Study On Anti-oxidative Stress Of Adenosine From Spores Powder Extraction Of Cordyceps Bassiana

Cordyceps bassiana is the traditional valuable medicinal material and food material in China,also known as cicada pupa,cicada velvet,cicada,scorpion,etc.,and belongs to the same genus Cordyceps as Cordyceps sinensis.The Cordyceps bassiana is rich in nucleoside substances and has various medicinal values.In nature,it has strict requirements on the growth environment.Therefore,it has not been extensively researched and developed,and foreign research on Cordyceps bassiana is scarce.In this paper,the use of high-purity Cordyceps bassiana spore powder to carry out in-depth research on the protective effects of its nucleoside active ingredients（especially adenosine）on the model animals Caenorhabditis elegans and PC-12 cells under oxidative stress conditions.The findings are as follows:1.Identification of bioactive components of C.bassiana spore powder and its effect on C.elegansThe nucleoside components in the extract of the bioactive components of C.bassiana spore powder（BC-CBSP）were identified,and the effect of the extract on C.elegans was investigated.The results show that different concentrations of BC-CBSP extract can prolong the life of nematodes.Within the experimental concentration range of 0.145mg/m L-14.5mg/m L,the higher the extract concentration,the longer the nematode life.The extract has a certain inhibitory effect on the reproductive capacity of the nematode.Among them,the low concentration extract delays the nematode spawning without affecting the total number of eggs,and the medium and high concentration extract slightly reduces the spawning amount.The high concentration of extracts inhibits the C.elegans’s ability to exercise.2.Establishment of C.elegans oxidative stress model and evaluation of protective effect of adenosine pretreatmentAn acute oxidative stress model was constructed by treating C.elegans with paraquat PQ.Through time-dose-mortality model（TDM）analysis,it was found that 150 m M PQ treatment made the nematode survival rate at 50%at 3 hours.On this basis,through the interaction of different concentrations of adenosine with the nematode acute oxidative stress model,it was found that medium and high concentrations of adenosine can significantly improve the survival rate of nematodes after PQ treatment,of which 3000μM adenosine has an acute oxidative stress Of nematodes have the highest protective effect.3.Transcriptome analysis of adenosine-protected C.elegans under oxidative stressTranscriptome sequencing analysis was performed on untreated nematodes,nematodes treated only with adenosine,nematodes under oxidative stress and under adenosine protection CK,AD,PQ,PQ＿AD.Based on the existing transcriptome information of Caenorhabditis elegans,the differentially expressed genes of each comparative group were screened,and the expression differences between CK vs PQ and PQ vs PQ＿AD were analyzed.Most of the differential genes in CK vs PQ and PQ vs PQ＿AD were expressed in opposite amounts.GO analysis of the common differential genes between the two groups revealed that several GO Term such as G-protein coupled receptor signaling pathway,Signaling,Extracellular region,Cell communication,Signal transduction and so on played a key role in the protection of nematodes by adenosine.Further analysis found that the family of neuropeptide genes including flp,nlp,npr,etc.has an important relationship with the protection of adenosine.4.Observation of the protective effect of adenosine under PC-12 cell modelThe protective effect of adenosine was further evaluated by the oxidative stress model of PC-12 cells.At a PQ concentration of 850 m M,the 24-hour survival rate of PC-12 cells is close to 50%.Adenosine pretreatment for 24 hours can protect PC-12 cells,and 30μM adenosine has the best protection effect.The research on the receptor mechanism of adenosine protection showed that A_2AR has no major role in protecting PQ injury.Mitochondrial membrane potentialΔΨwill decrease in PQ injured cells（P<0.001）,and cellular ROS will increase in injured cells（P<0.01）,but they will not recover in cells protected by adenosine（P>0.05）.