Protective effects of adenosine kinase gene-modified mesenchymal stem cell transplantation on the seizures and hippocampus neuron damage of LiCl-pilocarpine induced temporal lobe epilepsy
Objective To explore the effects of adenosine kinase(ADK)-inhibited mouse bone marrow mesenchymal stem cells(MSCs)intracerebral transplantation on spontaneous recurrent seizures and hippocampus neuron in LiCl-pilocarpine epilepsy model.Methods Lentiviral vectors containing miRNA inhibiting ADK were used to produce ADK knockdown bone MSCs(ADK--MSCs).The adenosine secre-tion amount was analyzed.The ADK--MSCs were transplanted into the hippocampus of LiCl-pilocarpine mouse model.The mice in control group were injected by saline,those in TLE group were injected by pilo-carpine,and those in ADK--MSCs group were transplanted by ADK--MSCs before pilocarpine kindling.At 28 d after kindling,video monitoring EEG was used to observe seizures.Nissl staining was used to as-sess hippocampus survival neurons.Immunohistochemistry and Western blotting were used to evaluate ap-optosis in hippocampus.The survival ADK--MSCs were observed by fluorescence microscope at 45 d after kindling.Statistical differences between means of two groups were analyzed using Student's t test.Results The concentration of ADK in the supernatant of 1 × 105 ADK--MSCs was 12.1 ng/ml.The ADK--MSCs group had lower seizures frequency as(8.20±2.38)times per week,than(16.40±3.20)times per week in the TLE group(t=4.584,P＜0.01).Nissl staining showed that ADK--MSCs group had more survival neurons in CA1 region(91.00±6.51 vs.71.80±5.06,t=5.199,P＜0.001)and CA3 region(64.20±7.98 vs.45.75±7.63,t=3.511,P＜0.01)than TLE group.The immunohistochemical posi-tive cells of active cysteinyl aspartate-specific protease(Caspase)-3 were decreased in the ADK--MSCs group compared to the TLE group.Western blotting confirmed that the expression of active Caspase-3 in hippocampus was reduced in the ADK--MSCs group compared to the TLE group(n=3,t=4.302,P＜0.05).At 45 d after kindling,survival green fluorescent protein(GFP)positive transplanted ADK--MSCs were observed in the hippocampus.Conclusion Intracerebral transplantation of ADK-inhibited MSCs can suppress spontaneous recurrent seizures and neuronal damage in LiCl-pilocarpine-induced temporal lobe epilepsy model.
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