Study Of The Disease-modifying Effects Of 1,6-diphosphate Fructose On Epilepsy Through Regulating The Astrocytes Metabolism

Objectives:Epilepsy is a complex neurological disease,characterized by spontaneous recurrent seizures.Traditional anti-epilepsy drugs reduce excessive excitability of neurons through targeting synaptic ion channels or neurotransmitter receptors,but they can only control the seizure behaviors but not improve the psychiatric comorbidities or slow disease progression.Recently,more and more evidences display that brain metabolic disorder plays an important role in epilepsy,and astrocytes are the key to maintain the metabolic homeostasis of neural circuits.1,6-diphosphate fructose(FBP)is an endogenous metabolite,which can modulate astrocyte metabolism.Our previous study found that FBP has a potential anti-seizure effect,and a significant cognitive protective effect on chronic epileptic animals.This study is to find the causal relationship between the metabolic regulation of FBP and its anti-seizure and neuroprotective effects,so as to clarify its function characteristics and possible mechanisms.Methods:(1)Anti-seizure and cognitive protective effects of FBP in the lithium-pilocarpine model of temporal lobe epilepsy(TLE):to compare seizure behaviors of valproate,levetiracetam and FBP groups during drug administration and after drug withdrawal;to evaluate the learning ability of each group through the Morris water maze experiment.(2)Compare the effects between FBP and another glycolysis inhibitors:to explore the effects of FBP treatment on glycolysis product lactate and neurotransmitters levels in the TLE model;to observe the effects of lactate shuttle blockers on lactate levels and neurotransmitter levels during acute epileptic seizures.(3)Effects of FBP on glucose metabolism and glutamate metabolism in astrocytes:to explore the effects of FBP treatment on the glucose and glutamate metabolic fluxs in different progresses in human astrocyte culture(HA)with isotopic labeled glutamate sodium and subsequent mass spectrometry detection.(4)To investigate the protective effect of FBP intervention on astrocytes and neurons degeneration in the TLE model through immunohistochemistry.Results:(1)In the TLE model,FBP showed faster,more stable anti-seizure effect than the positive control drugs,and especially only FBP could maintain the anti-seizure effect after drug withdraw.On the other hand,only FBP could protect or even improve the learning ability of chronic epileptic rats.(2)In the TLE model,FBP prevent the lactate levels from acute increasing under stimuli,and kept glutamate levels normal in interictal stage while retained its reasonable response to stimuli under the threshold.However,lactate shuttle blockers strongly atered lactate levels in intercellular space,and significantly reduced glutamate levels.(3)FBP switched glucose metabolism of HA cells from glycolysis to the tricarboxylic acid cycle,accelerated the clearance of glutamate through catabolism,conversion and biosynthesis,and greatly promoted the synthesis and release of glutathione.(4)FBP prevent astrocyte activation and gliosis,and protected neuron loss in chronic epileptic animals.Conclusions:FBP prevent seizures by maintaining lactate levels normal in the brain of chronic epileptic animals.On the other hand,FBP promotes the tricarboxylic acid cycle of astrocyte,enhances its glutamate scavenging and glutathione synthesis,thus reduces the excitatory toxicity and oxidative stress in pathological state.This study confirms that FBP is a potential anti-epilepsy drug with disease-modifying effecs and provides a noval strategy for anti-epilepsy treatment.