Anticonvulsive Effects of Chondroitin Sulfate on Pilocarpine and Pentylenetetrazole Induced Epileptogenesis in Mice
Abstract
:1. Introduction
2. Material and Method
2.1. Animals
2.2. Drugs and Chemicals
2.3. Anti-Convulsant Activity
- A. Induction of the Pentylenetetrazole Kindled Model of Epilepsy in Mice
- B. Induction of the Status Epilepticus Provoked Spontaneous Recurrent Seizure Activity in Mice
2.4. Estimation of Oxidative Stress
2.5. Estimation of Inflammatory Mediators
2.6. Histopathological Evaluation of Brain Tissue
2.7. Experimental Protocol
- A. Pentylenetetrazole-induced Kindled Seizure Severity Score Assessment Protocol (Figure 1)
- B. Pilocarpine-induced Spontaneous Recurrent Seizure Severity Score Assessment Protocol (Figure 2):
2.8. Molecular Docking Studies
2.9. Statistical Analysis
3. Results
3.1. Effect of Chondroitin Sulfate on PTZ-Induced Alteration in Kindling Severity Score in Mice
3.2. Effect of Chondroitin Sulfate on Pilocarpine-Induced Alteration in Spontaneous Recurrent Severity Score Mice
3.3. Effect of Chondroitin Sulfate on Pentylenetetrazole Mediated Oxidative Stress in Mice
3.4. Effect of Chondroitin Sulfate on Pilocarpine-Mediated Oxidative Stress in Mice
3.5. Effect of Chondroitin Sulfate on IL-1β, IL-6, NF-kB, and TNF-α Concentration in PTZ-Induced Kindling Seizures in Mice
3.6. Effect of Chondroitin Sulfate on IL-1β, IL-6, NF-kB, TNF-α, and Caspase-3 Concentration in Pilocarpine-Induced Spontaneous Seizures in Mice
3.7. Effect of Chondroitin Sulfate on PTZ-Ediated Caspase-3 Level in Mice
3.8. Effect of Chondroitin Sulfate on the Pilocarpine-Mediated Caspase-3 Level in Mice
3.9. Effect of Chondroitin Sulfate on PTZ- and Pilocarpine-Mediated Histopathological Changes in Mice Brains
3.10. Molecular Docking Studies
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
Sample Availability
References
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Groups | TBARs (nmol/mg of Protein) | Catalase (µM of H2O2 Oxidised/min/mg Protein) | Glutathione (µM of Glutathione/mg of Protein) | Superoxide Dismutase (SOD) (U/mg of protein) |
---|---|---|---|---|
Vehicle control group | 3.4 ± 0.99 | 39.73 ± 2.15 | 48.75 ± 2.76 | 35.8 ± 2.64 |
Pentylenetetrazole (PTZ) control | 22.1 ± 2.32 a | 14.26 ± 1.19 a | 17.01 ± 3.10 a | 12.95 ± 0.99 a |
Chondroitin sulfate (100 mg/kg) + PTZ treatment | 11.3 ± 1.57 b | 21.2 ± 1.75 b | 26.5 ±1.47 b | 20.93 ± 1.48 b |
Chondroitin sulfate (200 mg/kg) + PTZ treatment | 6.11 ± 0.57 b | 34.73 ± 1.60 b | 41.3 ± 1.53 b | 30.23 ± 1.24 b |
Valproic acid (110 mg/kg)+ PTZ treatment | 4.46 ± 0.52 b | 38.48 ± 2.09 b | 45.4 ± 3.93 b | 33.85 ± 3.46 b |
Groups | TBARs (nmol/mg of Protein) | Catalase (µM of H2O2 Oxidised/min/mg Protein) | Glutathione (µM of Glutathione/mg of Protein) | Superoxide Dismutase (SOD) (U/mg of Protein) |
---|---|---|---|---|
Vehicle control group | 3.4 ± 0.99 | 39.73 ± 2.15 | 48.75 ± 2.15 | 35.8 ± 2.64 |
Pilocarpine control | 24.76 ± 3.11 a | 11.36 ± 0.93 a | 15.38 ± 0.93 a | 10.86 ± 1.23 a |
Chondroitin sulfate (100 mg/kg) + pilocarpine treatment | 12.3 ± 1.78 b | 20.96 ± 1.53 b | 25.06 ± 1.53 b | 15.6 ± 0.81 b |
Chondroitin sulfate (200 mg/kg) + pilocarpine treatment | 6.38 ± 0.62 b | 34.06 ± 2.01 b | 39.96 ± 2.01 b | 29.56 ± 1.24 b |
Valproic acid (110 mg/kg) + pilocarpine treatment | 4.81 ± 0.71 b | 38.15 ± 2.67 b | 44.36 ± 2.67 b | 32.9 ± 2.02 b |
Groups | IL-1β (pg/mg of Tissue) | IL-6 (pg/mg of Tissue) | NF-kB (pg/mg of Tissue) | TNF-α (pg/mg of Tissue) | Caspase-3 (pg/mg of Tissue) |
---|---|---|---|---|---|
Vehicle control group | 47.25 ± 1.61 | 39.16 ± 2.37 | 36.83 ± 1.40 | 30.43 ± 1.19 | 0.71 ± 0.147 |
Pentylenetetrazole (PTZ) control | 155.86 ± 3.79 a | 106.83 ± 2.99 a | 83.46 ± 3.19 a | 74.8 ± 2.38 a | 8.3 ± 0.43 a |
Chondroitin sulfate (100 mg/kg) + PTZ treatment | 110.4 ± 4.59 b | 80.5 ± 3.83 b | 66.15 ± 2.53 b | 55.68 ± 3.24 b | 6.45 ± 0.92 b |
Chondroitin sulfate (200 mg/kg) + PTZ treatment | 62.1 ± 3.87 b | 54.3 ± 2.59 b | 48.13 ± 2.18 b | 44.5 ± 2.75 b | 4.51 ± 0.46 b |
Valproic acid (110 mg/kg) + PTZ treatment | 54.75 ± 2.16 b | 45.16 ± 1.45 b | 41.08 ± 1.58 b | 37.56 ± 1.35 b | 3.68 ± 0.33 b |
Groups | IL-1β (pg/mg of Tissue) | IL-6 (pg/mg of Tissue) | NF-kB (pg/mg of Tissue) | TNF-α (pg/mg of Tissue) | Caspase-3 (pg/mg of Tissue) |
---|---|---|---|---|---|
Vehicle control group | 47.25 ± 1.61 | 39.61± 2.37 | 36.88 ± 1.40 | 30.43± 1.19 | 0.71 ± 0.14 |
Pilocarpine control | 175.66 ± 4.36 a | 131.83± 4.73 a | 93.18± 1.92 a | 83.46± 3.83 a | 11.01 ± 0.90 a |
Chondroitin sulfate (100 mg/kg) + pilocarpine treatment | 120.66 ± 3.66 b | 93.51 ± 3.11 b | 68.48 ± 2.57 b | 58.55 ± 4.02 b | 6.6 ± 0.49 b |
Chondroitin sulfate (200 mg/kg) + pilocarpine treatment | 64.13 ± 3.99 b | 59.3 ± 2.79 b | 51.98 ± 2.45 b | 46.35 ± 3.52 b | 4.8 ± 0.52 b |
Valproic acid (110 mg/kg) + pilocarpine treatment | 56.2 ± 3.41 b | 44.9 ± 2.56 b | 43.35 ± 1.96 b | 39.31 ± 1.20 b | 3.9 ± 0.25 b |
Protein | Binding Interaction Energy | Type of Interaction | Bonding Amino Acids |
---|---|---|---|
TNF-alpha | + | H-bond | Tyr 151, Ser 60, Leu120, Gly121 |
van der Waals | Tyr59, Leu57, Gln61, Tyr119, Gly122 | ||
IL-1β | −16.50 | H-bond | Leu134, Phe133 |
van der Waals | Gly135, Trp120, Asp142 | ||
IL-6 | −18.68 | H-bond | Gln175, Arg30, Asp26, Arg179, Arg182 |
van der Waals | Leu33, Leu178 |
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Singh, S.; Singh, T.G.; Singh, M.; Najda, A.; Nurzyńska-Wierdak, R.; Almeer, R.; Kamel, M.; Abdel-Daim, M.M. Anticonvulsive Effects of Chondroitin Sulfate on Pilocarpine and Pentylenetetrazole Induced Epileptogenesis in Mice. Molecules 2021, 26, 6773. https://doi.org/10.3390/molecules26226773
Singh S, Singh TG, Singh M, Najda A, Nurzyńska-Wierdak R, Almeer R, Kamel M, Abdel-Daim MM. Anticonvulsive Effects of Chondroitin Sulfate on Pilocarpine and Pentylenetetrazole Induced Epileptogenesis in Mice. Molecules. 2021; 26(22):6773. https://doi.org/10.3390/molecules26226773
Chicago/Turabian StyleSingh, Shareen, Thakur Gurjeet Singh, Manjinder Singh, Agnieszka Najda, Renata Nurzyńska-Wierdak, Rafa Almeer, Mohamed Kamel, and Mohamed M. Abdel-Daim. 2021. "Anticonvulsive Effects of Chondroitin Sulfate on Pilocarpine and Pentylenetetrazole Induced Epileptogenesis in Mice" Molecules 26, no. 22: 6773. https://doi.org/10.3390/molecules26226773