± 5.0 second), (CH 60 mg/kg; 150 second ± 9.4 second; P < 0.01), (CH 120 mg/kg; 166 second ± 8.2 second; P < 0.001] (Table 1 ). Moreover, the % age protection against PTZ-induce seizures was observed with CH 30 mg/kg, 60mg/kg and 120 mg/kg in dose dependent manner 66.7%, 83.3% and 100% respectively as compared to PTZ (0% protection). In addition, the %age protection against MES-induce seizures was also observed with CH 30 mg/kg, 60 mg/kg and 120 mg/kg in dose dependent manner 16.7%, 50% and 83.3% respectively as compared to MES (0% protection). Further, there was 100% protection against PTZ and MES in sodium valproate treated rats (Table 1 ). Table 1 Effect of the Chrysin (CH) on myoclonic jerk latency and occurrence of generalized tonic clonic seizures and tonic hindlimb extension in the pentylenetetrazol (PTZ) and maximal electric shock (MES)-induced seizure in rats (n = 6 per group). Values are presented as mean ± SEM. ***P < 0.001 Group Myoclonic jerk latency (s) (mean ± SEM) Latency to onset of GTCS (s) (mean ± SEM) % Protection against Generalized tonic–clonic seizures Tonic hind limb extension PTZ (60 mg/kg) 54.6 ± 6.1 71.2 ± 8.3 0 --- Maximal electric shock (70 mA) --- --- --- 0 Sodium valproate (300 mg/kg) --- --- 100 100 CH30 92.2 ± 5.4 *** 102 ± 5.0 66.7 16.7 CH60 127.7 ± 6.7 *** 150 ± 9.4 ** 83.3 50 CH120 135.9 ± 7.9 *** 165.8 ± 8.2 *** 100 83.3 Chronic administration of PTZ resulted in a gradual increase in seizure score over the course of the experiment, reaching a score of 4.33 ± 0.2. CH administration prevented the seizure severity in dose dose-dependent manner as compared to PTZ [(CH 120 mg/kg, 1.66 ± 0.2 vs 4.33 ± 0.2; P < 0.001), (CH 60 mg/kg, 2.6 ± 0.4 vs 4.33 ± 0.2; P < 0.01), (CH 30 mg/kg, 3.5 ± 0.5 vs 4.33 ± 0.2)]. Sodium valproate significantly protected the rats against PTZ-induced kindling (1 vs 4.33 ± 0.2, P < 0.001) (Fig. 1 ). Therefore, the data from this study demonstrate that CH administration decreases the level of epileptic activity. Effect of CH on Maximal electroshock seizures (MES) induced seizures There was 100%, 50%, and 16.7% protection with CH 120 mg/kg, CH 60 mg/kg, and CH 30 mg/kg, respectively, against MES-induced THLE. None of the animals in the sodium valproate (VAL; 300 mg/kg)-treated group exhibited THLE ( Table 1 ) . Cognitive function Elevated plus maze (EPM) There was no significant difference in the initial transfer latency among the groups, in all three models of epilepsy (PTZ, MES and PTZ-induced kindling). PTZ administration caused a significant increase in retention transfer latency (p < 0.001) compared to the normal control (NC). Pretreatment with chrysin (30, 60 and 120 mg/kg) for 28 days significantly prevented the increase in retention transfer latency (p < 0.001) in a dose-dependent manner as compared to PTZ (60 mg/kg) (Fig. 2 A), which was comparable to sodium valproate. In addition, MES caused a significant increase in the retention transfer latency (p < 0.001) compared to the normal control (NC). Pretreatment with chrysin (30, 60, and 120 mg/kg) for 28 days significantly (p < 0.001) decreased the retention transfer latency in a dose-dependent manner compared to MES ( Fig. 2 B ) . Moreover, in the chronic model, PTZ-induced kindling resulted in a significant increase in retention transfer latency (p < 0.001) compared to the normal control (NC). Administration of chrysin (CH 30 mg/kg, 60 mg/kg, and 120 mg/kg; once daily) for 52 days significantly prevented the increase in retention transfer latency compared to PTZ (30 mg/kg), which was comparable to sodium valproate ( Fig. 2 C ). CH had no significant effect on retention transfer latency in all three epilepsy models compared to the control. Thus, CH improved memory retention, as indicated by significant decreases in retention transfer latency compared to seizure-induced rats. Morris Water maze (MWM) test There was a significant decrease in escape latency from days 1 to 4 in all groups, but no significant differences in escape latency were observed between groups on any of the four days in all three epilepsy models. There was a significant increase in escape latency (P < 0.001) and a decrease (P < 0.001) in time spent in the target quadrant in all three models of epilepsy as compared to the normal control. In the PTZ (60 mg/kg) model, pretreatment with CH significantly (P < 0.001) decreased the escape latency and increased the time spent in the target quadrant in a dose-dependent manner as compared to PTZ (Fig. 3 A and 4 A). The effect of CH120 was comparable with that of sodium valproate ( Fig. 5 A-B ) . However, CH per se did not cause any significant change in either parameter in the MWM test compared to the control. In the MES model, chrysin pretreatment significantly decreased escape latency and increased time spent in the target quadrant in a dose-dependent manner as compared to the MES group. However, CH per se did not cause any significant change in either parameter in the MWM test compared to the control ( Fig. 3 B and 4 B ). In PTZ-induced kindling, CH (CH-60 mg/kg and 120 mg/kg, p.o, once daily) administration for 52 days significantly prevented the increase in escape latency, and CH (120 mg/kg) significantly prevented the decrease in the time spent in the target quadrant caused by PTZ. However, CH per se did not cause any significant change in either parameter in the MWM test compared to the control ( Fig. 3 C and 4 C ). Passive avoidance test There was no significant difference in the initial latency among the different groups, in all three models of epilepsy (PTZ, MES and PTZ-induced kindling). PTZ (60 mg/kg) administration resulted in a significant decrease in retention latency compared to the normal control (P < 0.001). Pretreatment with chrysin (30, 60 and 120 mg/kg) for 28 days significantly increased the retention latency in a dose-dependent manner as compared to PTZ (Fig. 5 A), which was comparable to valproate. CH per se did not cause any significant change in retention latency as compared to control. Subsequently, MES caused a significant decrease in the retention latency as compared to the normal control. Moreover, pretreatment with CH (30, 60, and 120 mg/kg) significantly prevented the decrease in retention latency in a dose-dependent manner caused by MES (Fig. 5 B). However, CH per se did not cause any significant change in retention latency compared to the control. In the chronic model of PTZ-induced kindling, PTZ administration (30 mg/kg) caused a significant decrease in retention latency compared to the normal control. CH (30 mg/kg, 60 mg/kg, and 120 mg/kg, p.o., once daily) administration caused a significant increase in retention latency compared to PTZ. However, chrysin per se did not cause any significant change in retention latency as compared to control (Fig. 5 C ) . Biochemical evaluation Thiobarbituric acid reactive substances (TBARS) levels A significant increase in TBARS levels was observed in brain tissues (cortex and hippocampus) in acute [PTZ (60 mg/kg) and MES] and chronic (PTZ-induced kindling) models of epilepsy, as compared to the normal control group. However, pretreatment with CH significantly attenuated the increased TBARS levels in a dose-dependent manner compared to PTZ, and the effect of CH was comparable to that of valproate (Fig. 6 A). In the MES model, pretreatment with CH significantly prevented the dose-dependent increase in TBARS levels in the cortex and hippocampus caused by MES. The effect of CH on TBARS levels was comparable to that of valproate (Fig. 6 B). In the case of the chronic model (PTZ-induced kindling), administration of CH at a dose of 120 mg/kg significantly prevented the increase in TBARS levels in both the cortex and hippocampus caused by PTZ, which was comparable to that of valproate. Moreover, CH per se did not cause any significant change in the levels of TBARS in all three epilepsy models compared to the control (Fig. 6 C). Reduced glutathione levels There was a significant decrease in the levels of reduced glutathione in the cortex and hippocampus in acute [PTZ (60 mg/kg) and MES] and chronic (PTZ-induced kindling) models of epilepsy as compared to the normal control group. However, pretreatment with three doses of CH (30 mg/kg, 60 mg/kg, and 120 mg/kg) significantly prevented the decrease in reduced glutathione levels in a dose-dependent manner in both the cortex and hippocampus, which was comparable to that of valproate (Fig. 7 A). In the case of the MES model, CH pretreatment also significantly restored the levels of reduced glutathione in both the cortex and hippocampus in a dose-dependent manner, which were comparable to those of valproate (Fig. 7 B). In the case of PTZ-induced kindling, administration of CH at 60 mg/kg and 120 mg/kg significantly prevented the decrease in reduced glutathione levels in the cortex and hippocampus in a dose-dependent manner, as compared to PTZ, which was also comparable to the effect of valproate (Fig. 7 C). In addition, CH per se had no significant effect on reduced glutathione levels in both the cortex and hippocampus in all three epilepsy models, compared to the control. Superoxide dismutase (SOD) levels It was observed that the levels of SOD in the cortex and hippocampus significantly decreased in both the acute and chronic models of epilepsy as compared to the normal control. However, pretreatment with CH significantly restored the levels of SOD in the cortex and hippocampus in a dose-dependent manner, as compared to PTZ (60 mg/kg), which were comparable to those of valproate (Fig. 8 A). In the case of the MES model, pretreatment of CH significantly prevented the decrease in the levels of SOD in the hippocampus, only in a dose-dependent manner, which was comparable with valproate. CH did not cause any significant change in the levels of SOD in the cortex as compared to MES (Fig. 8 B). In the case of PTZ kindling, administration of CH significantly attenuated the decreased levels of SOD in the cortex and hippocampus in a dose-dependent manner, compared to PTZ, and this effect was also comparable to that of valproate. Moreover, CH per se did not cause any significant change in three different models of epilepsy as compared to control (Fig. 8 C). Nitric oxide (NO) levels There was significant increase in the level of nitric oxide (NO) in all three model of epilepsy in cortex and hippocampus. Pretreatment of CH significantly prevented the increase in the levels of NO caused by PTZ (60 mg/kg) in a dose dependent manner in both cortex and hippocampus (Table 2 ). Table 2 Effect of CH on cortex and hippocampus nitric oxide (NO) levels in the A) PTZ- seizure model B) MES- seizure model C) PTZ-induced kindling in rats; (n = 6 per group). Values are presented as mean ± SEM. *P < 0.05, ***P < 0.001; a- as compared with the control; b- as compared with the PTZ group and MES group. Groups NO levels (µg/g of wet tissue) in cortex in acute PTZ NO levels (µg/g of wet tissue) in hippocampus in acute PTZ NO levels (µg/g of wet tissue) in cortex in MES model NO levels (µg/g of wet tissue) in hippocampus in MES model NO levels (µg/g of wet tissue) in cortex in chronic PTZ model NO levels (µg/g of wet tissue) in hippocampus in chronic PTZ model Control 48.0 ± 2.6 52.53 ± 2.2 47.63 ± 2.5 52.0 ± 2.5 48.35 ± 4.0 54.87 ± 3.5 PTZ (60 mg/kg) 308.96 ± 10.7 ***a 333.43 ± 11.0 ***a --- --- --- --- MES (70 mA) --- --- 166.80 ± 6.3 ***a 180.94 ± 15.5 ***a --- --- PTZ (Kindling) --- --- --- --- 168.78 ± 8.1 ***a 143.42 ± 6.92 ***a Sodium valproate (300 mg/kg) 116.53 ± 5.6 ***b 80.43 ± 4.9 ***b 44.55 ± 12.1 ***b 61.02 ± 2.8 ***b 69.81 ± 5.7 ***b 68.54 ± 7.0 ***b CH30 144.36 ± 11.0 ***b 96.26 ± 8.8 ***b 97.25 ± 6.4 ***b 107.9 ± 7.8 ***b 147.35 ± 5.5 125.12 ± 6.5 CH60 59.6 ± 3.3 ***b 71.2 ± 4.2 ***b 44.01 ± 3.3 ***b 64.30 ± 6.0 ***b 126.05 ± 8.4 *b 105.18 ± 11.3 *b CH120 32.80 ± 4.8 ***b 39.63 ± 4.8 ***b 29.25 ± 3.9 ***b 59.32 ± 5.8 ***b 65.74 ± 6.3 ***b 77.96 ± 7.6 ***b CH per se 44.73 ± 4.9 58.57 ± 6.2 48.25 ± 4.2 54.73 ± 2.1 50.29 ± 6.2 49.23 ± 2.1 In case of MES model, CH pretreatment significantly prevented the increase in the levels of NO in cortex and hippocampus in dose dependent manner caused by MES (Table 2 ). In case of PTZ-induced kindling, administration of CH (120 mg/kg) significantly restored the levels of NO in cortex and hippocampus in dose dependent manner as compared to PTZ. CH at 30 mg/kg did not significantly decrease NO levels in cortex and hippocampus as compared to PTZ group (Table 2 ). The effect of CH was comparable to that of valproate in all three epilepsy models. Moreover, chrysin per se did not cause any significant change as compared to the control in all three different models of epilepsy. Acetylcholinesterase (AChE) levels PTZ (60 mg/kg), MES and PTZ-induced kindling caused significant decrease in the activity of AChE in both cortex and hippocampus as compared to normal control. Pretreatment with CH significantly increased the AChE activity in cortex and hippocampus in dose dependent manner as compared to PTZ. However, CH at the dose of 30 mg/kg and 60 mg/kg did not significantly prevent the decrease in AChE activity caused by PTZ (Table 3 ). Table 3 Effect of CH on cortex and hippocampus acetylcholinesterase (AChE) levels in the A) PTZ- seizure model B) MES- seizure model C) PTZ-induced kindling in rats; (n = 6 per group). Values are presented as mean ± SEM. ***P < 0.001; a- as compared with the control; b- as compared with the PTZ group and MES group. Groups AChE levels (Units) in cortex in acute PTZ AChE levels (Units) in hippocampus in in acute PTZ AChE levels (Units) in cortex in in MES model AChE levels (Units) in hippocampus in in MES model AChE levels (Units) in cortex in chronic PTZ model AChE levels (Units) in hippocampus in chronic PTZ model Control 2.81 ± 0.1 2.11 ± 0.2 2.84 ± 0.1 2.30 ± 0.1 2.62 ± 0.09 2.69 ± 0.1 PTZ (60 mg/kg) 0.56 ± 0.03 ***a 0.61 ± 0.1 ***a --- --- --- --- MES (70 mA) --- --- 0.45 ± 0.06 ***a 0.55 ± 0.1 ***a --- --- PTZ (Kindling) --- --- --- --- 0.70 ± 0.1 ***a 0.64 ± 0.2 ***a Sodium valproate (300 mg/kg) 1.41 ± 0.1 **b 2.71 ± 0.1 ***b 1.75 ± 0.1 ***b 2.03 ± 0.1 ***b 2.24 ± 0.2 ***b 2.07 ± 0.1 ***b CH30 0.60 ± 0.03 0.67 ± 0.08 0.99 ± 0.04 1.03 ± 0.1 1.04 ± 0.07 0.71 ± 0.04 CH60 1.00 ± 0.07 1.04 ± 0.1 1.12 ± 0.1 *b 1.44 ± 0.1 ***b 1.69 ± 0.06 ***b 1.13 ± 0.08 ***b CH120 1.40 ± 0.04 **b 2.16 ± 0.2 ***b 2.21 ± 0.2 ***b 1.86 ± 0.1 ***b 1.72 ± 0.1 ***b 1.58 ± 0.07 ***b CH per se 2.56 ± 0.3 1.99 ± 0.3 2.61 ± 0.1 2.29 ± 0.7 2.34 ± 0.2 2.41 ± 0.1 In case of MES model, CH pretreatment significantly reversed the activity of AChE in both cortex and hippocampus in dose dependent manner as compared to MES challenges rats. CH at 30 mg/kg did not significantly reverse the decrease in AChE activity caused by MES (Table 3 ). In case of PTZ-induced kindling model, administration of CH significantly prevented the decrease in AChE levels in both cortex and hippocampus as compared to PTZ group. CH at 30 mg/kg did not significantly prevent the decrease in AChE activity caused by PTZ (Table 3 ). The effect of CH was comparable with valproate in all three different model of epilepsy. CH per se had no significant effect on AChE activity as compared to control in all three different model of epilepsy. Discussion Epilepsy is one of the common neurological disorders which still requires antiepileptic drugs (AEDs) with fewer side effects. A large number of antiepileptic drugs (AEDs) are available for the treatment of epilepsy, but treatment is still not satisfactory due to their side effects [ 23 ]. In addition, it has also been reported that epilepsy is responsible for cognitive impairment. Moreover, AEDs have also been shown to induce cognitive impairment in patients with epilepsy [ 24 , 25 ]. CH is a natural flavonoid which occurs naturally in many plants, honey, and propolis. It possesses many biological activities and pharmacological effects, such as antioxidant, anti-inflammatory, anticancer, and antiviral activities [ 12 , 26 ]. In addition, CH exhibits neuroprotective properties by inhibiting apoptosis [ 13 , 26 ]. The present study was conducted to evaluate the effect of CH in acute [induced chemically (PTZ; 60 mg/kg) and electrically (MES)] and chronic (using sub-convulsive dose of PTZ) models of seizure and seizure-induced oxidative stress and cognitive impairment in Wistar rats. CH exhibited dose-dependent protection against PTZ-induced seizures; all doses of CH produced significant increases in latency to myoclonic jerks in comparison to the PTZ-administered rats. Additionally, in the MES model, CH at doses of 30 mg/kg, 60 mg/kg, and 120 mg/kg demonstrated dose-dependent protection against THLE. Moreover, CH also attenuated seizure score caused by a subconvulsive dose of PTZ in PTZ-induced kindling. CH exerted maximum protection at 120 mg/kg against PTZ- and MES-induced seizures, as well as PTZ-induced kindling. CH had a protective effect in both the acute and chronic models, which suggests its potential use in both absence and generalised seizures. In addition, CH is also protective against epileptogenic processes, which had been revealed by the result of PTZ-induced kindling. It has been demonstrated that flavonoids possess beneficial effects against epilepsy by modulating the expression of γ-aminobutyric acid (GABA) and glutamate decarboxylase 65 and restoring the glutamate/GABA balance [ 27 , 28 ]. Moreover, flavonoids also exhibited anti-convulsant effects mediated by both an inhibition of NMDA receptors at the glycine-binding site and an agonistic activity on benzodiazepine-binding site at GABA A receptors [ 29 ]. It is well known that glutamatergic and GABAergic synaptic transmission, as well as excitatory amino acids such as glutamate, glycine, and NMDA, are involved in the initiation and propagation of seizures, thereby initiating epileptic activity [ 30 ]. Although the mechanism underlying the antiepileptic effect of CH has not been evaluated in this study, agonistic activity at the benzodiazepine-binding site or a decrease in synaptic glutamate or NMDA release may be involved. We performed EPM tests, MWM tests, and passive avoidance task tests to assess cognitive function in acute and chronic models of rats. It was observed that there was significant cognition impairment in experimental rat models of epilepsy in terms of increased retention transfer latency in the EPM test, increased escape latency, and decreased time spent in target quadrants in the MWM test and a decrease in the retention latency in the passive avoidance test [ 3 , 5 ] which was in accordance with earlier studies [ 3 , 5 ]. CH treatment improved the cognitive function caused by PTZ-, MES- and sub-convulsive dose of PTZ-induced seizure. In addition, CH improves cognition deficits caused by chronic cerebral hypoperfusion [ 31 ] and also caused by diabetes [ 32 ] in rats. It has been demonstrated that seizure activity increases oxidative stress and decreases antioxidant defence mechanisms in the brain [ 3 , 5 ]. Moreover, an imbalance between oxidant and antioxidant results in seizure progression and is responsible for cognitive deficit [ 33 ]. It has been reported in earlier studies that antioxidant treatment improves cognitive impairment by reducing brain oxidative stress in rat models of traumatic brain injury and Alzheimer's disease [ 3 , 34 , 35 ]. In the present study, CH ameliorated increased levels of TBARS and NO and decreased levels of reduced glutathione and SOD in both cortex and hippocampus caused by PTZ-, MES- and sub-convulsive dose of PTZ-induced seizure by virtue of its antioxidant activity [ 32 , 36 ], which might be responsible for improved cognitive function in this study. The role of acetylcholinesterase (AChE) in epilepsy is well understood, as it serves as a chemical mediator and an important regulator of cortical and hippocampal function. It has an intrinsic role in the seizure cascade [ 37 ]. Further, it has been reported that seizures decreased the AChE activity in the epileptic rat brain, which resulted in cognitive deficit in epileptic rats [ 5 , 38 ]. In the present study, PTZ-, MES-, and subconvulsive doses of PTZ-induced seizures caused a significant decrease in AChE activity in both the cortex and hippocampus compared to the normal control, which may be responsible for the development of cognitive impairment in epileptic rats. Treatment with CH significantly restored AChE activity in both the cortex and hippocampus, which was impaired by PTZ-, MES-, and subconvulsive dose of PTZ-induced seizures, and may be responsible for the reversal of cognitive functions. Our result thus demonstrates the anticholinergic effect of CH, which is in accordance with reports of previous studies [ 39 , 40 ]. Conclusion The present study demonstrates that treatment with CH attenuates PTZ-, MES-, and subconvulsive dose of PTZ-induced seizures, as well as seizure-induced oxidative stress and cognitive deficits, in rats. Our results suggest that CH mitigates oxidative stress induced by seizures by virtue of its free radical scavenging effects. In addition, the restoration of AChE activity may help CH restore the levels of acetylcholine in the brain, which in turn improves learning and memory. Further studies are necessary to gain a better understanding of the underlying molecular mechanisms responsible for the beneficial effects of CH in epilepsy. Declarations Declaration of competing interest There is no conflict of interest. Acknowledgements None Funding Sources This work was supported by the All-India Institute of Medical Sciences, New Delhi, India, through an Intramural Research Grant (IRG) to conduct this research. Authorship contribution statement S.K: Writing – original draft, Software, Data curation, Conceptualization. K.R: Project administration, Investigation, Conceptualization. P.P: Supervision, Software, Project administration, Conceptualization. S.K: Writing – review & editing, Conceptualization. K.R: Supervision,Funding acquisition. S.C: Writing – review & editing, Writing – original draft. 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1","display":"","copyAsset":false,"role":"figure","size":99527,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of Chrysin (CH) on seizure severity in the kindled rats. The mean seizure score in the PTZ group was the highest over the experiment; (n=6 per group). Values are presented as mean ± SEM.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-8407071/v1/fd640b7ef280f536afded61b.png"},{"id":99798130,"identity":"ae7ed328-fcdc-4cb9-a1c9-fcae1cd73cfa","added_by":"auto","created_at":"2026-01-08 13:47:21","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":103930,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of Chrysin (CH) on initial and retention transfer latency in the elevated plus maze test in the \u003cstrong\u003eA)\u003c/strong\u003e PTZ- seizure model \u003cstrong\u003eB)\u003c/strong\u003e MES- seizure model \u003cstrong\u003eC)\u003c/strong\u003e PTZ-induced kindling in rats; (n=6 per group). Values are presented as mean ± SEM. **P\u0026lt;0.01, ***P\u0026lt;0.001; a- as compared with the control; b- as compared with the PTZ group and MES group.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8407071/v1/a671f2a21c801fe330811a98.png"},{"id":99796462,"identity":"3f6594b8-2738-4f3c-ba58-c9cd17ac52d2","added_by":"auto","created_at":"2026-01-08 13:41:56","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":69799,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of Chrysin (CH) on escape latency in acquisition trials in probe trial in the Morris water maze test in the \u003cstrong\u003eA)\u003c/strong\u003e PTZ- seizure model \u003cstrong\u003eB)\u003c/strong\u003e MES- seizure model \u003cstrong\u003eC)\u003c/strong\u003e PTZ-induced kindling in rats; (n=6 per group). Values are presented as mean ±SEM. **P\u0026lt;0.01, ***P\u0026lt;0.001; a- as compared with the control; b- as compared with the PTZ group and MES group.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-8407071/v1/c47c8083cc850a47b11fabc2.png"},{"id":99797247,"identity":"cc43c8b0-18a9-4428-8aee-7f1765c26076","added_by":"auto","created_at":"2026-01-08 13:45:29","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":81079,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of Chrysin (CH) on time spent in the target quadrant in the Morris water maze test in the\u003cstrong\u003e A)\u003c/strong\u003e PTZ- seizure model \u003cstrong\u003eB)\u003c/strong\u003eMES- seizure model \u003cstrong\u003eC)\u003c/strong\u003e PTZ-induced kindling in rats; (n=6 per group). Values are presented as mean ± SEM. *P\u0026lt;0.05, **P\u0026lt;0.01, ***P\u0026lt;0.001; a- as compared with the control; b- as compared with the PTZ group and MES group.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-8407071/v1/ed8f93cb1fbf0f3cbe0f000f.png"},{"id":99796790,"identity":"d09391f2-0d07-4976-923f-b853bec7540c","added_by":"auto","created_at":"2026-01-08 13:43:40","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":199214,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of Chrysin (CH) on learning and memory in the passive avoidance test in the \u003cstrong\u003eA)\u003c/strong\u003e PTZ- seizure model \u003cstrong\u003eB)\u003c/strong\u003eMES- seizure model \u003cstrong\u003eC)\u003c/strong\u003e PTZ-induced kindling in rats; (n=6 per group). Values are presented as mean ± SEM. *P\u0026lt;0.05, **P\u0026lt;0.01, ***P\u0026lt;0.001; a- as compared with the control; b- as compared with the PTZ group and MES group.\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-8407071/v1/75386909631658844fac31a9.png"},{"id":99797970,"identity":"f0eb96d7-371e-469f-ba71-e24418189e71","added_by":"auto","created_at":"2026-01-08 13:47:01","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":157267,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of Chrysin (CH) on cortex and hippocampus thiobarbituric acid reactive substances (TBARS)\u003cem\u003e \u003c/em\u003ein the \u003cstrong\u003eA)\u003c/strong\u003e PTZ- seizure model \u003cstrong\u003eB)\u003c/strong\u003e MES- seizure model \u003cstrong\u003eC)\u003c/strong\u003e PTZ-induced kindling in rats; (n=6 per group). Values are presented as mean ± SEM. ***P\u0026lt;0.001; a- as compared with the control; b- as compared with the PTZ group and MES group.\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-8407071/v1/a6f34ba0f310b0ce7cd3fe12.png"},{"id":99797497,"identity":"935eb3bf-d582-404e-9ea7-2249500443b0","added_by":"auto","created_at":"2026-01-08 13:45:54","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":193751,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of Chrysin (CH) on cortex and hippocampus reduced glutathione (GSH)\u003cem\u003e \u003c/em\u003ein the \u003cstrong\u003eA)\u003c/strong\u003e PTZ- seizure model \u003cstrong\u003eB)\u003c/strong\u003e MES- seizure model \u003cstrong\u003eC)\u003c/strong\u003e PTZ-induced kindling in rats; (n=6 per group). Values are presented as mean ± SEM. ***P\u0026lt;0.001; a- as compared with the control; b- as compared with the PTZ group and MES group.\u003c/p\u003e","description":"","filename":"7.png","url":"https://assets-eu.researchsquare.com/files/rs-8407071/v1/0d51259e292e1937e86af809.png"},{"id":99721319,"identity":"165bde24-2e69-44d2-b245-ac15c5fa4773","added_by":"auto","created_at":"2026-01-07 15:25:38","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":194370,"visible":true,"origin":"","legend":"\u003cp\u003eEffect of Chrysin (CH) on cortex and hippocampus superoxide dismutase (SOD) in the \u003cstrong\u003eA)\u003c/strong\u003e PTZ- seizure model \u003cstrong\u003eB)\u003c/strong\u003eMES- seizure model \u003cstrong\u003eC)\u003c/strong\u003e PTZ-induced kindling in rats; (n=6 per group). Values are presented as mean ± SEM. ***P\u0026lt;0.001; a- as compared with the control; b- as compared with the PTZ group and MES group.\u003c/p\u003e","description":"","filename":"8.png","url":"https://assets-eu.researchsquare.com/files/rs-8407071/v1/8bbe761d670268c322ee5533.png"},{"id":100406357,"identity":"209b2877-e9a7-4fdd-80ed-d52a770daf54","added_by":"auto","created_at":"2026-01-16 13:01:00","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2249425,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8407071/v1/1fbb2b67-2e76-4082-a850-f48dc171c598.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Chrysin ameliorates seizures, seizure-induced oxidative stress and cognitive impairment in experimental models of epilepsy in rats","fulltext":[{"header":"Introduction","content":"\u003cp\u003eEpilepsy is one of the most prevalent and leading neurological disorders, with an incidence of about 1% of the population [1]. Moreover, it affects about 50 million people worldwide, and almost 2.4 million people are diagnosed with epilepsy every year [2]. The prevalence of epilepsy is higher in developing countries than in developed countries [3]. Many biological events contribute to the development and pathogenesis of epilepsy, including the inhibition of gamma-aminobutyric acid (GABA), a naturally occurring amino acid that functions as a neurotransmitter in the brain, as well as imbalanced inhibitory/excitatory transmissions and altered regulatory modulator systems [4]. \u0026nbsp;Cognitive impairment with abnormal behaviors in patients with epilepsy is observed, and a range of factors responsible for this, like anticonvulsant drugs, seizure type/frequency, structural lesions, and psychosocial stress [5]. In addition, oxidative stress (OS), which is associated with alterations in reactive oxygen species (ROS), reactive nitrogen species (RNS), and nitric oxide (NO) signaling pathways, whereby bioavailability of NO is decreased, and ROS and RNS production are increased. Further, oxidative damage is very common in the brain due to the abundance of mitochondria, high oxygen demand, poor repair capacity and high concentrations of polyunsaturated fatty acids, which play an important role in the neurological disorders, including epilepsy [6]. Moreover, there is involvement of the OS in the pathogenesis and progression of seizures, which adds to the cognitive disorders in patients with epilepsy [7].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIt has been demonstrated that there is change in cholinergic transmission in cognition impairment caused by seizure and assessment of cholinergic status in rat brain after seizures which can be done by evaluation of change in acetylcholinesterase activity too [8]. There is a significant number of antiepileptic drugs (AEDs) in the clinics for epileptic seizures, but these therapies target only symptoms of the diseases. They failed to adequately prevent seizure development or permanently halt the occurrence of seizures [9]. Moreover, AEDs is responsible for various side effects such as dizziness, tremor (shakiness), drowsiness, unsteadiness, blurred or double vision, bruising, and bleeding [10]. In additon,\u0026nbsp;more than 30% patients with epilepsy refractory to AEDs, which leads to a significant increase in the morbidity, mortality and impairment in quality of life in refractory patients of epilepsy [11]. There is an urgent need for newer and safer drugs to combat multi-factorial neurochemical abnormalities related to epileptic seizures.\u003c/p\u003e\n\u003cp\u003eFlavonoids, a subclass of polyphenols, are synthesised by plants as a secondary metabolite and are widely present in fruits, vegetables, and certain beverages. It has been demonstrated that flavonoids exhibit numerous beneficial effects in patients with epilepsy without causing any adverse side effects [10]. It has been demonstrated that the beneficial effects of flavonoids in neurological diseases may be associated with the modulation of GABA receptors, various voltage-gated ion channels, the regulation of antioxidant pathways, anti-inflammatory mediators, and the modulation of mitochondrial dysfunction and cytokine activity [10]. Chrysin (CH) (5,7-dihydroxy-2-phenyl-4H-chromen-4-one), a flavonoid, is a natural and most important bioactive constituent of honey, many fruits (like passion fruit (Passiflora sp.), kiwi, strawberries,\u0026nbsp;etc.),\u0026nbsp;vegetables and mushrooms [12, 13].\u0026nbsp;CH possesses an array of pharmacological properties such as antioxidative, neuroinflammatory, anti-depressant, anti-amyloidogenic, anticancer, and neuroprotective effects [12, 13].\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn view of these aforementioned facts, we designed this study to evaluate the effect of chrysin in pentylenetetrazol (PTZ) and maximal electroshock seizure (MES)-induced seizure, PTZ-induced kindling, seizures-induced oxidative stress and cognitive impairment in male Wistar rats.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eAnimals\u003c/h2\u003e \u003cp\u003e The study protocol was approved by the Institutional Animal Ethics Committee of the All-India Institute of Medical Sciences (AIIMS), New Delhi, India (843/IAEC/15). Male Wistar rats, weighing 150\u0026ndash;225 g, were obtained from the Central Animal Facility at AIIMS, New Delhi, India. They were maintained under standard laboratory conditions at a temperature of 25\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C with a natural light-dark cycle, and a standard dry rat pellet diet and tap water were provided \u003cem\u003ead libitum\u003c/em\u003e. The rats were acclimated to the laboratory environment for a week, and the animals were randomly assigned to the experimental groups, consisting of six rats per group, after 7 days of acclimation.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eDrugs and Chemicals\u003c/h3\u003e\n\u003cp\u003eChrysin, pentylenetetrazol, sodium valproate and all chemicals used for the experiment were of analytical grade and were purchased from Sigma-Aldrich Co., USA.\u003c/p\u003e\n\u003ch3\u003ePentylenetetrazol (PTZ)-induced seizures\u003c/h3\u003e\n\u003cp\u003ePTZ solution (60 mg/kg, once) was freshly prepared in normal saline and administered intraperitoneally (i.p.), 30 minutes and 60 minutes after the administration of sodium valproate (300 mg/kg; i.p.), chrysin (CH) (in doses of 30 mg/kg, 60 mg/kg and 120 mg/kg; once orally) respectively. The dose of PTZ (60 mg/kg, once) was standardised in our laboratory as the 100% convulsant dose with minimal mortality in rats [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. We recorded the latency to myoclonic jerks and the incidence of generalised tonic\u0026ndash;clonic seizures (GTCS) with loss of righting reflex. Rats were monitored for 30 minutes after administration of PTZ. We also evaluated the chronic effect of CH (in the doses of 30 mg/kg, 60 mg/kg and 120 mg/kg; once daily; orally) in a chronic epileptic seizure (kindling) rat model, which was induced by administration of sub-convulsive doses of PTZ (30 mg/kg; i.p.) on alternate day (48\u0026thinsp;\u0026plusmn;\u0026thinsp;1 h). PTZ was administered up to day 52 or until seizure stage 5 on two consecutive trials were achieved, whichever was earlier. Sodium valproate (300 mg/kg; i.p.) was also used as a standard. The rats were observed for 30 min after each PTZ administration for convulsive behaviour. Seizure activity was evaluated according to the method reported by Racine [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe latencies to myoclonic jerks and GTCS were recorded and were transformed into seizure score (S) [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e], which was calculated using the following formula:\u003cdiv class=\"BlockQuote\"\u003e\u003cp\u003eS\u0026thinsp;=\u0026thinsp;1- (control latency/drug seizure latency)\u003c/p\u003e\u003c/div\u003e\u003c/p\u003e \u003cp\u003eRats were considered kindled if they exhibited stage 5 of seizures on two consecutive trials. For control rats, S\u0026thinsp;=\u0026thinsp;0 and S\u0026thinsp;=\u0026thinsp;1 for rats who did not develop seizures. Rats were also observed for 24 h mortality, if any.\u003c/p\u003e\n\u003ch3\u003eMaximal electric shock (MES)-induced seizures\u003c/h3\u003e\n\u003cp\u003eRats were randomly divided into six groups. The first group served as vehicle control, the second group was the MES-induced seizure group, and the third group was treated with standard drug, i.e. sodium valproate (300 mg/kg; i.p.). The fourth, fifth, and sixth groups were administered CH 30 mg/kg, 60 mg/kg and 120 mg/kg, respectively. Electroconvulsions were produced in rats by a suprathreshold fixed-current sinus wave stimulus (70 mA, 0.2-second duration) delivered via ear-clip electrodes (Ugo Basile, Italy), 30 minutes and 60 minutes after administration of sodium valproate and CH, respectively. Rats were observed for the occurrence of tonic hind limb extension (THLE), i.e., the hind limbs of rats outstretched 180\u003csup\u003eo\u003c/sup\u003e to the plane of the body axis [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e\n\u003ch3\u003eCognitive function assessment\u003c/h3\u003e\n\u003cp\u003eThe behavioural parameters were performed before and 24 h after induction of seizures caused by PTZ or MES, and this was followed by CH administration. Morris water maze (MWM), elevated plus maze (EPM) and passive avoidance test (PAT) were conducted to assess any behavioural changes of the rats. Sixty minutes after CH administration, seizures were induced with either PTZ or MES. During the behavioural experiments, only one animal was tested at a time. Moreover, behavioural parameters were evaluated at baseline and after completion of the kindling procedure in the kindling experiment.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eElevated plus maze test\u003c/h2\u003e \u003cp\u003eThe elevated plus maze (EPM) test was performed to assess acquisition and retention memory processes as described earlier [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. The maze was placed 50 cm above the floor and consists of two closed arms and two open arms forming a cross, with a quadrangular centre. On day 1, the initial transfer latency was measured as follows: the rats were placed individually at the end of one open arm, facing away from the central platform, and the time it took to move from the open arm to either of the enclosed arms was recorded. Transfer latency was the time it took for the rats to move from the open arm to the enclosed arm, when all four of their legs crossed over to the enclosed arm. The rat was gently pushed onto its back into the enclosed arm, and a transfer latency of 60 seconds was assigned if the rat did not enter the enclosed arm within 60 seconds. After measuring the transfer latency, the rat was allowed to move freely in the plus maze, regardless of whether the arms were open or closed, for 10 seconds. The rat was then gently removed from the plus maze and returned to its home cage. Twenty-four hours later, the retention transfer latency test was performed in the same manner as in the acquisition trial. If the rat did not enter the enclosed arm within 60 seconds on the second trial, the transfer latency was assigned 60 seconds.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eMorris water maze\u003c/h3\u003e\n\u003cp\u003eMorris water maze was used to test the spatial memory of the rats. The testing procedure was used as described earlier [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Briefly, the water tank was divided into four equal quadrants (Q1, Q2, Q3 \u0026amp; Q4) and the platform was kept in the fourth quadrant. Rats were given four trials during four daily acquisition sessions. Every trial began by placing a rat in a water tank, facing the wall of the tank. Each of the four starting points was used once in a series of trials. The location of the platform was fixed for each trial. Further, the trial was terminated automatically as soon as the rat reached the platform or when 120 s had elapsed. The rat was allowed to continue on the platform for 5 seconds. Then it was removed from the platform, and a new trial was initiated. Rats that did not locate the platform within 120 seconds were gently placed on the platform and allowed to stay there for 5 seconds. After each trial, the rats were gently dried with a towel and returned to their home cage. On the fifth day, a spatial probe trial (60 seconds) was performed to assess the rat's spatial memory. On the 5th day, the same protocol was followed as described above; however, the platform had been removed from the tank. The path of each rat was analysed using the Any-maze video tracking system (Caterpillar Instrumentation Pvt. Ltd.). The latency to reach the platform and the time spent in the target quadrant (in which the platform was kept) were recorded.\u003c/p\u003e\n\u003ch3\u003e. One-trial passive avoidance task\u003c/h3\u003e\n\u003cp\u003eMemory retention deficit was evaluated by a step-through passive avoidance task [Ugo Basile, Italy (Model 7552)] as previously described [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Briefly, on the acquisition trial, each rat was placed in a lighted chamber. After 60 seconds of habituation, a guillotine door separating the lighted and dark chambers was opened, and the initial latency (IL) to enter the dark chamber was recorded. Rats exhibiting an initial latency time of more than 60 s were excluded from further experiments. Immediately after the rat entered the dark chamber, the guillotine door was closed, and an electric foot shock (75 V, 0.2 mA, 50 Hz) was delivered to the floor grids for 3 s. The rat was then removed from the dark chamber 5 seconds later and returned to its home cage. After 24 hours, retention latency (RL) time was measured in the same manner as during the acquisition trial, but foot shock was not delivered, and the latency time was recorded up to a maximum of 600 seconds.\u003c/p\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eBiochemical estimation\u003c/h2\u003e \u003cp\u003eAt the end of the experiments, all rats were sacrificed, and samples of brain tissue were collected, cleaned with ice-cold saline and stored at \u0026minus;\u0026thinsp;80\u0026deg;C until further analysis.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eTissue preparation\u003c/h2\u003e \u003cp\u003eBrain tissue samples were thawed, and the cortex and hippocampus from each cerebral hemisphere were separated for biochemical estimations. From separated parts (cortex and hippocampus), 10% w/v homogenates were prepared with ice-cold 0.1 M phosphate buffer (pH 7.4). Aliquots were prepared to determine lipid peroxidation, reduced glutathione, superoxide dismutase (SOD), acetylcholinesterase (AChE), nitric oxide, and protein estimation. The other half was preserved.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eMeasurement of lipid peroxidation\u003c/h2\u003e \u003cp\u003eTBARS (Thiobarbituric acid reactive substances), a measure of lipid peroxidation, was estimated as described earlier [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]. Briefly, acetic acid 375 \u0026micro;l (20%; pH 3.5), 375 \u0026micro;l thiobarbituric acid (0.8%), 50 \u0026micro;l sodium dodecyl sulfate (8.1%) and 20 \u0026micro;l water were added to 30 \u0026micro;l of processed brain tissue samples (cortex and hippocampus). The mixture was heated at 95\u0026deg;C for 60 minutes. The mixture was then cooled, and 1,000 \u0026micro;L of a 15:1 (v/v) n-butanol: pyridine solution was added. The mixture was then shaken vigorously. After centrifugation at 4000 rpm for 10 min, the organic layer was withdrawn, and absorbance was measured at 532 nm using a spectrophotometer.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eMeasurement of reduced glutathione (GSH)\u003c/h2\u003e \u003cp\u003eReduced glutathione was estimated as described by Ellman [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Briefly, the homogenate (cortex and hippocampus) was centrifuged with 5% trichloroacetic acid to precipitate the proteins. To 50 \u0026micro;l of this supernatant and 200 \u0026micro;l of 0.3 M phosphate buffer (pH 8.4), 25 \u0026micro;l of 5\u0026prime;5 dithiobis (2-nitrobenzoic acid) (DTNB) was added. The mixture was then vortexed, and the absorbance was read at 412 nm within 15 min.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eEstimation of nitrite\u003c/h2\u003e \u003cp\u003eThe accumulation of nitrite in the brain tissue (cortex and hippocampus) supernatant, an indicator of the production of nitric oxide (NO), which was determined as described by Green and his coworkers [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e], with Greiss reagent [0.1% N-(1-naphthyl) ethylenediamine dihydrochloride, 1% sulfanilamide, and 5% phosphoric acid]. Equal volumes of the supernatant (50 \u0026micro;L) and the Griess reagent (50 \u0026micro;L) were mixed, and the mixture was incubated at room temperature for 10 minutes. The absorbance was determined at 540 nm on an ELISA reader. The concentration of nitrite in the supernatant was determined from a sodium nitrite standard curve and was expressed as micromole per gram.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eSuperoxide dismutase (SOD)\u003c/h2\u003e \u003cp\u003eSuperoxide dismutase was estimated as described by Marklund and Marklund [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. Briefly, homogenized tissues with 10 times (w/v) 0.1 M sodium phosphate buffer (7.4). The reagent Tris-HCl-EDTA buffer (900 \u0026micro;l, pH 8.2) was added to 50 \u0026micro;l of the processed sample. Finally, at the time of reading, pyragallol (50 \u0026micro;L, 4 mM) was added, and optical observations were taken at 420 nm at different time intervals. The concentration of SOD was expressed as Units per Milligram of protein.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eEstimation of Cholinergic Status\u003c/h2\u003e \u003cp\u003eThe cholinergic markers, acetylcholinesterase, were estimated in the cortex and hippocampus of the brain tissue as described earlier by Ellman [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. Briefly, the rats' brains were removed over ice. The tissue was then homogenised in 0.1 M phosphate buffer and centrifuged at 10000 rpm for 30 min. at 4\u0026deg;C. 25 \u0026micro;l of this supernatant was incubated for 10 min with 650 \u0026micro;l buffer and 25 \u0026micro;l of DTNB (10 \u0026micro;M). Then, 25 \u0026micro;l of the freshly prepared acetylthiocholine iodide (75 mM) was used as substrate for the estimation of AChE. The absorbance was read at 412 nm for 3 min at 30, 60, 90, 120, 150 and 180 seconds.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eResults are represented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of the mean (SEM). One-way analysis of variance (ANOVA) followed by the Bonferroni multiple comparison test (SPSS-23 software) was done. A value of P\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered statistically significant.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eEffect of chrysin on Pentylenetetrazol (PTZ) induced seizures\u003c/h2\u003e \u003cp\u003ePTZ administration caused myoclonic jerks latency (54.6 second\u0026thinsp;\u0026plusmn;\u0026thinsp;6.1 second) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) and generalized tonic-colonic seizure latency (GTCS; 71.2 second\u0026thinsp;\u0026plusmn;\u0026thinsp;8.30 second) which was prevented by CH in dose dependent manner [myoclonic jerks (CH 30 mg/kg; 92.2 second\u0026thinsp;\u0026plusmn;\u0026thinsp;5.42 second), (CH 60 mg/kg; 127.7 second\u0026thinsp;\u0026plusmn;\u0026thinsp;6.7 second; P\u0026thinsp;\u0026lt;\u0026thinsp;0.01) and (CH 120 mg/kg; 136 second\u0026thinsp;\u0026plusmn;\u0026thinsp;7.9 second; P\u0026thinsp;\u0026lt;\u0026thinsp;0.001); GTCS (CH 30 mg/kg; 102 second\u0026thinsp;\u0026plusmn;\u0026thinsp;5.0 second), (CH 60 mg/kg; 150 second\u0026thinsp;\u0026plusmn;\u0026thinsp;9.4 second; P\u0026thinsp;\u0026lt;\u0026thinsp;0.01), (CH 120 mg/kg; 166 second\u0026thinsp;\u0026plusmn;\u0026thinsp;8.2 second; P\u0026thinsp;\u0026lt;\u0026thinsp;0.001] (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Moreover, the % age protection against PTZ-induce seizures was observed with CH 30 mg/kg, 60mg/kg and 120 mg/kg in dose dependent manner 66.7%, 83.3% and 100% respectively as compared to PTZ (0% protection). In addition, the %age protection against MES-induce seizures was also observed with CH 30 mg/kg, 60 mg/kg and 120 mg/kg in dose dependent manner 16.7%, 50% and 83.3% respectively as compared to MES (0% protection). Further, there was 100% protection against PTZ and MES in sodium valproate treated rats (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffect of the Chrysin (CH) on myoclonic jerk latency and occurrence of generalized tonic clonic seizures and tonic hindlimb extension in the pentylenetetrazol (PTZ) and maximal electric shock (MES)-induced seizure in rats (n\u0026thinsp;=\u0026thinsp;6 per group). Values are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM. ***P\u0026thinsp;\u0026lt;\u0026thinsp;0.001\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGroup\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eMyoclonic jerk latency (s) (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eLatency to onset of GTCS (s)\u003c/p\u003e \u003cp\u003e(mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e% Protection against\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eGeneralized tonic\u0026ndash;clonic seizures\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eTonic hind limb extension\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePTZ (60 mg/kg)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e54.6\u0026thinsp;\u0026plusmn;\u0026thinsp;6.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e71.2\u0026thinsp;\u0026plusmn;\u0026thinsp;8.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMaximal electric shock (70 mA)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e--- 0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSodium valproate (300 mg/kg)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e100 100\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCH30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e92.2\u0026thinsp;\u0026plusmn;\u0026thinsp;5.4\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e102\u0026thinsp;\u0026plusmn;\u0026thinsp;5.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e66.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCH60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e127.7\u0026thinsp;\u0026plusmn;\u0026thinsp;6.7\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e150\u0026thinsp;\u0026plusmn;\u0026thinsp;9.4\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e83.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCH120\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e135.9\u0026thinsp;\u0026plusmn;\u0026thinsp;7.9\u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e165.8\u0026thinsp;\u0026plusmn;\u0026thinsp;8.2 \u003csup\u003e***\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e83.3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eChronic administration of PTZ resulted in a gradual increase in seizure score over the course of the experiment, reaching a score of 4.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2. CH administration prevented the seizure severity in dose dose-dependent manner as compared to PTZ [(CH 120 mg/kg, 1.66\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2 vs 4.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2; P\u0026thinsp;\u0026lt;\u0026thinsp;0.001), (CH 60 mg/kg, 2.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4 vs 4.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2; P\u0026thinsp;\u0026lt;\u0026thinsp;0.01), (CH 30 mg/kg, 3.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5 vs 4.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2)]. Sodium valproate significantly protected the rats against PTZ-induced kindling (1 vs 4.33\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2, P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Therefore, the data from this study demonstrate that CH administration decreases the level of epileptic activity.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003eEffect of CH on Maximal electroshock seizures (MES) induced seizures\u003c/h2\u003e \u003cp\u003eThere was 100%, 50%, and 16.7% protection with CH 120 mg/kg, CH 60 mg/kg, and CH 30 mg/kg, respectively, against MES-induced THLE. None of the animals in the sodium valproate (VAL; 300 mg/kg)-treated group exhibited THLE \u003cb\u003e(\u003c/b\u003eTable\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e\u003cb\u003e)\u003c/b\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003eCognitive function\u003c/h2\u003e \u003cdiv id=\"Sec23\" class=\"Section3\"\u003e \u003ch2\u003eElevated plus maze (EPM)\u003c/h2\u003e \u003cp\u003eThere was no significant difference in the initial transfer latency among the groups, in all three models of epilepsy (PTZ, MES and PTZ-induced kindling). PTZ administration caused a significant increase in retention transfer latency (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) compared to the normal control (NC). Pretreatment with chrysin (30, 60 and 120 mg/kg) for 28 days significantly prevented the increase in retention transfer latency (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) in a dose-dependent manner as compared to PTZ (60 mg/kg) (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e2\u003c/span\u003eA), which was comparable to sodium valproate.\u003c/p\u003e \u003cp\u003eIn addition, MES caused a significant increase in the retention transfer latency (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) compared to the normal control (NC). Pretreatment with chrysin (30, 60, and 120 mg/kg) for 28 days significantly (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) decreased the retention transfer latency in a dose-dependent manner compared to MES \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e2\u003c/span\u003eB\u003cb\u003e)\u003c/b\u003e.\u003c/p\u003e \u003cp\u003eMoreover, in the chronic model, PTZ-induced kindling resulted in a significant increase in retention transfer latency (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) compared to the normal control (NC). Administration of chrysin (CH 30 mg/kg, 60 mg/kg, and 120 mg/kg; once daily) for 52 days significantly prevented the increase in retention transfer latency compared to PTZ (30 mg/kg), which was comparable to sodium valproate \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e2\u003c/span\u003eC\u003cb\u003e).\u003c/b\u003e\u003c/p\u003e \u003cp\u003eCH had no significant effect on retention transfer latency in all three epilepsy models compared to the control. Thus, CH improved memory retention, as indicated by significant decreases in retention transfer latency compared to seizure-induced rats.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003eMorris Water maze (MWM) test\u003c/h2\u003e \u003cp\u003eThere was a significant decrease in escape latency from days 1 to 4 in all groups, but no significant differences in escape latency were observed between groups on any of the four days in all three epilepsy models. There was a significant increase in escape latency (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and a decrease (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) in time spent in the target quadrant in all three models of epilepsy as compared to the normal control. In the PTZ (60 mg/kg) model, pretreatment with CH significantly (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001) decreased the escape latency and increased the time spent in the target quadrant in a dose-dependent manner as compared to PTZ (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e3\u003c/span\u003eA and \u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e4\u003c/span\u003eA). The effect of CH120 was comparable with that of sodium valproate \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e5\u003c/span\u003eA-B\u003cb\u003e)\u003c/b\u003e. However, CH \u003cem\u003eper se\u003c/em\u003e did not cause any significant change in either parameter in the MWM test compared to the control.\u003c/p\u003e \u003cp\u003eIn the MES model, chrysin pretreatment significantly decreased escape latency and increased time spent in the target quadrant in a dose-dependent manner as compared to the MES group. However, CH \u003cem\u003eper se\u003c/em\u003e did not cause any significant change in either parameter in the MWM test compared to the control \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e3\u003c/span\u003eB and \u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e4\u003c/span\u003eB\u003cb\u003e).\u003c/b\u003e\u003c/p\u003e \u003cp\u003eIn PTZ-induced kindling, CH (CH-60 mg/kg and 120 mg/kg, p.o, once daily) administration for 52 days significantly prevented the increase in escape latency, and CH (120 mg/kg) significantly prevented the decrease in the time spent in the target quadrant caused by PTZ. However, CH \u003cem\u003eper se\u003c/em\u003e did not cause any significant change in either parameter in the MWM test compared to the control \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e3\u003c/span\u003eC and \u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e4\u003c/span\u003eC\u003cb\u003e).\u003c/b\u003e\u003c/p\u003e \u003cdiv id=\"Sec25\" class=\"Section3\"\u003e \u003ch2\u003ePassive avoidance test\u003c/h2\u003e \u003cp\u003eThere was no significant difference in the initial latency among the different groups, in all three models of epilepsy (PTZ, MES and PTZ-induced kindling). PTZ (60 mg/kg) administration resulted in a significant decrease in retention latency compared to the normal control (P\u0026thinsp;\u0026lt;\u0026thinsp;0.001). Pretreatment with chrysin (30, 60 and 120 mg/kg) for 28 days significantly increased the retention latency in a dose-dependent manner as compared to PTZ (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e5\u003c/span\u003eA), which was comparable to valproate. CH \u003cem\u003eper se\u003c/em\u003e did not cause any significant change in retention latency as compared to control.\u003c/p\u003e \u003cp\u003eSubsequently, MES caused a significant decrease in the retention latency as compared to the normal control. Moreover, pretreatment with CH (30, 60, and 120 mg/kg) significantly prevented the decrease in retention latency in a dose-dependent manner caused by MES (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e5\u003c/span\u003eB). However, CH per se did not cause any significant change in retention latency compared to the control.\u003c/p\u003e \u003cp\u003eIn the chronic model of PTZ-induced kindling, PTZ administration (30 mg/kg) caused a significant decrease in retention latency compared to the normal control. CH (30 mg/kg, 60 mg/kg, and 120 mg/kg, p.o., once daily) administration caused a significant increase in retention latency compared to PTZ. However, chrysin \u003cem\u003eper se\u003c/em\u003e did not cause any significant change in retention latency as compared to control (Fig.\u0026nbsp;\u003cspan refid=\"Fig8\" class=\"InternalRef\"\u003e5\u003c/span\u003eC\u003cb\u003e)\u003c/b\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec26\" class=\"Section3\"\u003e \u003ch2\u003eBiochemical evaluation\u003c/h2\u003e \u003cdiv id=\"Sec27\" class=\"Section4\"\u003e \u003ch2\u003eThiobarbituric acid reactive substances (TBARS) levels\u003c/h2\u003e \u003cp\u003eA significant increase in TBARS levels was observed in brain tissues (cortex and hippocampus) in acute [PTZ (60 mg/kg) and MES] and chronic (PTZ-induced kindling) models of epilepsy, as compared to the normal control group. However, pretreatment with CH significantly attenuated the increased TBARS levels in a dose-dependent manner compared to PTZ, and the effect of CH was comparable to that of valproate (Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e6\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eIn the MES model, pretreatment with CH significantly prevented the dose-dependent increase in TBARS levels in the cortex and hippocampus caused by MES. The effect of CH on TBARS levels was comparable to that of valproate (Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e6\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003eIn the case of the chronic model (PTZ-induced kindling), administration of CH at a dose of 120 mg/kg significantly prevented the increase in TBARS levels in both the cortex and hippocampus caused by PTZ, which was comparable to that of valproate. Moreover, CH \u003cem\u003eper se\u003c/em\u003e did not cause any significant change in the levels of TBARS in all three epilepsy models compared to the control (Fig.\u0026nbsp;\u003cspan refid=\"Fig9\" class=\"InternalRef\"\u003e6\u003c/span\u003eC).\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec28\" class=\"Section2\"\u003e \u003ch2\u003eReduced glutathione levels\u003c/h2\u003e \u003cp\u003eThere was a significant decrease in the levels of reduced glutathione in the cortex and hippocampus in acute [PTZ (60 mg/kg) and MES] and chronic (PTZ-induced kindling) models of epilepsy as compared to the normal control group. However, pretreatment with three doses of CH (30 mg/kg, 60 mg/kg, and 120 mg/kg) significantly prevented the decrease in reduced glutathione levels in a dose-dependent manner in both the cortex and hippocampus, which was comparable to that of valproate (Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e7\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eIn the case of the MES model, CH pretreatment also significantly restored the levels of reduced glutathione in both the cortex and hippocampus in a dose-dependent manner, which were comparable to those of valproate (Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e7\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003eIn the case of PTZ-induced kindling, administration of CH at 60 mg/kg and 120 mg/kg significantly prevented the decrease in reduced glutathione levels in the cortex and hippocampus in a dose-dependent manner, as compared to PTZ, which was also comparable to the effect of valproate (Fig.\u0026nbsp;\u003cspan refid=\"Fig10\" class=\"InternalRef\"\u003e7\u003c/span\u003eC).\u003c/p\u003e \u003cp\u003eIn addition, CH \u003cem\u003eper se\u003c/em\u003e had no significant effect on reduced glutathione levels in both the cortex and hippocampus in all three epilepsy models, compared to the control.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec29\" class=\"Section2\"\u003e \u003ch2\u003eSuperoxide dismutase (SOD) levels\u003c/h2\u003e \u003cp\u003eIt was observed that the levels of SOD in the cortex and hippocampus significantly decreased in both the acute and chronic models of epilepsy as compared to the normal control. However, pretreatment with CH significantly restored the levels of SOD in the cortex and hippocampus in a dose-dependent manner, as compared to PTZ (60 mg/kg), which were comparable to those of valproate (Fig.\u0026nbsp;\u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e8\u003c/span\u003eA).\u003c/p\u003e \u003cp\u003eIn the case of the MES model, pretreatment of CH significantly prevented the decrease in the levels of SOD in the hippocampus, only in a dose-dependent manner, which was comparable with valproate. CH did not cause any significant change in the levels of SOD in the cortex as compared to MES (Fig.\u0026nbsp;\u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e8\u003c/span\u003eB).\u003c/p\u003e \u003cp\u003eIn the case of PTZ kindling, administration of CH significantly attenuated the decreased levels of SOD in the cortex and hippocampus in a dose-dependent manner, compared to PTZ, and this effect was also comparable to that of valproate. Moreover, CH \u003cem\u003eper se\u003c/em\u003e did not cause any significant change in three different models of epilepsy as compared to control (Fig.\u0026nbsp;\u003cspan refid=\"Fig11\" class=\"InternalRef\"\u003e8\u003c/span\u003eC).\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eNitric oxide (NO) levels\u003c/h3\u003e\n\u003cp\u003eThere was significant increase in the level of nitric oxide (NO) in all three model of epilepsy in cortex and hippocampus. Pretreatment of CH significantly prevented the increase in the levels of NO caused by PTZ (60 mg/kg) in a dose dependent manner in both cortex and hippocampus (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffect of CH on cortex and hippocampus nitric oxide (NO) levels in the \u003cb\u003eA)\u003c/b\u003e PTZ- seizure model \u003cb\u003eB)\u003c/b\u003e MES- seizure model \u003cb\u003eC)\u003c/b\u003e PTZ-induced kindling in rats; (n\u0026thinsp;=\u0026thinsp;6 per group). Values are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM. *P\u0026thinsp;\u0026lt;\u0026thinsp;0.05, ***P\u0026thinsp;\u0026lt;\u0026thinsp;0.001; a- as compared with the control; b- as compared with the PTZ group and MES group.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNO levels (\u0026micro;g/g of wet tissue) in cortex in acute PTZ\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003eNO levels (\u0026micro;g/g of wet tissue) in hippocampus in acute PTZ\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eNO levels (\u0026micro;g/g of wet tissue) in cortex in MES model\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNO levels (\u0026micro;g/g of wet tissue) in hippocampus in MES model\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eNO levels (\u0026micro;g/g of wet tissue) in cortex in chronic PTZ model\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNO levels (\u0026micro;g/g of wet tissue) in hippocampus in chronic PTZ model\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e48.0\u0026thinsp;\u0026plusmn;\u0026thinsp;2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e52.53\u0026thinsp;\u0026plusmn;\u0026thinsp;2.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e47.63\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e52.0\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e48.35\u0026thinsp;\u0026plusmn;\u0026thinsp;4.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e54.87\u0026thinsp;\u0026plusmn;\u0026thinsp;3.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePTZ (60 mg/kg)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e308.96\u0026thinsp;\u0026plusmn;\u0026thinsp;10.7\u003csup\u003e***a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e333.43\u0026thinsp;\u0026plusmn;\u0026thinsp;11.0\u003csup\u003e***a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMES (70 mA)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e166.80\u0026thinsp;\u0026plusmn;\u0026thinsp;6.3\u003csup\u003e***a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e180.94\u0026thinsp;\u0026plusmn;\u0026thinsp;15.5\u003csup\u003e***a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePTZ (Kindling)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e168.78\u0026thinsp;\u0026plusmn;\u0026thinsp;8.1\u003csup\u003e***a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e143.42\u0026thinsp;\u0026plusmn;\u0026thinsp;6.92\u003csup\u003e***a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSodium valproate (300 mg/kg)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e116.53\u0026thinsp;\u0026plusmn;\u0026thinsp;5.6\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e80.43 \u0026plusmn; 4.9\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e44.55\u0026thinsp;\u0026plusmn;\u0026thinsp;12.1\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e61.02\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e69.81\u0026thinsp;\u0026plusmn;\u0026thinsp;5.7\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e68.54\u0026thinsp;\u0026plusmn;\u0026thinsp;7.0\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCH30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e144.36\u0026thinsp;\u0026plusmn;\u0026thinsp;11.0\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e96.26\u0026thinsp;\u0026plusmn;\u0026thinsp;8.8\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e97.25\u0026thinsp;\u0026plusmn;\u0026thinsp;6.4\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e107.9\u0026thinsp;\u0026plusmn;\u0026thinsp;7.8\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e147.35\u0026thinsp;\u0026plusmn;\u0026thinsp;5.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e125.12\u0026thinsp;\u0026plusmn;\u0026thinsp;6.5\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCH60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e59.6\u0026thinsp;\u0026plusmn;\u0026thinsp;3.3\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e71.2\u0026thinsp;\u0026plusmn;\u0026thinsp;4.2\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e44.01\u0026thinsp;\u0026plusmn;\u0026thinsp;3.3\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e64.30\u0026thinsp;\u0026plusmn;\u0026thinsp;6.0\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e126.05\u0026thinsp;\u0026plusmn;\u0026thinsp;8.4\u003csup\u003e*b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e105.18\u0026thinsp;\u0026plusmn;\u0026thinsp;11.3\u003csup\u003e*b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCH120\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e32.80\u0026thinsp;\u0026plusmn;\u0026thinsp;4.8\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e39.63\u0026thinsp;\u0026plusmn;\u0026thinsp;4.8 \u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e29.25\u0026thinsp;\u0026plusmn;\u0026thinsp;3.9\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e59.32\u0026thinsp;\u0026plusmn;\u0026thinsp;5.8\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e65.74\u0026thinsp;\u0026plusmn;\u0026thinsp;6.3\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e77.96\u0026thinsp;\u0026plusmn;\u0026thinsp;7.6\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCH \u003cem\u003eper se\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e44.73\u0026thinsp;\u0026plusmn;\u0026thinsp;4.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e58.57\u0026thinsp;\u0026plusmn;\u0026thinsp;6.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e48.25\u0026thinsp;\u0026plusmn;\u0026thinsp;4.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e54.73\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50.29\u0026thinsp;\u0026plusmn;\u0026thinsp;6.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e49.23\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eIn case of MES model, CH pretreatment significantly prevented the increase in the levels of NO in cortex and hippocampus in dose dependent manner caused by MES (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn case of PTZ-induced kindling, administration of CH (120 mg/kg) significantly restored the levels of NO in cortex and hippocampus in dose dependent manner as compared to PTZ. CH at 30 mg/kg did not significantly decrease NO levels in cortex and hippocampus as compared to PTZ group (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e ).\u003c/p\u003e \u003cp\u003eThe effect of CH was comparable to that of valproate in all three epilepsy models. Moreover, chrysin \u003cem\u003eper se\u003c/em\u003e did not cause any significant change as compared to the control in all three different models of epilepsy.\u003c/p\u003e \u003cdiv id=\"Sec31\" class=\"Section2\"\u003e \u003ch2\u003eAcetylcholinesterase (AChE) levels\u003c/h2\u003e \u003cp\u003ePTZ (60 mg/kg), MES and PTZ-induced kindling caused significant decrease in the activity of AChE in both cortex and hippocampus as compared to normal control. Pretreatment with CH significantly increased the AChE activity in cortex and hippocampus in dose dependent manner as compared to PTZ. However, CH at the dose of 30 mg/kg and 60 mg/kg did not significantly prevent the decrease in AChE activity caused by PTZ (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eEffect of CH on cortex and hippocampus acetylcholinesterase (AChE) levels in the \u003cb\u003eA)\u003c/b\u003e PTZ- seizure model \u003cb\u003eB)\u003c/b\u003e MES- seizure model \u003cb\u003eC)\u003c/b\u003e PTZ-induced kindling in rats; (n\u0026thinsp;=\u0026thinsp;6 per group). Values are presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM. ***P\u0026thinsp;\u0026lt;\u0026thinsp;0.001; a- as compared with the control; b- as compared with the PTZ group and MES group.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAChE levels (Units) in cortex in acute PTZ\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAChE levels (Units) in hippocampus in in acute PTZ\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eAChE levels (Units) in cortex in in MES model\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAChE levels (Units) in hippocampus in in MES model\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAChE levels (Units) in cortex in chronic PTZ model\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eAChE levels (Units) in hippocampus in chronic PTZ model\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.81\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.84\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.62\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.69\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePTZ (60 mg/kg)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003csup\u003e***a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003e***a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMES (70 mA)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.45\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003e***a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003e***a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePTZ (Kindling)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e---\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.70\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003e***a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.64\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003e***a\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSodium valproate (300 mg/kg)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003e**b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.75\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCH30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.60\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.67\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.71\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCH60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.00\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.04\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003e*b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.44\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.69\u0026thinsp;\u0026plusmn;\u0026thinsp;0.06\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCH120\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.40\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003csup\u003e**b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.16\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e1.86\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.72\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e1.58\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003csup\u003e***b\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCH per se\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2.56\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.99\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.61\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.29\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e2.34\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.1\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eIn case of MES model, CH pretreatment significantly reversed the activity of AChE in both cortex and hippocampus in dose dependent manner as compared to MES challenges rats. CH at 30 mg/kg did not significantly reverse the decrease in AChE activity caused by MES (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn case of PTZ-induced kindling model, administration of CH significantly prevented the decrease in AChE levels in both cortex and hippocampus as compared to PTZ group. CH at 30 mg/kg did not significantly prevent the decrease in AChE activity caused by PTZ (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe effect of CH was comparable with valproate in all three different model of epilepsy. CH \u003cem\u003eper se\u003c/em\u003e had no significant effect on AChE activity as compared to control in all three different model of epilepsy.\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eEpilepsy is one of the common neurological disorders which still requires antiepileptic drugs (AEDs) with fewer side effects. A large number of antiepileptic drugs (AEDs) are available for the treatment of epilepsy, but treatment is still not satisfactory due to their side effects [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]. In addition, it has also been reported that epilepsy is responsible for cognitive impairment. Moreover, AEDs have also been shown to induce cognitive impairment in patients with epilepsy [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eCH is a natural flavonoid which occurs naturally in many plants, honey, and propolis. It possesses many biological activities and pharmacological effects, such as antioxidant, anti-inflammatory, anticancer, and antiviral activities [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. In addition, CH exhibits neuroprotective properties by inhibiting apoptosis [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eThe present study was conducted to evaluate the effect of CH in acute [induced chemically (PTZ; 60 mg/kg) and electrically (MES)] and chronic (using sub-convulsive dose of PTZ) models of seizure and seizure-induced oxidative stress and cognitive impairment in Wistar rats. CH exhibited dose-dependent protection against PTZ-induced seizures; all doses of CH produced significant increases in latency to myoclonic jerks in comparison to the PTZ-administered rats. Additionally, in the MES model, CH at doses of 30 mg/kg, 60 mg/kg, and 120 mg/kg demonstrated dose-dependent protection against THLE. Moreover, CH also attenuated seizure score caused by a subconvulsive dose of PTZ in PTZ-induced kindling. CH exerted maximum protection at 120 mg/kg against PTZ- and MES-induced seizures, as well as PTZ-induced kindling. CH had a protective effect in both the acute and chronic models, which suggests its potential use in both absence and generalised seizures. In addition, CH is also protective against epileptogenic processes, which had been revealed by the result of PTZ-induced kindling. It has been demonstrated that flavonoids possess beneficial effects against epilepsy by modulating the expression of γ-aminobutyric acid (GABA) and glutamate decarboxylase 65 and restoring the glutamate/GABA balance [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Moreover, flavonoids also exhibited anti-convulsant effects mediated by both an inhibition of NMDA receptors at the glycine-binding site and an agonistic activity on benzodiazepine-binding site at GABA\u003csub\u003eA\u003c/sub\u003e receptors [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. It is well known that glutamatergic and GABAergic synaptic transmission, as well as excitatory amino acids such as glutamate, glycine, and NMDA, are involved in the initiation and propagation of seizures, thereby initiating epileptic activity [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. Although the mechanism underlying the antiepileptic effect of CH has not been evaluated in this study, agonistic activity at the benzodiazepine-binding site or a decrease in synaptic glutamate or NMDA release may be involved.\u003c/p\u003e \u003cp\u003eWe performed EPM tests, MWM tests, and passive avoidance task tests to assess cognitive function in acute and chronic models of rats. It was observed that there was significant cognition impairment in experimental rat models of epilepsy in terms of increased retention transfer latency in the EPM test, increased escape latency, and decreased time spent in target quadrants in the MWM test and a decrease in the retention latency in the passive avoidance test [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e] which was in accordance with earlier studies [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. CH treatment improved the cognitive function caused by PTZ-, MES- and sub-convulsive dose of PTZ-induced seizure. In addition, CH improves cognition deficits caused by chronic cerebral hypoperfusion [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e] and also caused by diabetes [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e] in rats.\u003c/p\u003e \u003cp\u003eIt has been demonstrated that seizure activity increases oxidative stress and decreases antioxidant defence mechanisms in the brain [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Moreover, an imbalance between oxidant and antioxidant results in seizure progression and is responsible for cognitive deficit [\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. It has been reported in earlier studies that antioxidant treatment improves cognitive impairment by reducing brain oxidative stress in rat models of traumatic brain injury and Alzheimer's disease [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]. In the present study, CH ameliorated increased levels of TBARS and NO and decreased levels of reduced glutathione and SOD in both cortex and hippocampus caused by PTZ-, MES- and sub-convulsive dose of PTZ-induced seizure by virtue of its antioxidant activity [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e], which might be responsible for improved cognitive function in this study.\u003c/p\u003e \u003cp\u003eThe role of acetylcholinesterase (AChE) in epilepsy is well understood, as it serves as a chemical mediator and an important regulator of cortical and hippocampal function. It has an intrinsic role in the seizure cascade [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. Further, it has been reported that seizures decreased the AChE activity in the epileptic rat brain, which resulted in cognitive deficit in epileptic rats [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. In the present study, PTZ-, MES-, and subconvulsive doses of PTZ-induced seizures caused a significant decrease in AChE activity in both the cortex and hippocampus compared to the normal control, which may be responsible for the development of cognitive impairment in epileptic rats. Treatment with CH significantly restored AChE activity in both the cortex and hippocampus, which was impaired by PTZ-, MES-, and subconvulsive dose of PTZ-induced seizures, and may be responsible for the reversal of cognitive functions. Our result thus demonstrates the anticholinergic effect of CH, which is in accordance with reports of previous studies [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e].\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe present study demonstrates that treatment with CH attenuates PTZ-, MES-, and subconvulsive dose of PTZ-induced seizures, as well as seizure-induced oxidative stress and cognitive deficits, in rats. Our results suggest that CH mitigates oxidative stress induced by seizures by virtue of its free radical scavenging effects. In addition, the restoration of AChE activity may help CH restore the levels of acetylcholine in the brain, which in turn improves learning and memory. Further studies are necessary to gain a better understanding of the underlying molecular mechanisms responsible for the beneficial effects of CH in epilepsy.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eDeclaration of competing interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThere is no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNone\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding Sources\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by \u0026nbsp;the \u0026nbsp; All-India Institute of Medical Sciences, New Delhi, India, through an Intramural Research Grant (IRG) to conduct this research.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthorship contribution statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eS.K:\u0026nbsp;\u003c/strong\u003eWriting \u0026ndash; original draft, Software, Data curation, Conceptualization.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eK.R:\u0026nbsp;\u003c/strong\u003eProject administration, Investigation, Conceptualization.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eP.P:\u0026nbsp;\u003c/strong\u003eSupervision, Software, Project administration, Conceptualization.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eS.K:\u0026nbsp;\u003c/strong\u003eWriting \u0026ndash; review \u0026amp; editing, Conceptualization.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eK.R:\u0026nbsp;\u003c/strong\u003eSupervision,Funding acquisition.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eS.C:\u0026nbsp;\u003c/strong\u003eWriting \u0026ndash; review \u0026amp; editing, Writing \u0026ndash; original draft.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDisclosure of use of artificial intelligence (AI)-assistive or generative tools:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe AI tools or language models (LLMs) have not been utilised in the manuscript, except that software has been used for grammar corrections.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eBagheri S, Heydari A, Alinaghipour A, Salami M. 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Effect of melatonin on brain oxidative damage induced by traumatic brain injury in immature rats. Physiol Res 2005;54:631\u0026ndash;7.\u003c/li\u003e\n \u003cli\u003eReeta KH, Mehla J, Gupta YK. Curcumin ameliorates cognitive dysfunction and oxidative in phenobarbitone and carbamezepine administered rats. Eur J Pharmacol 2010;644:106\u0026ndash;12.\u003c/li\u003e\n \u003cli\u003eNaz S, Imran M, Rauf A, Orhan IE, Shariati MA, Iahtisham-Ul-Haq, IqraYasmin,Shahbaz M, Qaisrani TB, Shah ZA, Plygun S, Heydari M. Chrysin: Pharmacological and therapeutic properties. Life Sci. 2019;235:116797.\u003c/li\u003e\n \u003cli\u003eDiniz TC, Silva JC, de Lima-Saraiva SR, Ribeiro FP, Pacheco AG, de Freitas RM, Quintans-J\u0026uacute;nior LJ, Quintans Jde S, Mendes RL, Almeida JR. The role of flavonoids on oxidative stress in epilepsy. Oxid Med Cell Longev. 2015;2015:171756.\u003c/li\u003e\n \u003cli\u003ede Sales Santos IM, Feitosa CM, de Freitas RM. Pilocarpine-induced seizures produce alterations on choline acetyltransferase and acetylcholinesterase activities and deficit memory in rats. Cell Mol Neurobiol. 2010;30(4):569-75.\u003c/li\u003e\n \u003cli\u003eVedagiri A, Thangarajan S. Mitigating effect of chrysin loaded solid lipid nanoparticles against Amyloid \u0026beta;25-35 induced oxidative stress in rat hippocampal region: An efficient formulation approach for Alzheimer\u0026apos;s disease. Neuropeptides. 2016;58:111- 25.\u003c/li\u003e\n \u003cli\u003eTaslimi P, Kandemir FM, Demir Y, İlerit\u0026uuml;rk M, Temel Y, Caglayan C, Gul\u0026ccedil;in İ. The antidiabetic and anticholinergic effects of chrysin on cyclophosphamide-induced multiple organ toxicity in rats: Pharmacological evaluation of some metabolic enzyme activities. J Biochem Mol Toxicol. 2019:e22313.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Chrysin, Epilepsy, Pentylenetetrazol, Maximal electroshock seizure, Oxidative stress, Cognitive impairment","lastPublishedDoi":"10.21203/rs.3.rs-8407071/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8407071/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cb\u003ePurpose:\u003c/b\u003e\u003c/p\u003e \u003cp\u003eChrysin (CH), a naturally occurring flavonoid found in various traditional medicinal plants and bee products, has been reported to possess antioxidant and neuroprotective properties. These properties may offer therapeutic potential in epilepsy, a chronic neurological disorder frequently accompanied by cognitive impairment and oxidative stress. To evaluate the anticonvulsant, antioxidative, and cognitive-protective effects of chrysin in experimental models of epilepsy.\u003c/p\u003e\u003cp\u003e\u003cb\u003eMaterials and methods:\u003c/b\u003e\u003c/p\u003e \u003cp\u003eThe effects of CH were assessed using three established rodent models: pentylenetetrazol (PTZ)-induced seizures, maximal electroshock (MES)-induced seizures, and PTZ-induced kindling. Seizure severity, protection percentage, and progression of kindling were recorded. Cognitive performance was evaluated across all models, and biochemical assays were conducted to measure oxidative stress markers and acetylcholinesterase (AChE) activity.\u003c/p\u003e\u003cp\u003e\u003cb\u003eResults:\u003c/b\u003e\u003c/p\u003e \u003cp\u003eCH (120 mg/kg) demonstrated robust anticonvulsant activity, offering 83.3% protection against generalized tonic\u0026ndash;clonic seizures in the PTZ model and complete (100%) protection in the MES model. In PTZ-induced kindling, CH significantly attenuated epileptogenesis (mean seizure score\u0026thinsp;=\u0026thinsp;1.66). All three models exhibited marked cognitive deficits, which were significantly improved by CH treatment. CH also effectively restored oxidative stress parameters and reversed the seizure-induced reduction in AChE activity.\u003c/p\u003e\u003cp\u003e\u003cb\u003eConclusions:\u003c/b\u003e\u003c/p\u003e \u003cp\u003eChrysin exhibits potent anticonvulsant and neuroprotective effects in multiple seizure models, mitigating seizure severity, oxidative stress, and associated cognitive deficits. These findings support the ethnopharmacological relevance of chrysin-containing natural products and highlight CH as a promising phytochemical candidate for managing epilepsy and its neurocognitive complications.\u003c/p\u003e","manuscriptTitle":"Chrysin ameliorates seizures, seizure-induced oxidative stress and cognitive impairment in experimental models of epilepsy in rats","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-07 15:25:33","doi":"10.21203/rs.3.rs-8407071/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"a01a28ff-162e-4bde-ac0f-8c4a25a66bf3","owner":[],"postedDate":"January 7th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-01-08T13:44:09+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-07 15:25:33","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8407071","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8407071","identity":"rs-8407071","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}