Investigation Of The Neuroprotection And Mechanisms Of Corbrin Capsule In Amyotrophic Lateral Sclerosis Mouse Model

Amyotrophic lateral sclerosis(ALS)is a progression neurodegenerative disease,which is characterized by the selective deterioration of upper and lower motor neurons,leading to muscle paralysis,respiratory deficiency,and death.Though the pathophysiology of ALS are not fully understood,several pathogenic factors have been involved in,such as inflammation,protein misfold and aggregation,oxidative stress,endoplasmic reticulum(ER)stress.Microglia cells may have multiple functions in the CNS development,adulthood and ageing.Neurodegenerative diseases,for example ALS,lead to CNS microenvironment changes,which may prime microglia.The primed microglia might change form anti-inflammation and potential neuroprotective phenotype to pro-inflammation phenotype,leading to cell injury and death.It is now generally recognized that microglia can be polarized into M1 and M2 phenotypes.Research reports that at the early stage of ALS,microglia mainly polarized to M2 phenotype,with the massive expression of anti-inflammation cytokines.At the late phase of ALS,more M1 microglias were observed by increased expression of pro-inflammatory cytokines.Though riluzole and edaravone are improved by Food and Drug Administration of United States,their benefits are limited.In China,it has been reported that many traditional Chinese medicine might improve symptom and prolong the survival of ALS patients.Cordyceps sinensis(Dong chong xia cao)is one kind of traditional Chinese medicine,which has many chemical components with multiple pharmacological actions.For example,Cordycepin,one of the main bioactive components,was found have the potent anti-inflammation,anti-oxidative,anti-tumor properties,and neuroprotective function.Corbrin capsule(CC)was derived from Cordyceps sinensis,the composition of which are identical to that of wild Cordyceps sinensis mainly containing and thus can be used as a substitute for wild Cordyceps sinensis.In china,it has been reported that Corbrin capsule was effective for chronic obstructive pulmonary disease and renal recipients.In this study,we chose CC as the therapy drug and evaluated its neuroprotective effect in SOD1G93A mice,at the same time explored the potential mechanisms.Aims:Now,ALS still lacks of specific effective treatment drugs and methods.Corbrin capsule was derived from the traditional Chinese medicine Cordyceps sinensis(Dong chong xia cao).We used SOD1G93Amouse model of ALS to systematically observe the neuroprotective properties of CC and explore its potential mechanisms.Methods:The SOD1G93A mice were given CC(200mg per grain)per day by intragastric administration from 64days of the age to death(lg/kg:2 Corbrin capsules were dissolved in 4ml drinking water,0.01ml/gram by intragastric administration).The rotarod test was used to assess the day of disease onset and death.At the age of 120 days,part mice were sacrificed for its spinal cords and muscles for further study,for example immunofluorescence and RT-PCR.We immunofluorescence and Niss staining were performed to exam and count the motor neurons in the anterior horns.The hematoxylin-eosin and nicotinamide adenine dinucleotide hydrogen staining were performed to evaluate the function of gastrocnemius muscle.The integrated density of microglia and astrocyte positive staining was measured.The numbers of CD86+Iba-land Arg-1+Iba-1 positive cells were counted.Relative expressions of inflammatory factors were evaluated by RT-PCR.Result:Though CC did not delay the disease onset,it significantly prolonged lifepan and disease duration in SOD1G93A mice model of ALS.More motor neuron survived in CC treatment than that in drinking water treatment.And the integrated density of microglia and astrocyte positive staining shew no significant difference.In CC treatment group,the number of CD86 and Iba-1 positive cells was decreased,and the number of Arg-1 and Iba-1 positive cells was increased.The relative expressions of TNF-? and IL-6 were decreased significantly.And the relative expressions of IL-1? was decreased,but with no significant change.Conclusion:CC might extend the survival in SOD1G93Amice model of ALS,and promote microglia polarization from M1 phenotype to M2 phenotype.CC might be a potential candidate for the treatment for ALS.