Intervention Role Of The Structure Of Alpha-lipoic Acid On EAE Rat Central Nervous System Mitochondria And GSH, CAT

Objective: Multiple sclerosis (MS) is a chronic disease of the centralnervous system. Traditionally considered an inflammatory demyelinatingdisease, The main pathological features is that multiple central nervous systeminflammatory demyelination,glial cell proliferation and varying degrees ofaxonal damage. Recent evidence now points to axonal degeneration as crucialto the development of irreversible disability. The mechanisms of axonal andneuronal degeneration and neurologic disability in multiple sclerosis arepoorly understood. Oxidative injury to the mitochondrion classically heraldedas the mediators of demyelination and axonal injury by transection.Oxidative stress plays an important role in the pathogenesis of MS.Alpha-lipoic acid has attracted wide attention as a strong natural anti-oxidants.It mainly plays its antioxidant activity through scavenging free radicals,chelating metal ions, and regenerating other antioxidants. Alpha-lipoic acid asan antioxidant and immunity regulator that can protect and promote theremyelination in MS. the content of GSH in mitochondrial were examine toinvestigate the efect of mitoehondrial oxidative stress in the development ofCNS and protection of antioxidant treatment in EAE.Methods:1.Animal group: A total of96adult healthy female Wistar rats weredivided randomly into different groups: normal control group(NC,8rats),other rats were immunized by fresh guinea pig spinal cordhomogenate(GPSCH) and an equivalent complete Freund's adjuvant. Afterthey first paroxysm, divided randomly out6rats as disease group(DIS), othersdivided randomly into EAE group, dexamethasone(DM2mg·kg-1·d-1)group, alpha-lipoic acid (LA100mg·kg-1·d-1) group, and the3groups weredivided into Remission group(REM), relapse group(REL) two sub-groups according to the course of the disease, each sub-group6rats. No recurrence inDM group and; LA group, according to the separately mean days of remissionand relapse of EAE in preliminary experiments,the rats were divided into7days group after intervention (DM7d group; LA7d group)and14days groupafter intervention (DM14d group; LA14d group).2.Neural function assessment：Clinical signs of EAE were assessed themean of twice daily by two observations. Scores were assigned on the basis ofthe following symptoms:1, tail weakness;2, tail weakness plus limb asthenia;3, mild limb paralysis;4, severe limb paralysis;5, moribund.3.Histopathology： The brain and spinal cord tissues were got andembedded in paraffin. The sections were stained with HE staining,pathological changes were observed under light microscope. The ultrathinsections were observed under electron microscope for pathological changes ofmyelin sheath and axon.4. The CAT and GSH level in the tissues： The CAT and GSH levelactivity in the CNS tissues of rats was measured by CAT kid and GSH kid.Results:1.The relapse of disease in different groupComparison of the relapse of disease in three groups, the frequency ofEAE attacks in EAE group were higher than that the treatment groups DMgroup and LA group(P<0.05).2.Clinical profile of EAE in different groupNeurological deficits scores of EAE group were higher than those of DMgroup and LA group(P<0.05).3.Histopathology3.1HE staining: There were infiltration of inflammatory cells insidenerve tissue and perivascular cuffings in the junctional zone of gray and whitematter at the crest-time of the disease. Perivascular cells decreased or vanishedat paracmasis of the disease, and increased with conspicuous infiltration ofcells in white and grey matter at relapse of the disease.3.2Trichrome staining: Varying degrees axonal changes appeared in the other groups. The axon with inhomogeneous chromatosis was swollen andbeaded-discontinued; The myelin was porous and partly disaggregated.3.3Electron microscope: The mitochondria inside the axon were swollen,even with vacuolization. Mitochondrial swelling and dissolution of cristae alsohad disruption of the axonal cytoskeleton.4.The CAT level in the tissuesCAT level in the tissues of rats in LA and DM remitted group is higherthan other group(P<0.05). CAT level in the tissues of rats in LA and DMrelapsed is higher than the other guoup(P<0.05).5.The GSH level in the tissuesGSH level in the tissues of rats in LA and DM remitted group is higherthan other group(P<0.05). GSH level in the tissues of rats in LA and DMrelapsed is higher than the other guoup(P<0.05).Conclusions:1LA reduces the morbidity, and protects the rats from the severity of thedisease.2Through the HE staining, the modified trichrome staining and transmissionelectron microscope observation that the injury of myelin,axonal andmitochondrial in pathological changes are observed in the EAE rats.3Axonal injury and degeneration in pathological changes are observed inthe EAE rats.4Mitochondrial oxidative stress caused by may play an important role in thepathogenesis of EAE rat. Antioxidant a-LA could attenuate the oxidativedamage．...