| Objective:Acute motor axonal neuropathy (AMAN) is a type of Gullian-Barre syndrome.There are many AMAN patients in HuaBei area,China.AMAN is marked by impairment of motor function and usually is serious,breathing difficulty in most patients.Perhaps who suffered from AMAN will develop tetraparesis in 24 to 48 hours,muscular atrophy very soon,and the incidence of illness and disability is high,prognosis being poor.In pathological aspect,there are severe to mild Wallerian-like degeneration,with rare demyeliantion or lymphocytic infiltration in anterior roots of spinal cords,sciatec nerves and plexus brachialis nerves in AMAN patients.But AMAN is different from acute inflammatory demyelinating polyneuropathy (AIDP).AIDP has its animal model which is generally acknowledged.We can induce experimental autoimmune neuropathy (EAN) in Lewis rats by sensitization with P2 protein from Bovine and pathological and clinical findings in EAN is similar to what in AIDP.The incident of EAN is high and stable .However,as for animal model of AMAN,there is no one like EAN.Though chickens developed flaccid paresis of limbs afterinoclulated with Campylobacter jejuni (Cj) in our laboratory,the incident is lower and not very stable. For this reason,we want to study a kind of means by which most of animals would develop paresis and there were Wallerian-like degeneration in pathological aspects.Gangliosides,being a set of acid glycolipids,which consist of lipid and saccharide,are the chief components of nerve cell membrane in mammal.In nervous system,GM1,GD1a,GD1b,GT1b,these four kinds of gangliosides,are highly enriched,especially GM1 being the most.However,out of nervous system,there are nearly none.High titers of serum anti-gangliosides antibodies,particularly anti-GM1 antibodies are found in 10 to 42% of patients with GBS.There are reports that sciatic nerve in rats injected with anti-GM1 antibodies positive serum from patients with multifocal motor neuropathy (MMN) developed conduction block and Igs deposition were found at the nodes of Ranvier.Then the sensitization of rabbits with GD1b did induce sensory neuropathy associated with anti-GD1b antibodies.These results indicated that gangliosides and anti-gangliosides antibodies were very important in pathogeny and mechanism of GBS.So we choice gangliosides to inoclulate white rabbits in order to establish animal model of AMAN. Material and Method:In the experiment,white rabbits used,the Bovine Brain Gangliosides (BBG) or GM1were injected subcutaneously to the back and intraperitoneally at 3 week intervals until limb weakness developed or 6 months had passed after the first inoculation.When rabbits developed limb weakness,they were killed at that day or 1 to 20 days after that day,on the basis of rabbits' condition.In brief, materials should be drawn as rabbits is dying naturally.Rabbits were slaughter by taken blood from the hearts .Then the left and the right sciatic nerves,the left and right plexus brachialis nerves,anterior and posterior roots were removed from rabbits.Nerve specimens were immersed in 33% glycerol for 12 hours,then in 66% glycerol for 12 hours,at last stored in 100% glycerol for teased fiber analysis.Besides,in the test,weekly blood samples were taken by ear vein puncture,then plasma samples were stored at -25℃ after centrifuge.The enzyme-linked immunosorbent assay (ELISA) was done for measuring anti-GM1 antibodies in plasma sample.At first,GM1 (250ng/100μl ethanol) were placed in individual wells of Ninety-six-well microtiter plates drying by airing.After being incubated with 1% bovine serum albumin (BSA) in PBS for 2 hours,plasma diluted 1:500 with 1% BSA in PBS was added to each well and each plasma sample was added to two wells,then the plate incubated at 4℃ for 24 hours.The wells were then washed with PBS for 5 times,and 100μl peroxidase-conjugated anti-rabbit γ-chain-specificantibody (IgG,1:2000 diluted) was added to each one.The plates were next incubated at room temperature for 2 hours,after which the plates were...
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