| Objective:Through the production of epilepsy rat model, observe the effect of butylphthalideepileptic rats mitochondrial respiratory chain complexes Ⅱ, Ⅳ activity and the impact onthe electrical activity of cell populations in the hippocampal CA3region of mitochondrialultrastructure of the hippocampal CA3region, to explore the impact on the epilepsyhippocampal mitochondrial structure and function, or whether it can reduce the abnormalelectrical activity of the hippocampal CA3region through protection of mitochondrialfunction. This study investigates the relationship between epilepsy and mitochondrialfunction to seek epilepsy neurons protective measures, and provide a theoretical basis for theadjuvant treatment of epilepsy.Methods:Select80healthy adult female Wistar rats of8-10weeks, with60randomly divided intomodel group and20into control group. Establish the opentylenetetrazole kindling rat modelby intraperitoneal injection of freshly prepared1%pentylenetetrazo saline solution of35mg.kg-1.d-1. Refer to the Racine score standard, select them if three days in a row happenedwith Ⅳ-Ⅴ epilepsy row attack which was fully lit as the successful model. Since then1%pentylenetetrazol saline solution changed every3d administered the same dose to maintainthe kindling effect. Select the successful model after three weeks, and23were randomlydivided into butylphthalide group while24in epilepsy group. Butylphthalide group wasgiven butylphthalide solution80mg.kg-1.d-1, with the epileptic group given an equal volumeof peanut oil, while the intervention times are two weeks. Control group in the modelingperiod intraperitoneal injection of an equal volume of saline, with drug intervention periodgavage volume of normal saline. Observe the behavioral changes in each group, use thecolorimetric method to measure the activity of the hippocampal of rats in each group ofmitochondrial respiratory chain complexes Ⅱ, Ⅳ, and analyze with SPSS17.0statisticalsoftware; observe the transmission electron microscopy structure of hippocampal CA3region of mitochondrial using Advanced Micro Devices, and take radiography; collect and analyze the electrical activity of rat hippocampal CA3cell populations by adoptingstereotaxic apparatus and the BL-420biological functional system.Results:1. Observation of pentylenetetrazol-induced epileptic rats’ behavior: usually it happenedto be seizure2-3minutes after intraperitoneal injection in rats’ abdominal cavity. Firstly itmanifested with epileptic seizures, stiff tail, upturned, followed by fibrillation，clonus of thelimbs and loss of balance. The level of attack can be lighter to weighter step by step, or byleaps and bounds, with rapid emergence of the limbs twitch fell to the ground. Grand malepilepsy usually lasts2-3minutes and relieves itself, after which the animals appearrepetitive, rigid kind of local limb or muscle twitching. Usually it appears completeremission within30minutes, and the animal can do freely activities. Some animals have along incubation period, about5-6minutes after administration of seizures, very few seizuresin the administration of more than10minutes. A mold animals can be observed withspontaneous seizures, seizure frequency of1-3times a week, the attack to a lesser extent,multi-level Ⅰ-Ⅱ attack, occasionally Ⅳ attack, generally time of duration not more than1minute, with particular rat showing irritability.2. Rat hippocampal mitochondrial respiratory chain complex Ⅱ, Ⅳ activity: theactivity of control group, epilepsy group, butylphthalide group of complex Ⅱ activity（μmol DCPIP/min/mg） are respectively:1.8661±0.3491,1.4697±0.5580,1.7670±0.3073.Through independent samples t-test, the t value of control group and the epilepsy group is1.703, P value is0.111, P>0.05, the difference was not statistically significant; the t value ofbutylphthalide group and the epilepsy group is1.320, P value is0.208, P>0.05, thedifference was not statistically significant. The activity of control group, epilepsy group,butylphthalide group of complex Ⅳ activity （μmol Cytc/min/mg） are respectively:78.2967±7.6304,45.3296±11.6556,72.1153±15.1009. Through independent samples t-test, the tvalue of control group and the epilepsy group is6.693, P value is0.00019, P<0.001, thedifference was statistically significant; the t value of butylphthalide group and the epilepsygroup is3.972, P value is0.001, P<0.01, the difference was statistically significant.3. Ultrastructure of hippocampal CA3region: the control group neurons have rich nuclear matrix, rich organelles in the cytoplasm; neuropil processes pack tightly, and have alot of synapses. Mitochondrial membrane is integrity with a clear structure crest. Part ofepilepsy group neuron is swelling, chromatin in the nucleus is reduced, cytoplasm matrix iscavitated, organelles is reduced; some neurons showed dense dark cell-like, organelles in thecytoplasm are not easily distinguishable; neuropil protruding is cogitated, the number ofsynapses is reduced. Seen more mitochondria showed condensation or cavitation-like, severeinner and outer membrane rupture. Group butylphthalide has rich neuronal nuclearchromatin, rich organelles in the cytoplasm; rare dark cell neurons; protruding more withinthe neuropil, having a lot of synapses. Mitochondrial membrane is integrated with a clearridge, a small amount of dense mitochondrial structure.4. Electrical activity of hippocampal CA3cell populations: the control groupoccasionally occur too few, with less volatile explosive electrical activity（0.89times/min）;epilepsy group frequency hair explosive electrical activity（13.33times/min）, showing thatlarge amounts of spikes and the cluster of discharge; Butylphthalide group seen occasionallyhappened to very few and less volatile explosive electrical activity（3.00times/min）.Conclusion:1. Recurrent epilepsy can lead to hippocampal mitochondrial damage.2. Butylphthalide has a protective effect on epileptic rat hippocampal mitochondrialfunction, which significantly can improve the activity of the hippocampal of epileptic rats’mitochondrial respiratory chain complex Ⅳ; also reduce epilepsy hippocampalmitochondrial ultrastructural structural damage.3. Butylphthalide can reduce the electrical activity of epilepsy in rat hippocampal CA3area of the outbreak of the cell populations through protection of mitochondrial function,taken as the adjuvant treatment of epilepsy. |
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