Cardarin effect on the formation of histopathological and behavioral abnormalities in the lithium-pilocarpine model of temporal lobe epilepsy in rats

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Abstract

Epilepsy is a severe neuropsychological disease accompanied by the development of spontaneous recurrent seizures (SRS) and associated behavioral disorders that are difficult to treat. In recent years, the neuroprotective properties of agonists of peroxisome proliferator-activated receptors (PPAR α, β/δ, γ), nuclear transcription factors involved in the regulation of lipid and carbohydrate metabolism, as well as inflammatory signaling pathways involved in the pathogenesis of epilepsy, have been actively investigated. The neuroprotective properties of PPARγ agonists have been repeatedly described in models of epilepsy; the effects of PPARβ/δ agonists in these models have not been sufficiently investigated. The aim of this work was to study the effects of administering the PPARβ/δ agonist cardarin on the formation of histopathological and behavioral abnormalities in the lithium-pilocarpine model of temporal lobe epilepsy (TLE). The lithium-pilocarpine model is one of the best experimental models of chronic temporal lobe epilepsy. In this study, epilepsy was induced by administration of pilocarpine to male Wistar rats at the age of 7 weeks one day after LiCl injection. Cardarin (2.5 mg/kg) was administered daily for 7 days after pilocarpine, with the first injection one day after pilocarpine injection. Behavioral testing was performed 2‒3 months after induction of the model in the following tests: Open Field, Resident Stranger, New Object Exploration, Y Maze Spontaneous Alternation and Morris Water Maze. Brain sampling for histological studies (assessment of neuronal death, Nissl staining) was performed after the end of behavioral experiments, 95 days after TLE induction. It was shown that untreated rats with TLE exhibited significant hippocampal neuron death and behavioral disorders: increased motor activity, anxiety, memory disorders, research and communicative behavior. Caradrin did not affect the survival rate of hippocampal neurons, but reduced the manifestation of almost all the above-mentioned behavioral disorders, except for hyperactivity. Thus, this study demonstrated the promising use of PPARβ/δ agonists to attenuate the development of behavioral disorders characteristic of epilepsy.

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M. R. Subkhankulov

Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences

Email: ZubarevaOE@mail.ru
Russian Federation, St. Petersburg

D. S. Sinyak

Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences

Email: ZubarevaOE@mail.ru
Russian Federation, St. Petersburg

V. A. Guk

Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences

Email: ZubarevaOE@mail.ru
Russian Federation, St. Petersburg

T. Yu. Postnikova

Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences

Email: ZubarevaOE@mail.ru
Russian Federation, St. Petersburg

A. I. Roginskaya

Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences

Email: ZubarevaOE@mail.ru
Russian Federation, St. Petersburg

O. E. Zubareva

Sechenov Institute of Evolutionary Physiology and Biochemistry of the Russian Academy of Sciences

Author for correspondence.
Email: ZubarevaOE@mail.ru
Russian Federation, St. Petersburg

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2. Fig.1. Experimental design.

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3. Fig.2. (a) – Kaplan-Meier survival curves. (b) – Dynamics of body weight. Cntr+Veh – control rats without treatment; Cntr+GW – control rats treated with cardarine; TLE+Veh – rats with temporal lobe epilepsy without treatment; TLE+GW – rats with temporal lobe epilepsy treated with cardarine. * – p < 0.05 difference between the Cntr+Veh and TLE+Veh groups; # – p < 0.05 difference between the Cntr+GW and TLE+GW groups, Shidak test.

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4. Fig.3. Neuronal death in the hippocampus of animals with temporal lobe epilepsy. (a) Examples of Nissl-stained hippocampal sections from control (Cntr+Veh), untreated (TLE+Veh) and treated (TLE+GW) rats. (b–c) – Number of neurons per 100 μm cell layer length in the hippocampal regions CA1 and CA3 in control (Cntr+Veh), untreated (TLE+Veh) and treated (TLE+GW) rats. **** – p < 0.0001, Shidak test. Each point corresponds to the indicator of an individual animal.

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5. Fig.4. Behavior of control and experimental rats in the Open Field test (a–g) and the Inspection of New Objects test (h, i). (a) – Examples of tracks in the Open Field; (b) – Total distance traveled in the Open Field; (c) – Locomotion time; (d) – Racks with emphasis, number of acts; (e) – Time in the central zone of the open field; (f) – Grooming time; (g) – Mink research time; (h) – Time to survey new objects; (i) – Number of contacts with new objects. Cntr+Veh – control rats without treatment; Cntr+GW – control rats treated with cardarine; TLE+Veh – rats with temporal lobe epilepsy without treatment; TLE+GW – rats with temporal lobe epilepsy treated with cardarine. * p < 0.05; ** p < 0.01; ***p < 0.001, ns – p > 0.05, Shidak or Dunn multiple comparison test. Each point corresponds to the indicator of an individual animal.

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6. Fig.5. Behavior of control and experimental rats in the social test. (a) – Communication time; (b) – Structure of communicative behavior; (c) – Time of aggression; (d) – % of rats with aggressive behavior (dark sector). Cntr+Veh – control rats without treatment; Cntr+GW – control rats treated with cardarine; TLE+Veh – rats with temporal lobe epilepsy without treatment; TLE+GW – rats with temporal lobe epilepsy treated with cardarine. * p < 0.05; ***p < 0.001, ns – p > 0.05, Shidak or Dunn multiple comparison test. Each point corresponds to the indicator of an individual animal.

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7. Fig.6. Behavior of control and experimental rats in the spontaneous alternation test in the Y-maze (a, b) and the Morris water maze (c–e). (a) – Alternation coefficient. (b) – Number of branches visited. (c) – Morris water maze training (distance traveled to find the platform over four training days). (d) – Test on the fifth day, time spent in the target area where the platform was previously located. (e) – Examples of tracks on the fifth day of testing. Cntr+Veh – control rats without treatment; Cntr+GW – control rats treated with cardarine; TLE+Veh – rats with temporal lobe epilepsy without treatment; TLE+GW – rats with temporal lobe epilepsy treated with cardarine. *, # , & p < 0.05; ** p<0.01; ns – p > 0.05, Shidak test (a, b) or Dunn test (c, d). (c): # – difference between the Cntr+Veh and TLE+Veh groups; & – between groups Cntr+GW and TLE+GW. Each dot represents a value for a specific animal.

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